Sample records for sample preparation chip

  1. Improved Identification of Membrane Proteins by MALDI-TOF MS/MS Using Vacuum Sublimated Matrix Spots on an Ultraphobic Chip Surface

    PubMed Central

    Poetsch, Ansgar; Schlüsener, Daniela; Florizone, Christine; Eltis, Lindsay; Menzel, Christoph; Rögner, Matthias; Steinert, Kerstin; Roth, Udo

    2008-01-01

    Integral membrane proteins are notoriously difficult to identify and analyze by mass spectrometry because of their low abundance and limited number of trypsin cleavage sites. Our strategy to address this problem is based on a novel technology for MALDI-MS peptide sample preparation that increases the success rate of membrane protein identification by increasing the sensitivity of the MALDI-TOF system. For this, we used sample plates with predeposited matrix spots of CHCA crystals prepared by vacuum sublimation onto an extremely low wettable (ultraphobic) surface. In experiments using standard peptides, an up to 10-fold gain of sensitivity was found for on-chip preparations compared with classical dried-droplet preparations on a steel target. In order to assess the performance of the chips with membrane proteins, three model proteins (bacteriorhodopsin, subunit IV(a) of ATP synthase, and the cp47 subunit from photosystem II) were analyzed. To mimic realistic analysis conditions, purified proteins were separated by SDS-PAGE and digested with trypsin. The digest MALDI samples were prepared either by dried-droplet technique on steel plates using CHCA as matrix, or applied directly onto the matrix spots of the chip surface. Significantly higher signal-to-noise ratios were observed for all of the spectra resulting from on-chip preparations of different peptides. In a second series of experiments, the membrane proteome of Rhodococcus jostii RHA1 was investigated by AIEC/SDS-PAGE in combination with MALDI-TOF MS/MS. As in the first experiments, Coomassie-stained SDS-PAGE bands were digested and the two different preparation methods were compared. For preparations on the Mass·Spec·Turbo Chip, 43 of 60 proteins were identified, whereas only 30 proteins were reliably identified after classical sample preparation. Comparison of the obtained Mascot scores, which reflect the confidence level of the protein identifications, revealed that for 70% of the identified proteins, higher scores were obtained by on-chip sample preparation. Typically, this gain was a consequence of higher sequence coverage due to increased sensitivity. PMID:19137096

  2. Towards a “Sample-In, Answer-Out” Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System

    PubMed Central

    Gulliksen, Anja; Keegan, Helen; Martin, Cara; O'Leary, John; Solli, Lars A.; Falang, Inger Marie; Grønn, Petter; Karlgård, Aina; Mielnik, Michal M.; Johansen, Ib-Rune; Tofteberg, Terje R.; Baier, Tobias; Gransee, Rainer; Drese, Klaus; Hansen-Hagge, Thomas; Riegger, Lutz; Koltay, Peter; Zengerle, Roland; Karlsen, Frank; Ausen, Dag; Furuberg, Liv

    2012-01-01

    The paper presents the development of a “proof-of-principle” hands-free and self-contained diagnostic platform for detection of human papillomavirus (HPV) E6/E7 mRNA in clinical specimens. The automated platform performs chip-based sample preconcentration, nucleic acid extraction, amplification, and real-time fluorescent detection with minimal user interfacing. It consists of two modular prototypes, one for sample preparation and one for amplification and detection; however, a common interface is available to facilitate later integration into one single module. Nucleic acid extracts (n = 28) from cervical cytology specimens extracted on the sample preparation chip were tested using the PreTect HPV-Proofer and achieved an overall detection rate for HPV across all dilutions of 50%–85.7%. A subset of 6 clinical samples extracted on the sample preparation chip module was chosen for complete validation on the NASBA chip module. For 4 of the samples, a 100% amplification for HPV 16 or 33 was obtained at the 1 : 10 dilution for microfluidic channels that filled correctly. The modules of a “sample-in, answer-out” diagnostic platform have been demonstrated from clinical sample input through sample preparation, amplification and final detection. PMID:22235204

  3. An X-ray transparent microfluidic platform for screening of the phase behavior of lipidic mesophases

    PubMed Central

    Khvostichenko, Daria S.; Kondrashkina, Elena; Perry, Sarah L.; Pawate, Ashtamurthy S.; Brister, Keith

    2013-01-01

    Lipidic mesophases are a class of highly ordered soft materials that form when certain lipids are mixed with water. Understanding the relationship between the composition and the microstructure of mesophases is necessary for fundamental studies of self-assembly in amphiphilic systems and for applications, such as crystallization of membrane proteins. However, the laborious formulation protocol for highly viscous mesophases and the large amounts of material required for sample formulation are significant obstacles in such studies. Here we report a microfluidic platform that facilitates investigations of the phase behavior of mesophases by reducing sample consumption, and automating and parallelizing sample formulation. The mesophases were formulated on-chip using less than 40 nL of material per sample and their microstructure was analyzed in situ using small-angle X-ray scattering (SAXS). The 220 μm-thick X-ray compatible platform was comprised of thin polydimethylsiloxane (PDMS) layers sandwiched between cyclic olefin copolymer (COC) sheets. Uniform mesophases were prepared using an active on-chip mixing strategy coupled with periodic cooling of the sample to reduce the viscosity. We validated the platform by preparing and analyzing mesophases of lipid monoolein (MO) mixed with aqueous solutions of different concentrations of β-octylglucoside (βOG), a detergent frequently used in membrane protein crystallization. Four samples were prepared in parallel on chip, by first metering and automatically diluting βOG to obtain detergent solutions of different concentration, then metering MO, and finally mixing by actuation of pneumatic valves. Integration of detergent dilution and subsequent mixing significantly reduced the number of manual steps needed for sample preparation. Three different types of mesophases typical for monoolein were successfully identified in SAXS data from on-chip samples. Microstructural parameters of identical samples formulated in different chips showed excellent agreement. Phase behavior observed on-chip corresponded well with that of samples prepared via the traditional coupled-syringe method (“off-chip”) using 300-fold larger amount of material, further validating the utility of the microfluidic platform for on-chip characterization of mesophase behavior. PMID:23882463

  4. Lab-on-a-chip modules for detection of highly pathogenic bacteria: from sample preparation to detection

    NASA Astrophysics Data System (ADS)

    Julich, S.; Kopinč, R.; Hlawatsch, N.; Moche, C.; Lapanje, A.; Gärtner, C.; Tomaso, H.

    2014-05-01

    Lab-on-a-chip systems are innovative tools for the detection and identification of microbial pathogens in human and veterinary medicine. The major advantages are small sample volume and a compact design. Several fluidic modules have been developed to transform analytical procedures into miniaturized scale including sampling, sample preparation, target enrichment, and detection procedures. We present evaluation data for single modules that will be integrated in a chip system for the detection of pathogens. A microfluidic chip for purification of nucleic acids was established for cell lysis using magnetic beads. This assay was evaluated with spiked environmental aerosol and swab samples. Bacillus thuringiensis was used as simulant for Bacillus anthracis, which is closely related but non-pathogenic for humans. Stationary PCR and a flow-through PCR chip module were investigated for specific detection of six highly pathogenic bacteria. The conventional PCR assays could be transferred into miniaturized scale using the same temperature/time profile. We could demonstrate that the microfluidic chip modules are suitable for the respective purposes and are promising tools for the detection of bacterial pathogens. Future developments will focus on the integration of these separate modules to an entire lab-on-a-chip system.

  5. Protein Chips Compatible with MALDI Mass Spectrometry Prepared by Ambient Ion Landing.

    PubMed

    Pompach, Petr; Benada, Oldřich; Rosůlek, Michal; Darebná, Petra; Hausner, Jiří; Růžička, Viktor; Volný, Michael; Novák, Petr

    2016-09-06

    We present a technology that allows the preparation of matrix-assisted laser desorption/ionization (MALDI)-compatible protein chips by ambient ion landing of proteins and successive utilization of the resulting protein chips for the development of bioanalytical assays. These assays are based on the interaction between the immobilized protein and the sampled analyte directly on the protein chip and subsequent in situ analysis by MALDI mass spectrometry. The electrosprayed proteins are immobilized on dry metal and metal oxide surfaces, which are nonreactive under normal conditions. The ion landing of electrosprayed protein molecules is performed under atmospheric pressure by an automated ion landing apparatus that can manufacture protein chips with a predefined array of sample positions or any other geometry of choice. The protein chips prepared by this technique are fully compatible with MALDI ionization because the metal-based substrates are conductive and durable enough to be used directly as MALDI plates. Compared to other materials, the nonreactive surfaces show minimal nonspecific interactions with chemical species in the investigated sample and are thus an ideal substrate for selective protein chips. Three types of protein chips were used in this report to demonstrate the bioanalytical applications of ambient ion landing. The protein chips with immobilized proteolytic enzymes showed the usefulness for fast in situ peptide MALDI sequencing; the lectin-based protein chips showed the ability to enrich glycopeptides from complex mixtures with subsequent MALDI analysis, and the protein chips with immobilized antibodies were used for a novel immunoMALDI workflow that allowed the enrichment of antigens from the serum followed by highly specific MALDI detection.

  6. Multifunctional sample preparation kit and on-chip quantitative nucleic acid sequence-based amplification tests for microbial detection.

    PubMed

    Zhao, Xinyan; Dong, Tao

    2012-10-16

    This study reports a quantitative nucleic acid sequence-based amplification (Q-NASBA) microfluidic platform composed of a membrane-based sampling module, a sample preparation cassette, and a 24-channel Q-NASBA chip for environmental investigations on aquatic microorganisms. This low-cost and highly efficient sampling module, having seamless connection with the subsequent steps of sample preparation and quantitative detection, is designed for the collection of microbial communities from aquatic environments. Eight kinds of commercial membrane filters are relevantly analyzed using Saccharomyces cerevisiae, Escherichia coli, and Staphylococcus aureus as model microorganisms. After the microorganisms are concentrated on the membrane filters, the retentate can be easily conserved in a transport medium (TM) buffer and sent to a remote laboratory. A Q-NASBA-oriented sample preparation cassette is originally designed to extract DNA/RNA molecules directly from the captured cells on the membranes. Sequentially, the extract is analyzed within Q-NASBA chips that are compatible with common microplate readers in laboratories. Particularly, a novel analytical algorithmic method is developed for simple but robust on-chip Q-NASBA assays. The reported multifunctional microfluidic system could detect a few microorganisms quantitatively and simultaneously. Further research should be conducted to simplify and standardize ecological investigations on aquatic environments.

  7. FISH-in-CHIPS: A Microfluidic Platform for Molecular Typing of Cancer Cells.

    PubMed

    Perez-Toralla, Karla; Mottet, Guillaume; Tulukcuoglu-Guneri, Ezgi; Champ, Jérôme; Bidard, François-Clément; Pierga, Jean-Yves; Klijanienko, Jerzy; Draskovic, Irena; Malaquin, Laurent; Viovy, Jean-Louis; Descroix, Stéphanie

    2017-01-01

    Microfluidics offer powerful tools for the control, manipulation, and analysis of cells, in particular for the assessment of cell malignancy or the study of cell subpopulations. However, implementing complex biological protocols on chip remains a challenge. Sample preparation is often performed off chip using multiple manually performed steps, and protocols usually include different dehydration and drying steps that are not always compatible with a microfluidic format.Here, we report the implementation of a Fluorescence in situ Hybridization (FISH) protocol for the molecular typing of cancer cells in a simple and low-cost device. The geometry of the chip allows integrating the sample preparation steps to efficiently assess the genomic content of individual cells using a minute amount of sample. The FISH protocol can be fully automated, thus enabling its use in routine clinical practice.

  8. Nitrogen Cycle Evaluation (NiCE) Chip for the Simultaneous Analysis of Multiple N-Cycle Associated Genes.

    PubMed

    Oshiki, Mamoru; Segawa, Takahiro; Ishii, Satoshi

    2018-02-02

    Various microorganisms play key roles in the Nitrogen (N) cycle. Quantitative PCR (qPCR) and PCR-amplicon sequencing of the N cycle functional genes allow us to analyze the abundance and diversity of microbes responsible in the N transforming reactions in various environmental samples. However, analysis of multiple target genes can be cumbersome and expensive. PCR-independent analysis, such as metagenomics and metatranscriptomics, is useful but expensive especially when we analyze multiple samples and try to detect N cycle functional genes present at relatively low abundance. Here, we present the application of microfluidic qPCR chip technology to simultaneously quantify and prepare amplicon sequence libraries for multiple N cycle functional genes as well as taxon-specific 16S rRNA gene markers for many samples. This approach, named as N cycle evaluation (NiCE) chip, was evaluated by using DNA from pure and artificially mixed bacterial cultures and by comparing the results with those obtained by conventional qPCR and amplicon sequencing methods. Quantitative results obtained by the NiCE chip were comparable to those obtained by conventional qPCR. In addition, the NiCE chip was successfully applied to examine abundance and diversity of N cycle functional genes in wastewater samples. Although non-specific amplification was detected on the NiCE chip, this could be overcome by optimizing the primer sequences in the future. As the NiCE chip can provide high-throughput format to quantify and prepare sequence libraries for multiple N cycle functional genes, this tool should advance our ability to explore N cycling in various samples. Importance. We report a novel approach, namely Nitrogen Cycle Evaluation (NiCE) chip by using microfluidic qPCR chip technology. By sequencing the amplicons recovered from the NiCE chip, we can assess diversities of the N cycle functional genes. The NiCE chip technology is applicable to analyze the temporal dynamics of the N cycle gene transcriptions in wastewater treatment bioreactors. The NiCE chip can provide high-throughput format to quantify and prepare sequence libraries for multiple N cycle functional genes. While there is a room for future improvement, this tool should significantly advance our ability to explore the N cycle in various environmental samples. Copyright © 2018 American Society for Microbiology.

  9. Optofluidic analysis system for amplification-free, direct detection of Ebola infection

    NASA Astrophysics Data System (ADS)

    Cai, H.; Parks, J. W.; Wall, T. A.; Stott, M. A.; Stambaugh, A.; Alfson, K.; Griffiths, A.; Mathies, R. A.; Carrion, R.; Patterson, J. L.; Hawkins, A. R.; Schmidt, H.

    2015-09-01

    The massive outbreak of highly lethal Ebola hemorrhagic fever in West Africa illustrates the urgent need for diagnostic instruments that can identify and quantify infections rapidly, accurately, and with low complexity. Here, we report on-chip sample preparation, amplification-free detection and quantification of Ebola virus on clinical samples using hybrid optofluidic integration. Sample preparation and target preconcentration are implemented on a PDMS-based microfluidic chip (automaton), followed by single nucleic acid fluorescence detection in liquid-core optical waveguides on a silicon chip in under ten minutes. We demonstrate excellent specificity, a limit of detection of 0.2 pfu/mL and a dynamic range of thirteen orders of magnitude, far outperforming other amplification-free methods. This chip-scale approach and reduced complexity compared to gold standard RT-PCR methods is ideal for portable instruments that can provide immediate diagnosis and continued monitoring of infectious diseases at the point-of-care.

  10. Autonomous microfluidic sample preparation system for protein profile-based detection of aerosolized bacterial cells and spores.

    PubMed

    Stachowiak, Jeanne C; Shugard, Erin E; Mosier, Bruce P; Renzi, Ronald F; Caton, Pamela F; Ferko, Scott M; Van de Vreugde, James L; Yee, Daniel D; Haroldsen, Brent L; VanderNoot, Victoria A

    2007-08-01

    For domestic and military security, an autonomous system capable of continuously monitoring for airborne biothreat agents is necessary. At present, no system meets the requirements for size, speed, sensitivity, and selectivity to warn against and lead to the prevention of infection in field settings. We present a fully automated system for the detection of aerosolized bacterial biothreat agents such as Bacillus subtilis (surrogate for Bacillus anthracis) based on protein profiling by chip gel electrophoresis coupled with a microfluidic sample preparation system. Protein profiling has previously been demonstrated to differentiate between bacterial organisms. With the goal of reducing response time, multiple microfluidic component modules, including aerosol collection via a commercially available collector, concentration, thermochemical lysis, size exclusion chromatography, fluorescent labeling, and chip gel electrophoresis were integrated together to create an autonomous collection/sample preparation/analysis system. The cycle time for sample preparation was approximately 5 min, while total cycle time, including chip gel electrophoresis, was approximately 10 min. Sensitivity of the coupled system for the detection of B. subtilis spores was 16 agent-containing particles per liter of air, based on samples that were prepared to simulate those collected by wetted cyclone aerosol collector of approximately 80% efficiency operating for 7 min.

  11. Expression Profiling Smackdown: Human Transcriptome Array HTA 2.0 vs. RNA-Seq

    PubMed Central

    Palermo, Meghann; Driscoll, Heather; Tighe, Scott; Dragon, Julie; Bond, Jeff; Shukla, Arti; Vangala, Mahesh; Vincent, James; Hunter, Tim

    2014-01-01

    The advent of both microarray and massively parallel sequencing have revolutionized high-throughput analysis of the human transcriptome. Due to limitations in microarray technology, detecting and quantifying coding transcript isoforms, in addition to non-coding transcripts, has been challenging. As a result, RNA-Seq has been the preferred method for characterizing the full human transcriptome, until now. A new high-resolution array from Affymetrix, GeneChip Human Transcriptome Array 2.0 (HTA 2.0), has been designed to interrogate all transcript isoforms in the human transcriptome with >6 million probes targeting coding transcripts, exon-exon splice junctions, and non-coding transcripts. Here we compare expression results from GeneChip HTA 2.0 and RNA-Seq data using identical RNA extractions from three samples each of healthy human mesothelial cells in culture, LP9-C1, and healthy mesothelial cells treated with asbestos, LP9-A1. For GeneChip HTA 2.0 sample preparation, we chose to compare two target preparation methods, NuGEN Ovation Pico WTA V2 with the Encore Biotin Module versus Affymetrix's GeneChip WT PLUS with the WT Terminal Labeling Kit, on identical RNA extractions from both untreated and treated samples. These same RNA extractions were used for the RNA-Seq library preparation. All analyses were performed in Partek Genomics Suite 6.6. Expression profiles for control and asbestos-treated mesothelial cells prepared with NuGEN versus Affymetrix target preparation methods (GeneChip HTA 2.0) are compared to each other as well as to RNA-Seq results.

  12. Sample preparation and detection device for infectious agents

    DOEpatents

    Miles, Robin R.; Wang, Amy W.; Fuller, Christopher K.; Lemoff, Asuncion V.; Bettencourt, Kerry A.; Yu, June

    2003-06-10

    A sample preparation and analysis device which incorporates both immunoassays and PCR assays in one compact, field-portable microchip. The device provides new capabilities in fluid and particle control which allows the building of a fluidic chip with no moving parts, thus decreasing fabrication cost and increasing the robustness of the device. The device can operate in a true continuous (not batch) mode. The device incorporates magnetohydrodynamic (MHD) pumps to move the fluid through the system, acoustic mixing and fractionation, dielectropheretic (DEP) sample concentration and purification, and on-chip optical detection capabilities.

  13. Self-powered integrated microfluidic point-of-care low-cost enabling (SIMPLE) chip

    PubMed Central

    Yeh, Erh-Chia; Fu, Chi-Cheng; Hu, Lucy; Thakur, Rohan; Feng, Jeffrey; Lee, Luke P.

    2017-01-01

    Portable, low-cost, and quantitative nucleic acid detection is desirable for point-of-care diagnostics; however, current polymerase chain reaction testing often requires time-consuming multiple steps and costly equipment. We report an integrated microfluidic diagnostic device capable of on-site quantitative nucleic acid detection directly from the blood without separate sample preparation steps. First, we prepatterned the amplification initiator [magnesium acetate (MgOAc)] on the chip to enable digital nucleic acid amplification. Second, a simplified sample preparation step is demonstrated, where the plasma is separated autonomously into 224 microwells (100 nl per well) without any hemolysis. Furthermore, self-powered microfluidic pumping without any external pumps, controllers, or power sources is accomplished by an integrated vacuum battery on the chip. This simple chip allows rapid quantitative digital nucleic acid detection directly from human blood samples (10 to 105 copies of methicillin-resistant Staphylococcus aureus DNA per microliter, ~30 min, via isothermal recombinase polymerase amplification). These autonomous, portable, lab-on-chip technologies provide promising foundations for future low-cost molecular diagnostic assays. PMID:28345028

  14. Upgrade Recycling of Cast Iron Scrap Chips towards β-FeSi₂ Thermoelectric Materials.

    PubMed

    Laila, Assayidatul; Nanko, Makoto; Takeda, Masatoshi

    2014-09-04

    The upgrade recycling of cast-iron scrap chips towards β-FeSi₂ thermoelectric materials is proposed as an eco-friendly and cost-effective production process. By using scrap waste from the machining process of cast-iron components, the material cost to fabricate β-FeSi₂ is reduced and the industrial waste is recycled. In this study, β-FeSi₂ specimens obtained from cast iron scrap chips were prepared both in the undoped form and doped with Al and Co elements. The maximum figure of merit ( ZT ) indicated a thermoelectric performance of approximately 70% in p-type samples and nearly 90% in n-type samples compared to β-FeSi₂ prepared from pure Fe and other published studies. The use of cast iron scrap chips to produce β-FeSi₂ shows promise as an eco-friendly and cost-effective production process for thermoelectric materials.

  15. Development of low fat potato chips through microwave processing.

    PubMed

    Joshi, A; Rudra, S G; Sagar, V R; Raigond, P; Dutt, S; Singh, B; Singh, B P

    2016-08-01

    Since snacks high in fats are known to be a significant source of fat and energy intake, these have been put in high dietary restraint category. Therefore, an attempt was made to process potato chips through microwave processing without incorporation of any oil in potato chips. Microwave processing of potato chips was done using microwave power varying from 180 to 600 W using constant sample size. Among eleven different drying models, Parabolic model was found to be the best fit through non-linear regression analysis to illustrate drying kinetics of potato chips. The structural, textural and colour attributes of microwaved potato chips were similar to commercial fried potato chips. It was found that at 600 W after 2.5-3.0 min of processing, potato chips gained the fracturability and crispiness index as that of commercial fried chips. Microwave processing was found suitable for processing of potato chips with low fat content (~3.09 vs 35.5 % in commercial preparation) and with acceptable sensory scores (≥7.6 on 9.0 point on hedonic scale vs 8.0 of control preparation).

  16. Improved chip design for integrated solid-phase microextraction in on-line proteomic sample preparation.

    PubMed

    Bergkvist, Jonas; Ekström, Simon; Wallman, Lars; Löfgren, Mikael; Marko-Varga, György; Nilsson, Johan; Laurell, Thomas

    2002-04-01

    A recently introduced silicon microextraction chip (SMEC), used for on-line proteomic sample preparation, has proved to facilitate the process of protein identification by sample clean up and enrichment of peptides. It is demonstrated that a novel grid-SMEC design improves the operating characteristics for solid-phase microextraction, by reducing dispersion effects and thereby improving the sample preparation conditions. The structures investigated in this paper are treated both numerically and experimentally. The numerical approach is based on finite element analysis of the microfluidic flow in the microchip. The analysis is accomplished by use of the computational fluid dynamics-module FLOTRAN in the ANSYS software package. The modeling and analysis of the previously reported weir-SMEC design indicates some severe drawbacks, that can be reduced by changing the microextraction chip geometry to the grid-SMEC design. The overall analytical performance was thereby improved and also verified by experimental work. Matrix-assisted laser desorption/ionization mass spectra of model peptides extracted from both the weir-SMEC and the new grid-SMEC support the numerical analysis results. Further use of numerical modeling and analysis of the SMEC structures is also discussed and suggested in this work.

  17. Automated cellular sample preparation using a Centrifuge-on-a-Chip.

    PubMed

    Mach, Albert J; Kim, Jae Hyun; Arshi, Armin; Hur, Soojung Claire; Di Carlo, Dino

    2011-09-07

    The standard centrifuge is a laboratory instrument widely used by biologists and medical technicians for preparing cell samples. Efforts to automate the operations of concentration, cell separation, and solution exchange that a centrifuge performs in a simpler and smaller platform have had limited success. Here, we present a microfluidic chip that replicates the functions of a centrifuge without moving parts or external forces. The device operates using a purely fluid dynamic phenomenon in which cells selectively enter and are maintained in microscale vortices. Continuous and sequential operation allows enrichment of cancer cells from spiked blood samples at the mL min(-1) scale, followed by fluorescent labeling of intra- and extra-cellular antigens on the cells without the need for manual pipetting and washing steps. A versatile centrifuge-analogue may open opportunities in automated, low-cost and high-throughput sample preparation as an alternative to the standard benchtop centrifuge in standardized clinical diagnostics or resource poor settings.

  18. Microfluidic, marker-free isolation of circulating tumor cells from blood samples

    PubMed Central

    Karabacak, Nezihi Murat; Spuhler, Philipp S; Fachin, Fabio; Lim, Eugene J; Pai, Vincent; Ozkumur, Emre; Martel, Joseph M; Kojic, Nikola; Smith, Kyle; Chen, Pin-i; Yang, Jennifer; Hwang, Henry; Morgan, Bailey; Trautwein, Julie; Barber, Thomas A; Stott, Shannon L; Maheswaran, Shyamala; Kapur, Ravi; Haber, Daniel A; Toner, Mehmet

    2014-01-01

    The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 107 cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d. PMID:24577360

  19. Rapid Automated Sample Preparation for Biological Assays

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Shusteff, M

    Our technology utilizes acoustic, thermal, and electric fields to separate out contaminants such as debris or pollen from environmental samples, lyse open cells, and extract the DNA from the lysate. The objective of the project is to optimize the system described for a forensic sample, and demonstrate its performance for integration with downstream assay platforms (e.g. MIT-LL's ANDE). We intend to increase the quantity of DNA recovered from the sample beyond the current {approx}80% achieved using solid phase extraction methods. Task 1: Develop and test an acoustic filter for cell extraction. Task 2: Develop and test lysis chip. Task 3:more » Develop and test DNA extraction chip. All chips have been fabricated based on the designs laid out in last month's report.« less

  20. Comparison of contamination of femoral heads and pre-processed bone chips during hip revision arthroplasty.

    PubMed

    Mathijssen, N M C; Sturm, P D; Pilot, P; Bloem, R M; Buma, P; Petit, P L; Schreurs, B W

    2013-12-01

    With bone impaction grafting, cancellous bone chips made from allograft femoral heads are impacted in a bone defect, which introduces an additional source of infection. The potential benefit of the use of pre-processed bone chips was investigated by comparing the bacterial contamination of bone chips prepared intraoperatively with the bacterial contamination of pre-processed bone chips at different stages in the surgical procedure. To investigate baseline contamination of the bone grafts, specimens were collected during 88 procedures before actual use or preparation of the bone chips: in 44 procedures intraoperatively prepared chips were used (Group A) and in the other 44 procedures pre-processed bone chips were used (Group B). In 64 of these procedures (32 using locally prepared bone chips and 32 using pre-processed bone chips) specimens were also collected later in the procedure to investigate contamination after use and preparation of the bone chips. In total, 8 procedures had one or more positive specimen(s) (12.5 %). Contamination rates were not significantly different between bone chips prepared at the operating theatre and pre-processed bone chips. In conclusion, there was no difference in bacterial contamination between bone chips prepared from whole femoral heads in the operating room and pre-processed bone chips, and therefore, both types of bone allografts are comparable with respect to risk of infection.

  1. Chip-LC-MS for label-free profiling of human serum.

    PubMed

    Horvatovich, Peter; Govorukhina, Natalia I; Reijmers, Theo H; van der Zee, Ate G J; Suits, Frank; Bischoff, Rainer

    2007-12-01

    The discovery of biomarkers in easily accessible body fluids such as serum is one of the most challenging topics in proteomics requiring highly efficient separation and detection methodologies. Here, we present the application of a microfluidics-based LC-MS system (chip-LC-MS) to the label-free profiling of immunodepleted, trypsin-digested serum in comparison to conventional capillary LC-MS (cap-LC-MS). Both systems proved to have a repeatability of approximately 20% RSD for peak area, all sample preparation steps included, while repeatability of the LC-MS part by itself was less than 10% RSD for the chip-LC-MS system. Importantly, the chip-LC-MS system had a two times higher resolution in the LC dimension and resulted in a lower average charge state of the tryptic peptide ions generated in the ESI interface when compared to cap-LC-MS while requiring approximately 30 times less (~5 pmol) sample. In order to characterize both systems for their capability to find discriminating peptides in trypsin-digested serum samples, five out of ten individually prepared, identical sera were spiked with horse heart cytochrome c. A comprehensive data processing methodology was applied including 2-D smoothing, resolution reduction, peak picking, time alignment, and matching of the individual peak lists to create an aligned peak matrix amenable for statistical analysis. Statistical analysis by supervised classification and variable selection showed that both LC-MS systems could discriminate the two sample groups. However, the chip-LC-MS system allowed to assign 55% of the overall signal to selected peaks against 32% for the cap-LC-MS system.

  2. CAPACITANCE SENSING OF MOISTURE CONTENT IN BIO-FUEL MATERIALS: A RAPID AND NONDESTRUCTIVE METHOD FOR WOOD CHIPS

    USDA-ARS?s Scientific Manuscript database

    Moisture content of wood chips is an important factor to be known in their utilization as biomass material. Several moisture measuring instruments are available in the market, but for most of these instruments, some sort of sample preparation is needed that involves sizing, grinding and weighing. T...

  3. On chip preconcentration and fluorescence labeling of model proteins by use of monolithic columns: device fabrication, optimization, and automation.

    PubMed

    Yang, Rui; Pagaduan, Jayson V; Yu, Ming; Woolley, Adam T

    2015-01-01

    Microfluidic systems with monolithic columns have been developed for preconcentration and on-chip labeling of model proteins. Monoliths were prepared in microchannels by photopolymerization, and their properties were optimized by varying the composition and concentration of the monomers to improve flow and extraction. On-chip labeling of proteins was achieved by driving solutions through the monolith by use of voltage then incubating fluorescent dye with protein retained on the monolith. Subsequently, the labeled proteins were eluted, by applying voltages to reservoirs on the microdevice, and then detected, by monitoring laser-induced fluorescence. Monoliths prepared from octyl methacrylate combine the best protein retention with the possibility of separate elution of unattached fluorescent label with 50% acetonitrile. Finally, automated on-chip extraction and fluorescence labeling of a model protein were successfully demonstrated. This method involves facile sample pretreatment, and therefore has potential for production of integrated bioanalysis microchips.

  4. A fully sealed plastic chip for multiplex PCR and its application in bacteria identification.

    PubMed

    Xu, Youchun; Yan, He; Zhang, Yan; Jiang, Kewei; Lu, Ying; Ren, Yonghong; Wang, Hui; Wang, Shan; Xing, Wanli

    2015-07-07

    Multiplex PCR is an effective tool for simultaneous multiple target detection but is limited by the intrinsic interference and competition among primer pairs when it is performed in one reaction tube. Dividing a multiplex PCR into many single PCRs is a simple strategy to overcome this issue. Here, we constructed a plastic, easy-to-use, fully sealed multiplex PCR chip based on reversible centrifugation for the simultaneous detection of 63 target DNA sequences. The structure of the chip is quite simple, which contains sine-shaped infusing channels and a number of reaction chambers connecting to one side of these channels. Primer pairs for multiplex PCR were sequentially preloaded in the different reaction chambers, and the chip was enclosed with PCR-compatible adhesive tape. For usage, the PCR master mix containing a DNA template is pipetted into the infusing channels and centrifuged into the reaction chambers, leaving the infusing channels filled with air to avoid cross-contamination of the different chambers. Then, the chip is sealed and placed on a flat thermal cycler for PCR. Finally, amplification products can be detected in situ using a fluorescence scanner or recovered by reverse centrifugation for further analyses. Therefore, our chip possesses two functions: 1) it can be used for multi-target detection based on end-point in situ fluorescence detection; and 2) it can work as a sample preparation unit for analyses that need multiplex PCR such as hybridization and target sequencing. The performance of this chip was carefully examined and further illustrated in the identification of 8 pathogenic bacterial genomic DNA samples and 13 drug-resistance genes. Due to simplicity of its structure and operation, accuracy and generality, high-throughput capacity, and versatile functions (i.e., for in situ detection and sample preparation), our multiplex PCR chip has great potential in clinical diagnostics and nucleic acid-based point-of-care testing.

  5. Preparation of Low-Input and Ligation-Free ChIP-seq Libraries Using Template-Switching Technology.

    PubMed

    Bolduc, Nathalie; Lehman, Alisa P; Farmer, Andrew

    2016-10-10

    Chromatin immunoprecipitation (ChIP) followed by high-throughput sequencing (ChIP-seq) has become the gold standard for mapping of transcription factors and histone modifications throughout the genome. However, for ChIP experiments involving few cells or targeting low-abundance transcription factors, the small amount of DNA recovered makes ligation of adapters very challenging. In this unit, we describe a ChIP-seq workflow that can be applied to small cell numbers, including a robust single-tube and ligation-free method for preparation of sequencing libraries from sub-nanogram amounts of ChIP DNA. An example ChIP protocol is first presented, resulting in selective enrichment of DNA-binding proteins and cross-linked DNA fragments immobilized on beads via an antibody bridge. This is followed by a protocol for fast and easy cross-linking reversal and DNA recovery. Finally, we describe a fast, ligation-free library preparation protocol, featuring DNA SMART technology, resulting in samples ready for Illumina sequencing. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  6. Integrated printed circuit board device for cell lysis and nucleic acid extraction.

    PubMed

    Marshall, Lewis A; Wu, Liang Li; Babikian, Sarkis; Bachman, Mark; Santiago, Juan G

    2012-11-06

    Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 μm × 300 μm × 3.7 cm microfluidic channel connecting two 15 μL reservoirs. We demonstrated this device by extracting pathogenic nucleic acids from 1 μL dispensed volume of whole blood spiked with Plasmodium falciparum. We dispensed whole blood directly onto an on-chip reservoir, and the system's integrated heaters simultaneously lysed and mixed the sample. We used isotachophoresis to extract the nucleic acids into a secondary buffer via isotachophoresis. We analyzed the convective mixing action with micro particle image velocimetry (micro-PIV) and verified the purity and amount of extracted nucleic acids using off-chip quantitative polymerase chain reaction (PCR). We achieved a clinically relevant limit of detection of 500 parasites per microliter. The system has no moving parts, and the process is potentially compatible with a wide range of on-chip hybridization or amplification assays.

  7. Handling Heavenly Jewels - 35 Years of Antarctic Meteorite Processing at Johnson Space Center

    NASA Technical Reports Server (NTRS)

    Satterwhite, C. E.; McBridge, K. M.; Harrington, R.; Schwarz, C. M.

    2011-01-01

    The ANSMET program began in 1976, and since that time more than 18,000 meteorites have been processed in the Meteorite Processing Lab at Johnson Space Center in Houston, TX[1]. The meteorites are collected and returned to JSC on a freezer truck and remain frozen until they are initially processed. Initial Processing of Meteorites: Initial processing involves drying the meteorites in a nitrogen glove box for 24 to 48 hours, photographing, measuring, weighing and writing a description of the interior and exterior. The meteorite is broken and a representative sample is sent to the Smithsonian Institution for classification. Newsletter & Requests: Once initial processing has been complete and the meteorites have been classified, the information is published in the Antarctic Meteorite Newsletter[2,3]. The newsletter is published twice yearly and is sent electronically to researchers around the world and is also available on line. Researchers are asked to fill out a request form and submit it to the Meteorite Working Group secretary. All sample requests will be reviewed by either the meteorite curator or the Meteorite Working Group de-pending on the type of meteorite and the research being conducted. Processing for Sample Requests: In the meteorite processing lab, meteorite samples are prepared several different ways. Most samples are prepared as chips obtained by use of stainless steel chisels in a chipping bowl or rock splitter. In special situations where a researcher needs a slab the meteorite samples can be bandsawed in a dry nitrogen glove box with a diamond blade, no liquids are ever introduced into the cabinet. The last type of sample preparation is thin/thick sections. The meteorite thin section lab at JSC can prepare standard 30-micron thin sections, thick sections of variable thickness (100 to 200 microns), or demountable sections using superglue. Information for researchers: It is important that re-searchers fill the sample request form completely, in order to make sure the meteorite is processed correctly[4]. Re-searchers should list any special requirements on the form, i.e. packaging of samples (poly vs. stainless), thick sections and thickness needed, superglue needed, interior chips, exterior chips, fusion crust, contamination issues, all concerns should be listed so processing can be done accurately and any concerns the researcher has can be addressed be-fore the meteorites are broken.

  8. The MicroActive project: automatic detection of disease-related molecular cell activity

    NASA Astrophysics Data System (ADS)

    Furuberg, Liv; Mielnik, Michal; Johansen, Ib-Rune; Voitel, Jörg; Gulliksen, Anja; Solli, Lars; Karlsen, Frank; Bayer, Tobias; Schönfeld, Friedhelm; Drese, Klaus; Keegan, Helen; Martin, Cara; O'Leary, John; Riegger, Lutz; Koltay, Peter

    2007-05-01

    The aim of the MicroActive project is to develop an instrument for molecular diagnostics. The instrument will first be tested for patient screening for a group of viruses causing cervical cancer. Two disposable polymer chips with reagents stored on-chip will be inserted into the instrument for each patient sample. The first chip performs sample preparation of the epithelial cervical cells while mRNA amplification and fluorescent detection takes place in the second chip. More than 10 different virus markers will be analysed in one chip. We report results on sub-functions of the amplification chip. The sample is split into smaller droplets, and the droplets move in parallel channels containing different dried reagents for the different analyses. We report experimental results on parallel droplet movement control using one external pump only, combined with hydrophobic valves. Valve burst pressures are controlled by geometry. We show droplet control using valves with burst pressures between 800 and 4500 Pa. We also monitored the re-hydration times for two necessary dried reagents. After sample insertion, uniform concentration of the reagents in the droplet was reached after respectively 60 s and 10 min. These times are acceptable for successful amplification. Finally we have shown positive amplification of HPV type 16 using dried enzymes stored in micro chambers.

  9. Novel immunoassay formats for integrated microfluidic circuits: diffusion immunoassays (DIA)

    NASA Astrophysics Data System (ADS)

    Weigl, Bernhard H.; Hatch, Anson; Kamholz, Andrew E.; Yager, Paul

    2000-03-01

    Novel designs of integrated fluidic microchips allow separations, chemical reactions, and calibration-free analytical measurements to be performed directly in very small quantities of complex samples such as whole blood and contaminated environmental samples. This technology lends itself to applications such as clinical diagnostics, including tumor marker screening, and environmental sensing in remote locations. Lab-on-a-Chip based systems offer many *advantages over traditional analytical devices: They consume extremely low volumes of both samples and reagents. Each chip is inexpensive and small. The sampling-to-result time is extremely short. They perform all analytical functions, including sampling, sample pretreatment, separation, dilution, and mixing steps, chemical reactions, and detection in an integrated microfluidic circuit. Lab-on-a-Chip systems enable the design of small, portable, rugged, low-cost, easy to use, yet extremely versatile and capable diagnostic instruments. In addition, fluids flowing in microchannels exhibit unique characteristics ('microfluidics'), which allow the design of analytical devices and assay formats that would not function on a macroscale. Existing Lab-on-a-chip technologies work very well for highly predictable and homogeneous samples common in genetic testing and drug discovery processes. One of the biggest challenges for current Labs-on-a-chip, however, is to perform analysis in the presence of the complexity and heterogeneity of actual samples such as whole blood or contaminated environmental samples. Micronics has developed a variety of Lab-on-a-Chip assays that can overcome those shortcomings. We will now present various types of novel Lab- on-a-Chip-based immunoassays, including the so-called Diffusion Immunoassays (DIA) that are based on the competitive laminar diffusion of analyte molecules and tracer molecules into a region of the chip containing antibodies that target the analyte molecules. Advantages of this technique are a reduction in reagents, higher sensitivity, minimal preparation of complex samples such as blood, real-time calibration, and extremely rapid analysis.

  10. Technologies for autonomous integrated lab-on-chip systems for space missions

    NASA Astrophysics Data System (ADS)

    Nascetti, A.; Caputo, D.; Scipinotti, R.; de Cesare, G.

    2016-11-01

    Lab-on-chip devices are ideal candidates for use in space missions where experiment automation, system compactness, limited weight and low sample and reagent consumption are required. Currently, however, most microfluidic systems require external desktop instrumentation to operate and interrogate the chip, thus strongly limiting their use as stand-alone systems. In order to overcome the above-mentioned limitations our research group is currently working on the design and fabrication of "true" lab-on-chip systems that integrate in a single device all the analytical steps from the sample preparation to the detection without the need for bulky external components such as pumps, syringes, radiation sources or optical detection systems. Three critical points can be identified to achieve 'true' lab-on-chip devices: sample handling, analytical detection and signal transduction. For each critical point, feasible solutions are presented and evaluated. Proposed microfluidic actuation and control is based on electrowetting on dielectrics, autonomous capillary networks and active valves. Analytical detection based on highly specific chemiluminescent reactions is used to avoid external radiation sources. Finally, the integration on the same chip of thin film sensors based on hydrogenated amorphous silicon is discussed showing practical results achieved in different sensing tasks.

  11. Electrical Chips for Biological Point-of-Care Detection.

    PubMed

    Reddy, Bobby; Salm, Eric; Bashir, Rashid

    2016-07-11

    As the future of health care diagnostics moves toward more portable and personalized techniques, there is immense potential to harness the power of electrical signals for biological sensing and diagnostic applications at the point of care. Electrical biochips can be used to both manipulate and sense biological entities, as they can have several inherent advantages, including on-chip sample preparation, label-free detection, reduced cost and complexity, decreased sample volumes, increased portability, and large-scale multiplexing. The advantages of fully integrated electrical biochip platforms are particularly attractive for point-of-care systems. This review summarizes these electrical lab-on-a-chip technologies and highlights opportunities to accelerate the transition from academic publications to commercial success.

  12. Microfluidic-Mass Spectrometry Interfaces for Translational Proteomics.

    PubMed

    Pedde, R Daniel; Li, Huiyan; Borchers, Christoph H; Akbari, Mohsen

    2017-10-01

    Interfacing mass spectrometry (MS) with microfluidic chips (μchip-MS) holds considerable potential to transform a clinician's toolbox, providing translatable methods for the early detection, diagnosis, monitoring, and treatment of noncommunicable diseases by streamlining and integrating laborious sample preparation workflows on high-throughput, user-friendly platforms. Overcoming the limitations of competitive immunoassays - currently the gold standard in clinical proteomics - μchip-MS can provide unprecedented access to complex proteomic assays having high sensitivity and specificity, but without the labor, costs, and complexities associated with conventional MS sample processing. This review surveys recent μchip-MS systems for clinical applications and examines their emerging role in streamlining the development and translation of MS-based proteomic assays by alleviating many of the challenges that currently inhibit widespread clinical adoption. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  13. Special-purpose computing for dense stellar systems

    NASA Astrophysics Data System (ADS)

    Makino, Junichiro

    2007-08-01

    I'll describe the current status of the GRAPE-DR project. The GRAPE-DR is the next-generation hardware for N-body simulation. Unlike the previous GRAPE hardwares, it is programmable SIMD machine with a large number of simple processors integrated into a single chip. The GRAPE-DR chip consists of 512 simple processors and operates at the clock speed of 500 MHz, delivering the theoretical peak speed of 512/226 Gflops (single/double precision). As of August 2006, the first prototype board with the sample chip successfully passed the test we prepared. The full GRAPE-DR system will consist of 4096 chips, reaching the theoretical peak speed of 2 Pflops.

  14. Preparation of a molecularly imprinted sensor based on quartz crystal microbalance for specific recognition of sialic acid in human urine.

    PubMed

    Qiu, Xiuzhen; Xu, Xian-Yan; Chen, Xuncai; Wu, Yiyong; Guo, Huishi

    2018-05-08

    A novel molecularly imprinted quartz crystal microbalance (QCM) sensor was successfully prepared for selective determination of sialic acid (SA) in human urine samples. To obtain the QCM sensor, we first modified the gold surface of the QCM chip by self-assembling of allylmercaptane to introduce polymerizable double bonds on the chip surface. Then, SA molecularly imprinted polymer (MIP) nanofilm was attached to the modified QCM chip surface. For comparison, we have also characterized the nonmodified and improved surfaces of the QCM sensor by using atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectroscopy. We then tested the selectivity and detection limit of the imprinted QCM sensor via a series of adsorption experiments. The results show a linear response in the range of 0.025-0.50 μmol L -1 for sialic acid. Moreover, the limit of detection (LOD) of the prepared imprinted QCM sensor was found to be 1.0 nmol L -1 for sialic acid, and high recovery values range from 87.6 to 108.5% with RSD < 8.7 (n = 5) for the spiked urine sample obtained. Overall, this work presents how a novel QCM sensor was developed and used to detect sialic acid in human urine samples. Graphical abstract Specific recognition of sialic acid by the MIP-QCM sensor system.

  15. A hybrid approach to device integration on a genetic analysis platform

    NASA Astrophysics Data System (ADS)

    Brennan, Des; Jary, Dorothee; Kurg, Ants; Berik, Evgeny; Justice, John; Aherne, Margaret; Macek, Milan; Galvin, Paul

    2012-10-01

    Point-of-care (POC) systems require significant component integration to implement biochemical protocols associated with molecular diagnostic assays. Hybrid platforms where discrete components are combined in a single platform are a suitable approach to integration, where combining multiple device fabrication steps on a single substrate is not possible due to incompatible or costly fabrication steps. We integrate three devices each with a specific system functionality: (i) a silicon electro-wetting-on-dielectric (EWOD) device to move and mix sample and reagent droplets in an oil phase, (ii) a polymer microfluidic chip containing channels and reservoirs and (iii) an aqueous phase glass microarray for fluorescence microarray hybridization detection. The EWOD device offers the possibility of fully integrating on-chip sample preparation using nanolitre sample and reagent volumes. A key challenge is sample transfer from the oil phase EWOD device to the aqueous phase microarray for hybridization detection. The EWOD device, waveguide performance and functionality are maintained during the integration process. An on-chip biochemical protocol for arrayed primer extension (APEX) was implemented for single nucleotide polymorphism (SNiP) analysis. The prepared sample is aspirated from the EWOD oil phase to the aqueous phase microarray for hybridization. A bench-top instrumentation system was also developed around the integrated platform to drive the EWOD electrodes, implement APEX sample heating and image the microarray after hybridization.

  16. Development of a digital microfluidic platform for point of care testing

    PubMed Central

    Sista, Ramakrishna; Hua, Zhishan; Thwar, Prasanna; Sudarsan, Arjun; Srinivasan, Vijay; Eckhardt, Allen; Pollack, Michael; Pamula, Vamsee

    2009-01-01

    Point of care testing is playing an increasingly important role in improving the clinical outcome in health care management. The salient features of a point of care device are quick results, integrated sample preparation and processing, small sample volumes, portability, multifunctionality and low cost. In this paper, we demonstrate some of these salient features utilizing an electrowetting-based Digital Microfluidic platform. We demonstrate the performance of magnetic bead-based immunoassays (cardiac troponin I) on a digital microfluidic cartridge in less than 8 minutes using whole blood samples. Using the same microfluidic cartridge, a 40-cycle real-time polymerase chain reaction was performed within 12 minutes by shuttling a droplet between two thermal zones. We further demonstrate, on the same cartridge, the capability to perform sample preparation for bacterial and fungal infectious disease pathogens (methicillin-resistance Staphylococcus aureus and Candida albicans) and for human genomic DNA using magnetic beads. In addition to rapid results and integrated sample preparation, electrowetting-based digital microfluidic instruments are highly portable because fluid pumping is performed electronically. All the digital microfluidic chips presented here were fabricated on printed circuit boards utilizing mass production techniques that keep the cost of the chip low. Due to the modularity and scalability afforded by digital microfluidics, multifunctional testing capability, such as combinations within and between immunoassays, DNA amplification, and enzymatic assays, can be brought to the point of care at a relatively low cost because a single chip can be configured in software for different assays required along the path of care. PMID:19023472

  17. Preparation of high-quality planar FeRh thin films for in situ TEM investigations

    NASA Astrophysics Data System (ADS)

    Almeida, Trevor P.; McGrouther, Damien; Pivak, Yevheniy; Perez Garza, Hector Hugo; Temple, Rowan; Massey, Jamie; Marrows, Christopher H.; McVitie, Stephen

    2017-10-01

    The preparation of a planar FeRh thin film using a focused ion beam (FIB) secondary electron microscope (SEM) for the purpose of in situ transmission electron microscopy (TEM) is presented. A custom SEM stub with 45° faces allows for the transfer and milling of the sample on a TEM heating chip, whilst Fresnel imaging within the TEM revealed the presence of the magnetic domain walls, confirming the quality of the FIB-prepared sample.

  18. Microfluidic integration of parallel solid-phase liquid chromatography.

    PubMed

    Huft, Jens; Haynes, Charles A; Hansen, Carl L

    2013-03-05

    We report the development of a fully integrated microfluidic chromatography system based on a recently developed column geometry that allows for robust packing of high-performance separation columns in poly(dimethylsiloxane) microfluidic devices having integrated valves made by multilayer soft lithography (MSL). The combination of parallel high-performance separation columns and on-chip plumbing was used to achieve a fully integrated system for on-chip chromatography, including all steps of automated sample loading, programmable gradient generation, separation, fluorescent detection, and sample recovery. We demonstrate this system in the separation of fluorescently labeled DNA and parallel purification of reverse transcription polymerase chain reaction (RT-PCR) amplified variable regions of mouse immunoglobulin genes using a strong anion exchange (AEX) resin. Parallel sample recovery in an immiscible oil stream offers the advantage of low sample dilution and high recovery rates. The ability to perform nucleic acid size selection and recovery on subnanogram samples of DNA holds promise for on-chip genomics applications including sequencing library preparation, cloning, and sample fractionation for diagnostics.

  19. Capacitive deionization on-chip as a method for microfluidic sample preparation.

    PubMed

    Roelofs, Susan H; Kim, Bumjoo; Eijkel, Jan C T; Han, Jongyoon; van den Berg, Albert; Odijk, Mathieu

    2015-03-21

    Desalination as a sample preparation step is essential for noise reduction and reproducibility of mass spectrometry measurements. A specific example is the analysis of proteins for medical research and clinical applications. Salts and buffers that are present in samples need to be removed before analysis to improve the signal-to-noise ratio. Capacitive deionization is an electrostatic desalination (CDI) technique which uses two porous electrodes facing each other to remove ions from a solution. Upon the application of a potential of 0.5 V ions migrate to the electrodes and are stored in the electrical double layer. In this article we demonstrate CDI on a chip, and desalinate a solution by the removal of 23% of Na(+) and Cl(-) ions, while the concentration of a larger molecule (FITC-dextran) remains unchanged. For the first time impedance spectroscopy is introduced to monitor the salt concentration in situ in real-time in between the two desalination electrodes.

  20. Clinically relevant advances in on-chip affinity-based electrophoresis and electrochromatography.

    PubMed

    Hou, Chenlu; Herr, Amy E

    2008-08-01

    Clinical and point-of-care disease diagnostics promise to play an important role in personalized medicine, new approaches to global health, and health monitoring. Emerging instrument platforms based on lab-on-a-chip technology can confer performance advantages successfully exploited in electrophoresis and electrochromatography to affinity-based electrokinetic separations. This review surveys lab-on-a-chip diagnostic developments in affinity-based electrokinetic separations for quantitation of proteins, integration of preparatory functions needed for subsequent analysis of diverse biological samples, and initial forays into multiplexed analyses. The technologies detailed here underpin new clinical and point-of-care diagnostic strategies. The techniques and devices promise to advance translation of until now laboratory-based sample preparation and analytical assays to near-patient settings.

  1. Optimization of multiplexed PCR on an integrated microfluidic forensic platform for rapid DNA analysis.

    PubMed

    Estes, Matthew D; Yang, Jianing; Duane, Brett; Smith, Stan; Brooks, Carla; Nordquist, Alan; Zenhausern, Frederic

    2012-12-07

    This study reports the design, prototyping, and assay development of multiplexed polymerase chain reaction (PCR) on a plastic microfluidic device. Amplification of 17 DNA loci is carried out directly on-chip as part of a system for continuous workflow processing from sample preparation (SP) to capillary electrophoresis (CE). For enhanced performance of on-chip PCR amplification, improved control systems have been developed making use of customized Peltier assemblies, valve actuators, software, and amplification chemistry protocols. Multiple enhancements to the microfluidic chip design have been enacted to improve the reliability of sample delivery through the various on-chip modules. This work has been enabled by the encapsulation of PCR reagents into a solid phase material through an optimized Solid Phase Encapsulating Assay Mix (SPEAM) bead-based hydrogel fabrication process. SPEAM bead technology is reliably coupled with precise microfluidic metering and dispensing for efficient amplification and subsequent DNA short tandem repeat (STR) fragment analysis. This provides a means of on-chip reagent storage suitable for microfluidic automation, with the long shelf-life necessary for point-of-care (POC) or field deployable applications. This paper reports the first high quality 17-plex forensic STR amplification from a reference sample in a microfluidic chip with preloaded solid phase reagents, that is designed for integration with up and downstream processing.

  2. Optical biosensor system with integrated microfluidic sample preparation and TIRF based detection

    NASA Astrophysics Data System (ADS)

    Gilli, Eduard; Scheicher, Sylvia R.; Suppan, Michael; Pichler, Heinz; Rumpler, Markus; Satzinger, Valentin; Palfinger, Christian; Reil, Frank; Hajnsek, Martin; Köstler, Stefan

    2013-05-01

    There is a steadily growing demand for miniaturized bioanalytical devices allowing for on-site or point-of-care detection of biomolecules or pathogens in applications like diagnostics, food testing, or environmental monitoring. These, so called labs-on-a-chip or micro-total analysis systems (μ-TAS) should ideally enable convenient sample-in - result-out type operation. Therefore, the entire process from sample preparation, metering, reagent incubation, etc. to detection should be performed on a single disposable device (on-chip). In the early days such devices were mainly fabricated using glass or silicon substrates and adapting established fabrication technologies from the electronics and semiconductor industry. More recently, the development focuses on the use of thermoplastic polymers as they allow for low-cost high volume fabrication of disposables. One of the most promising materials for the development of plastic based lab-on-achip systems are cyclic olefin polymers and copolymers (COP/COC) due to their excellent optical properties (high transparency and low autofluorescence) and ease of processing. We present a bioanalytical system for whole blood samples comprising a disposable plastic chip based on TIRF (total internal reflection fluorescence) optical detection. The chips were fabricated by compression moulding of COP and microfluidic channels were structured by hot embossing. These microfluidic structures integrate several sample pretreatment steps. These are the separation of erythrocytes, metering of sample volume using passive valves, and reagent incubation for competitive bioassays. The surface of the following optical detection zone is functionalized with specific capture probes in an array format. The plastic chips comprise dedicated structures for simple and effective coupling of excitation light from low-cost laser diodes. This enables TIRF excitation of fluorescently labeled probes selectively bound to detection spots at the microchannel surface. The fluorescence of these detection arrays is imaged using a simple set-up based on a digital consumer camera. Image processing for spot detection and intensity calculation is accomplished using customized software. Using this combined TIRF excitation and imaging based detection approach allowes for effective suppression of background fluorescence from the sample, multiplexed detection in an array format, as well as internal calibration and background correction.

  3. Lab-on-chip systems for integrated bioanalyses

    PubMed Central

    Madaboosi, Narayanan; Soares, Ruben R.G.; Fernandes, João Tiago S.; Novo, Pedro; Moulas, Geraud; Chu, Virginia

    2016-01-01

    Biomolecular detection systems based on microfluidics are often called lab-on-chip systems. To fully benefit from the miniaturization resulting from microfluidics, one aims to develop ‘from sample-to-answer’ analytical systems, in which the input is a raw or minimally processed biological, food/feed or environmental sample and the output is a quantitative or qualitative assessment of one or more analytes of interest. In general, such systems will require the integration of several steps or operations to perform their function. This review will discuss these stages of operation, including fluidic handling, which assures that the desired fluid arrives at a specific location at the right time and under the appropriate flow conditions; molecular recognition, which allows the capture of specific analytes at precise locations on the chip; transduction of the molecular recognition event into a measurable signal; sample preparation upstream from analyte capture; and signal amplification procedures to increase sensitivity. Seamless integration of the different stages is required to achieve a point-of-care/point-of-use lab-on-chip device that allows analyte detection at the relevant sensitivity ranges, with a competitive analysis time and cost. PMID:27365042

  4. Blister pouches for effective reagent storage and release for low cost point-of-care diagnostic applications

    NASA Astrophysics Data System (ADS)

    Smith, Suzanne; Sewart, Rene; Land, Kevin; Roux, Pieter; Gärtner, Claudia; Becker, Holger

    2016-03-01

    Lab-on-a-chip devices are often applied to point-of-care diagnostic solutions as they are low-cost, compact, disposable, and require only small sample volumes. For such devices, various reagents are required for sample preparation and analysis and, for an integrated solution to be realized, on-chip reagent storage and automated introduction are required. This work describes the implementation and characterization of effective liquid reagent storage and release mechanisms utilizing blister pouches applied to various point-of-care diagnostic device applications. The manufacturing aspects as well as performance parameters are evaluated.

  5. Chromatin Immunoprecipitation (ChIP) Protocol for Low-abundance Embryonic Samples.

    PubMed

    Rehimi, Rizwan; Bartusel, Michaela; Solinas, Francesca; Altmüller, Janine; Rada-Iglesias, Alvaro

    2017-08-29

    Chromatin immunoprecipitation (ChIP) is a widely-used technique for mapping the localization of post-translationally modified histones, histone variants, transcription factors, or chromatin-modifying enzymes at a given locus or on a genome-wide scale. The combination of ChIP assays with next-generation sequencing (i.e., ChIP-Seq) is a powerful approach to globally uncover gene regulatory networks and to improve the functional annotation of genomes, especially of non-coding regulatory sequences. ChIP protocols normally require large amounts of cellular material, thus precluding the applicability of this method to investigating rare cell types or small tissue biopsies. In order to make the ChIP assay compatible with the amount of biological material that can typically be obtained in vivo during early vertebrate embryogenesis, we describe here a simplified ChIP protocol in which the number of steps required to complete the assay were reduced to minimize sample loss. This ChIP protocol has been successfully used to investigate different histone modifications in various embryonic chicken and adult mouse tissues using low to medium cell numbers (5 x 10 4 - 5 x 10 5 cells). Importantly, this protocol is compatible with ChIP-seq technology using standard library preparation methods, thus providing global epigenomic maps in highly relevant embryonic tissues.

  6. A Chip-Capillary Hybrid Device for Automated Transfer of Sample Pre-Separated by Capillary Isoelectric Focusing to Parallel Capillary Gel Electrophoresis for Two-Dimensional Protein Separation

    PubMed Central

    Lu, Joann J.; Wang, Shili; Li, Guanbin; Wang, Wei; Pu, Qiaosheng; Liu, Shaorong

    2012-01-01

    In this report, we introduce a chip-capillary hybrid device to integrate capillary isoelectric focusing (CIEF) with parallel capillary sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE) or capillary gel electrophoresis (CGE) toward automating two-dimensional (2D) protein separations. The hybrid device consists of three chips that are butted together. The middle chip can be moved between two positions to re-route the fluidic paths, which enables the performance of CIEF and injection of proteins partially resolved by CIEF to CGE capillaries for parallel CGE separations in a continuous and automated fashion. Capillaries are attached to the other two chips to facilitate CIEF and CGE separations and to extend the effective lengths of CGE columns. Specifically, we illustrate the working principle of the hybrid device, develop protocols for producing and preparing the hybrid device, and demonstrate the feasibility of using this hybrid device for automated injection of CIEF-separated sample to parallel CGE for 2D protein separations. Potentials and problems associated with the hybrid device are also discussed. PMID:22830584

  7. A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins

    NASA Astrophysics Data System (ADS)

    Ahmad, Habib; Sutherland, Alex; Shin, Young Shik; Hwang, Kiwook; Qin, Lidong; Krom, Russell-John; Heath, James R.

    2011-09-01

    Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells.

  8. A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins

    PubMed Central

    Ahmad, Habib; Sutherland, Alex; Shin, Young Shik; Hwang, Kiwook; Qin, Lidong; Krom, Russell-John; Heath, James R.

    2011-01-01

    Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells. PMID:21974603

  9. A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins.

    PubMed

    Ahmad, Habib; Sutherland, Alex; Shin, Young Shik; Hwang, Kiwook; Qin, Lidong; Krom, Russell-John; Heath, James R

    2011-09-01

    Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells. © 2011 American Institute of Physics

  10. Evaluation of the thermal conductance of flip-chip bonding structure utilizing the measurement based on Fourier's law of heat conduction at steady-state

    NASA Astrophysics Data System (ADS)

    Wu, Chia-Yu; Huang, Yin-Hsien; Wu, Hsin-Han; Hsieh, Tsung-Eong

    2018-06-01

    Fourier's law of heat conduction at steady-state was adopted to establish a measurement method utilizing platinum (Pt) thin-film electrodes as the heater and the temperature sensor. The thermal conductivities (κ's) of Pyrex glass, an epoxy resin and a commercial underfill for flip-chip devices were measured and a good agreement with previously reported values was obtained. The thermal boundary resistances (RTBR's) of Pt/sample interfaces were also extracted for discussing their influence on the thermal conduction of samples. Afterward, the flip-chip samples with 2×2 solder joint array utilizing Si wafers as the die and the substrate, without and with the underfills, were prepared and their thermal conductance were measured. For the sample without underfill, the air presenting in the gap of die and the substrate led to the poor thermal conductance of sample. With the insertion of underfills, the thermal conductance of flip-chip samples improved. The resistance to heat transfer across Si/underfill interfaces was also suppressed and to promote the thermal conductance of samples. The thermal properties of underfill and RTBR at Si/underfill interface were further implanted in the calculation of thermal conductance of flip-chip samples containing various solder joint arrays. The increasing number of solder joints diminished the influence of thermal conduction of underfill and RTBR of Si/underfill interface on the thermal conductance of samples. The insertion of underfill with high-κ value might promote the heat conductance of samples containing low-density solder joint arrays; however, it became insignificant in improving the heat conductance of samples containing high-density solder joint arrays.

  11. Preparation of a Superhydrophobic and Peroxidase-like Activity Array Chip for H2O2 Sensing by Surface-Enhanced Raman Scattering.

    PubMed

    Yu, Zhi; Park, Yeonju; Chen, Lei; Zhao, Bing; Jung, Young Mee; Cong, Qian

    2015-10-28

    In this paper, we propose a novel and simple method for preparing a dual-biomimetic functional array possessing both superhydrophobic and peroxidase-like activity that can be used for hydrogen peroxide (H2O2) sensing. The proposed method is an integration innovation that combines the above two properties and surface-enhanced Raman scattering (SERS). We integrated a series of well-ordered arrays of Au points (d = 1 mm) onto a superhydrophobic copper (Cu)/silver (Ag) surface by replicating an arrayed molybdenum template. Instead of using photoresists and the traditional lithography method, we utilized a chemical etching method (a substitution reaction between Cu and HAuCl4) with a Cu/Ag superhydrophobic surface as the barrier layer, which has the benefit of water repellency. The as-prepared Au points were observed to possess peroxidase-like activity, allowing for catalytic oxidation of the chromogenic molecule o-phenylenediamine dihydrochloride (OPD). Oxidation was evidenced by a color change in the presence of H2O2, which allows the array chip to act as an H2O2 sensor. In this study, the water repellency of the superhydrophobic surface was used to fabricate the array chip and increase the local reactant concentration during the catalytic reaction. As a result, the catalytic reaction occurred when only 2 μL of an aqueous sample (OPD/H2O2) was placed onto the Au point, and the enzymatic product, 2,3-diaminophenazine, showed a SERS signal distinguishable from that of OPD after mixing with 2 μL of colloidal Au. Using the dual-biomimetic functional array chip, quantitative analysis of H2O2 was performed by observing the change in the SERS spectra, which showed a concentration-dependent behavior for H2O2. This method allows for the detection of H2O2 at concentrations as low as 3 pmol per 2 μL of sample, which is a considerable advantage in H2O2 analysis. The as-prepared substrate was convenient for H2O2 detection because only a small amount of sample was required in each analysis. Highly sensitive detection was realized using SERS. Therefore, this chip was shown to exhibit significant potential for applications in bioanalysis.

  12. Lab-on-a-chip technologies for genodermatoses: Recent progress and future perspectives.

    PubMed

    Hongzhou, Cui; Shuping, Guo; Wenju, Wang; Li, Li; Lulu, Wei; Linjun, Deng; Jingmin, Li; Xiaoli, Ren; Li, Bai

    2017-02-01

    In recent years, molecular biology has proven to be a great asset in our understanding of mechanisms in genodermatoses. However, bench to bedside translation research lags far behind. Advances in lab-on-a-chip technologies enabled programmable, reconfigurable, and scalable manipulation of a variety of laboratory procedures. Sample preparation, microfluidic reactions, and continuous monitoring systems can be integrated on a small chip. These advantages have attracted attention in various fields of clinical application including diagnosis of inherited skin diseases. This review lists an overview of the underlying genes and mutations and describes prospective application of lab-on-a-chip technologies as solutions to challenges for point-of-care genodematoses diagnosis. Copyright © 2016. Published by Elsevier B.V.

  13. Integration of reconfigurable potentiometric electrochemical sensors into a digital microfluidic platform.

    PubMed

    Farzbod, Ali; Moon, Hyejin

    2018-05-30

    This paper presents the demonstration of on-chip fabrication of a potassium-selective sensor array enabled by electrowetting on dielectric digital microfluidics for the first time. This demonstration proves the concept that electrochemical sensors can be seamlessly integrated with sample preparation units in a digital microfluidic platform. More significantly, the successful on-chip fabrication of a sensor array indicates that sensors become reconfigurable and have longer lifetime in a digital microfluidic platform. The on-chip fabrication of ion-selective electrodes includes electroplating Ag followed by forming AgCl layer by chemical oxidation and depositing a thin layer of desired polymer-based ion selective membrane on one of the sensor electrodes. In this study, potassium ionophores work as potassium ion channels and make the membrane selective to potassium ions. This selectiveness results in the voltage difference across the membrane layer, which is correlated with potassium ion concentration. The calibration curve of the fabricated potassium-selective electrode demonstrates the slope of 58 mV/dec for potassium concentration in KCl sample solutions and shows good agreement with the ideal Nernstian response. The proposed sensor platform is an outstanding candidate for a portable home-use for continuous monitoring of ions thanks to its advantages such as easy automation of sample preparation and detection processes, elongated sensor lifetime, minimal membrane and sample consumption, and user-definable/reconfigurable sensor array. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. A pilot study of gene expression analysis in workers with hand-arm vibration syndrome.

    PubMed

    Maeda, Setsuo; Yu, Xiaozhong; Wang, Rui-Sheng; Sakakibara, Hisataka

    2008-04-01

    The purpose of this pilot study was to examine differences in gene expressions by cDNA microarray analysis of hand-arm vibration syndrome (HAVS) patients. Vein blood samples were collected and total RNA was extracted. All blood samples were obtained in the morning in one visit after a standard light breakfast. We performed microarray analysis with the labeled cDNA prepared by reverse transcription from RNA samples, using the Human CHIP version 1 (DNA Chip Research Inc, Yokohama, Japan). There are 2,976 genes on the chip, and these genes were selected from a cDNA library prepared with human peripheral white blood cells (WBC). Different gene levels between the HAVS patients and controls, and between groups of HAVS with different levels of symptoms, were indicated by the randomized variance model. The most up-regulated genes were analyzed for their possible functions and association with the occurrence of HAVS. From the results of this pilot study, although the results were obtained a limited number of subjects, it would appear that cDNA microarray analysis of HAVS patients has potential as a new objective method of HAVS diagnosis. Further research is needed to examine the gene expression with increased numbers of patients at different stages of HAVS.

  15. Multifunctional Au NPs-polydopamine-polyvinylidene fluoride membrane chips as probe for enrichment and rapid detection of organic contaminants.

    PubMed

    Wang, Saihua; Niu, Hongyun; Cai, Yaqi; Cao, Dong

    2018-05-01

    High-throughput and rapid detection of hazardous compounds in complicated samples is essential for the solution of environmental problems. We have prepared a "pH-paper-like" chip which can rapidly "indicate" the occurrence of organic contaminants just through dipping the chip in water samples for short time followed by fast analysis with surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF MS). The chips are composed of polyvinylidene fluoride membrane (PVDFM), polydopamine (PDA) film and Au nanoparticles (Au NPs), which are layer-by-layer assembled according to the adhesion, self-polymerization and reduction property of dopamine. In the Au NPs loaded polydopamine-polyvinylidene fluoride membrane (Au NPs-PDA-PVDFM) chips, PVDFM combined with PDA film are responsible for the enrichment of organic analyte through hydrophobic interactions and π-π stacking; Au NPs serve as effective SALDI matrix for the rapid detection of target analyte. After dipping into water solution for minutes, the Au-PDA-PVDFM chips with enriched organic analytes can be detected directly with SALDI-TOF MS. The good solid-phase extraction performance of the PDA-PVDFM components, remarkable matrix effect of the loaded AuNPs, and sensitivity of the SALDI-TOF MS technique ensure excellent sensitivity and reproducibility for the quantification of trace levels of organic contaminants in environmental water samples. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. The use of fruit extracts for production of apple chips with enhanced antioxidant activity

    PubMed

    Tarko, Tomasz; Duda-Chodak, Aleksandra; Semik-Szczurak, Dorota

    Style and pace of life make consumers more willing to reach for snack products. This group of processed food includes, among others, fruit chips. Due to the increasing incidence of diseases associated with the excessive exposure to free radicals foods enriched with antioxidant compounds, eg. polyphenols, can be introduced into the sale. The aim of the study was to use the fruit extracts for the production of apple chips with enhanced antioxidant activity. ‘Golden Delicious’ variety of apple fruit was used to produce chips. Apple chips were prepared by slicing, soaking in a sugar solution and pre-drying in a microwave oven. Chips were enriched with extracts prepared from fruits of chokeberry, five-flavor berry, Cornelian cherry, woodland hawthorn, goji berry, Japanese quince and cranberry microcarpa. For this purpose, pre-dried apple slices were soaked (5 min) in ethanolic extract of fruits and then dried to achieve a 5% moisture content. Chips were sensory evaluated and their antioxidant activity and total polyphenols content were determined. All enriched apple chips were characterized by high antioxidant activity and a relatively high value of total polyphenols content. Chips soaked in extracts of five-flavor berry, cranberry and goji berry were characterized by the highest antioxidant potential. Samples obtained by using chokeberry and Cornelian cherry extracts showed the highest content of polyphenols. High sensory attractiveness of enriched chips was also showed. The chips with the addition of fiveflavor berry extract were exceptions. Their taste was not acceptable. Fruit extracts are a valuable material for chips enrichment. Taking into account all the analyzed differentiators, extracts of Japanese quince, goji berry and woodland hawthorn were found to be the best enriching additives. The chips soaked in extract of five-flavor berry, despite their high antioxidant activity, were disqualified due to very low score of sensory evaluation.

  17. MOLECULAR METHODS (E.G., MICROARRAYS) APPLIED TO PLANT GENOMES FOR ASSESSING GENETIC CHANGE AND ENVIRONMENTAL STRESS

    EPA Science Inventory

    This is a technical document that presents a detailed sample standard operating procedure (S.O.P.) for preparing plant nucleic acid samples for microarray analyses using commercial ¿chips¿ such as those sold by Affymetrix. It also presents the application of a commercially availa...

  18. High-throughput automated microfluidic sample preparation for accurate microbial genomics

    PubMed Central

    Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B.; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P.; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C.

    2017-01-01

    Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications. PMID:28128213

  19. Microfluidic cell disruption system employing a magnetically actuated diaphragm.

    PubMed

    Huh, Yun Suk; Choi, Jong Hyun; Huh, Kyoung Ae Kim; Kim, Kyoung Ae; Park, Tae Jung; Hong, Yeon Ki; Kim, Do Hyun; Hong, Won Hi; Lee, Sang Yup

    2007-12-01

    A microfluidic cell lysis chip equipped with a micromixer and SPE unit was developed and used for quantitative analysis of intracellular proteins. This miniaturized sample preparation system can be employed for any purpose where cell disruption is needed to obtain intracellular constituents for the subsequent analysis. This system comprises a magnetically actuated micromixer to disrupt cells, a hydrophobic valve to manipulate the cell lysate, and a packed porous polymerized monolith chamber for SPE and filtering debris from the cell lysate. Using recombinant Escherichia coli expressing intracellular enhanced green fluorescent protein (EGFP) and lipase as model bacteria, we optimized the cell disruption condition with respect to the lysis buffer composition, mixing time, and the frequency of the diaphragm in the micromixer, which was magnetically actuated by an external magnetic stirrer in the micromixer chamber. The lysed sample prepared under the optimal condition was purified by the packed SPE in the microfluidic chip. At a frequency of 1.96 Hz, the final cell lysis efficiency and relative fluorescence intensity of EGFP after the cell disruption process were greater than 90 and 94%, respectively. Thus, this microfluidic cell disruption chip can be used for the efficient lysis of cells for further analysis of intracellular contents in many applications.

  20. Laser Light-field Fusion for Wide-field Lensfree On-chip Phase Contrast Microscopy of Nanoparticles

    NASA Astrophysics Data System (ADS)

    Kazemzadeh, Farnoud; Wong, Alexander

    2016-12-01

    Wide-field lensfree on-chip microscopy, which leverages holography principles to capture interferometric light-field encodings without lenses, is an emerging imaging modality with widespread interest given the large field-of-view compared to lens-based techniques. In this study, we introduce the idea of laser light-field fusion for lensfree on-chip phase contrast microscopy for detecting nanoparticles, where interferometric laser light-field encodings acquired using a lensfree, on-chip setup with laser pulsations at different wavelengths are fused to produce marker-free phase contrast images of particles at the nanometer scale. As a proof of concept, we demonstrate, for the first time, a wide-field lensfree on-chip instrument successfully detecting 300 nm particles across a large field-of-view of ~30 mm2 without any specialized or intricate sample preparation, or the use of synthetic aperture- or shift-based techniques.

  1. Laser Light-field Fusion for Wide-field Lensfree On-chip Phase Contrast Microscopy of Nanoparticles.

    PubMed

    Kazemzadeh, Farnoud; Wong, Alexander

    2016-12-13

    Wide-field lensfree on-chip microscopy, which leverages holography principles to capture interferometric light-field encodings without lenses, is an emerging imaging modality with widespread interest given the large field-of-view compared to lens-based techniques. In this study, we introduce the idea of laser light-field fusion for lensfree on-chip phase contrast microscopy for detecting nanoparticles, where interferometric laser light-field encodings acquired using a lensfree, on-chip setup with laser pulsations at different wavelengths are fused to produce marker-free phase contrast images of particles at the nanometer scale. As a proof of concept, we demonstrate, for the first time, a wide-field lensfree on-chip instrument successfully detecting 300 nm particles across a large field-of-view of ~30 mm 2 without any specialized or intricate sample preparation, or the use of synthetic aperture- or shift-based techniques.

  2. A comparative study of ChIP-seq sequencing library preparation methods.

    PubMed

    Sundaram, Arvind Y M; Hughes, Timothy; Biondi, Shea; Bolduc, Nathalie; Bowman, Sarah K; Camilli, Andrew; Chew, Yap C; Couture, Catherine; Farmer, Andrew; Jerome, John P; Lazinski, David W; McUsic, Andrew; Peng, Xu; Shazand, Kamran; Xu, Feng; Lyle, Robert; Gilfillan, Gregor D

    2016-10-21

    ChIP-seq is the primary technique used to investigate genome-wide protein-DNA interactions. As part of this procedure, immunoprecipitated DNA must undergo "library preparation" to enable subsequent high-throughput sequencing. To facilitate the analysis of biopsy samples and rare cell populations, there has been a recent proliferation of methods allowing sequencing library preparation from low-input DNA amounts. However, little information exists on the relative merits, performance, comparability and biases inherent to these procedures. Notably, recently developed single-cell ChIP procedures employing microfluidics must also employ library preparation reagents to allow downstream sequencing. In this study, seven methods designed for low-input DNA/ChIP-seq sample preparation (Accel-NGS® 2S, Bowman-method, HTML-PCR, SeqPlex™, DNA SMART™, TELP and ThruPLEX®) were performed on five replicates of 1 ng and 0.1 ng input H3K4me3 ChIP material, and compared to a "gold standard" reference PCR-free dataset. The performance of each method was examined for the prevalence of unmappable reads, amplification-derived duplicate reads, reproducibility, and for the sensitivity and specificity of peak calling. We identified consistent high performance in a subset of the tested reagents, which should aid researchers in choosing the most appropriate reagents for their studies. Furthermore, we expect this work to drive future advances by identifying and encouraging use of the most promising methods and reagents. The results may also aid judgements on how comparable are existing datasets that have been prepared with different sample library preparation reagents.

  3. Development of a magnetic lab-on-a-chip for point-of-care sepsis diagnosis

    NASA Astrophysics Data System (ADS)

    Schotter, Joerg; Shoshi, Astrit; Brueckl, Hubert

    2009-05-01

    We present design criteria, operation principles and experimental examples of magnetic marker manipulation for our magnetic lab-on-a-chip prototype. It incorporates both magnetic sample preparation and detection by embedded GMR-type magnetoresistive sensors and is optimized for the automated point-of-care detection of four different sepsis-indicative cytokines directly from about 5 μl of whole blood. The sample volume, magnetic particle size and cytokine concentration determine the microfluidic volume, sensor size and dimensioning of the magnetic gradient field generators. By optimizing these parameters to the specific diagnostic task, best performance is expected with respect to sensitivity, analysis time and reproducibility.

  4. Development of the Large-Scale Oligonucleotide Chip for the Diagnosis of Plant Viruses and its Practical Use

    PubMed Central

    Nam, Moon; Kim, Jeong-Seon; Lim, Seungmo; Park, Chung Youl; Kim, Jeong-Gyu; Choi, Hong-Soo; Lim, Hyoun-Sub; Moon, Jae Sun; Lee, Su-Heon

    2014-01-01

    A large-scale oligonucleotide (LSON) chip was developed for the detection of the plant viruses with known genetic information. The LSON chip contains two sets of 3,978 probes for 538 species of targets including plant viruses, satellite RNAs and viroids. A hundred forty thousand probes, consisting of isolate-, species- and genus-specific probes respectively, are designed from 20,000 of independent nucleotide sequence of plant viruses. Based on the economic importance, the amount of genome information, and the number of strains and/or isolates, one to fifty-one probes for each target virus are selected and spotted on the chip. The standard and field samples for the analysis of the LSON chip have been prepared and tested by RT-PCR. The probe’s specific and/or nonspecific reaction patterns by LSON chip allow us to diagnose the unidentified viruses. Thus, the LSON chip in this study could be highly useful for the detection of unexpected plant viruses, the monitoring of emerging viruses and the fluctuation of the population of major viruses in each plant. PMID:25288985

  5. Disposable MoS2-Arrayed MALDI MS Chip for High-Throughput and Rapid Quantification of Sulfonamides in Multiple Real Samples.

    PubMed

    Zhao, Yaju; Tang, Minmin; Liao, Qiaobo; Li, Zhoumin; Li, Hui; Xi, Kai; Tan, Li; Zhang, Mei; Xu, Danke; Chen, Hong-Yuan

    2018-04-27

    In this work, we demonstrate, for the first time, the development of a disposable MoS 2 -arrayed matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) chip combined with an immunoaffinity enrichment method for high-throughput, rapid, and simultaneous quantitation of multiple sulfonamides (SAs). The disposable MALDI MS chip was designed and fabricated by MoS 2 array formation on a commercial indium tin oxide (ITO) glass slide. A series of SAs were analyzed, and clear deprotonated signals were obtained in negative-ion mode. Compared with MoS 2 -arrayed commercial steel plate, the prepared MALDI MS chip exhibited comparable LDI efficiency, providing a good alternative and disposable substrate for MALDI MS analysis. Furthermore, internal standard (IS) was previously deposited onto the MoS 2 array to simplify the experimental process for MALDI MS quantitation. 96 sample spots could be analyzed within 10 min in one single chip to perform quantitative analysis, recovery studies, and real foodstuff detection. Upon targeted extraction and enrichment by antibody conjugated magnetic beads, five SAs were quantitatively determined by the IS-first method with the linear range of 0.5-10 ng/mL ( R 2 > 0.990). Good recoveries and repeatability were obtained for spiked pork, egg, and milk samples. SAs in several real foodstuffs were successfully identified and quantified. The developed method may provide a promising tool for the routine analysis of antibiotic residues in real samples.

  6. Sequencing of real-world samples using a microfabricated hybrid device having unconstrained straight separation channels.

    PubMed

    Liu, Shaorong; Elkin, Christopher; Kapur, Hitesh

    2003-11-01

    We describe a microfabricated hybrid device that consists of a microfabricated chip containing multiple twin-T injectors attached to an array of capillaries that serve as the separation channels. A new fabrication process was employed to create two differently sized round channels in a chip. Twin-T injectors were formed by the smaller round channels that match the bore of the separation capillaries and separation capillaries were incorporated to the injectors through the larger round channels that match the outer diameter of the capillaries. This allows for a minimum dead volume and provides a robust chip/capillary interface. This hybrid design takes full advantage, such as sample stacking and purification and uniform signal intensity profile, of the unique chip injection scheme for DNA sequencing while employing long straight capillaries for the separations. In essence, the separation channel length is optimized for both speed and resolution since it is unconstrained by chip size. To demonstrate the reliability and practicality of this hybrid device, we sequenced over 1000 real-world samples from Human Chromosome 5 and Ciona intestinalis, prepared at Joint Genome Institute. We achieved average Phred20 read of 675 bases in about 70 min with a success rate of 91%. For the similar type of samples on MegaBACE 1000, the average Phred20 read is about 550-600 bases in 120 min separation time with a success rate of about 80-90%.

  7. Chemical and biological threat-agent detection using electrophoresis-based lab-on-a-chip devices.

    PubMed

    Borowsky, Joseph; Collins, Greg E

    2007-10-01

    The ability to separate complex mixtures of analytes has made capillary electrophoresis (CE) a powerful analytical tool since its modern configuration was first introduced over 25 years ago. The technique found new utility with its application to the microfluidics based lab-on-a-chip platform (i.e., microchip), which resulted in ever smaller footprints, sample volumes, and analysis times. These features, coupled with the technique's potential for portability, have prompted recent interest in the development of novel analyzers for chemical and biological threat agents. This article will comment on three main areas of microchip CE as applied to the separation and detection of threat agents: detection techniques and their corresponding limits of detection, sampling protocol and preparation time, and system portability. These three areas typify the broad utility of lab-on-a-chip for meeting critical, present-day security, in addition to illustrating areas wherein advances are necessary.

  8. Lab-on-a-chip sensor for measuring Zn by stripping voltammetry

    NASA Astrophysics Data System (ADS)

    Pei, Xing; Kang, Wenjing; Yue, Wei; Bange, Adam; Wong, Hector R.; Heineman, William R.; Papautsky, Ian

    2012-03-01

    This work reports on continuing development of a lab-on-a-chip sensor for electrochemical detection of heavy metal zinc in blood serum. The sensor consists of a three electrode system, including an environmentally-friendly bismuth working electrode, a Ag/AgCl reference electrode, and a gold auxiliary electrode. By optimizing the electrodeposition of bismuth film, better control of fabrication steps and improving interface between the sensor and potentiostat, repeatability and sensitivity of the lab-on-a-chip sensor has been improved. Through optimization of electrolyte and stripping voltammetry parameters, limits of detection were greatly improved. The optimized sensor was able to measure zinc in in the physiological range of 65-95 μg/dL. Ultimately, with further development and integrated sample preparation sensor system will permit rapid (min) measurements of zinc from a sub-mL sample (a few drops of blood) for bedside monitoring.

  9. Integrated microfluidic systems for sample preparation and detection of respiratory pathogen Bordetella pertussis.

    PubMed

    de la Rosa, Carlos; Prakash, Ranjit; Tilley, Peter A; Fox, Julie D; Kaler, Karan V i S

    2007-01-01

    An integrated microfluidic system for combined manipulation, pre-concentration, and lysis of samples containing Bordetella pertussis by dielectrophoresis and electroporation has been developed and implemented. The microfluidic device was able to pre-concentrate the amount of B. pertussis cells present in 200 microl of a B. pertussis suspension stock into a 20 microl volume. The device exhibited optimal sample pre-concentration of 6.7x at a stock value of 10(3) cfu/ml and at a flow rate of 250 microl/h. Electro-disruption experiments showed that on-chip-based electroporation is an effective solution for lysis of B. pertussis cells that is easily integrated with dielectrophoresis assisted pre-concentration procedures. Pulsed voltage applied, number of pulses, and presence of potassium chloride in a B. pertussis suspension showed a reduction in B. pertussis cell viability by electroporation; and transmission electron microscopy confirmed B. pertussis cell disruption by electroporation. Genetic amplification and detection of the pre-concentrated sample employing an integrated chip-based system demonstrated a complete chip approach for pathogen detection.

  10. Progress in ion torrent semiconductor chip based sequencing.

    PubMed

    Merriman, Barry; Rothberg, Jonathan M

    2012-12-01

    In order for next-generation sequencing to become widely used as a diagnostic in the healthcare industry, sequencing instrumentation will need to be mass produced with a high degree of quality and economy. One way to achieve this is to recast DNA sequencing in a format that fully leverages the manufacturing base created for computer chips, complementary metal-oxide semiconductor chip fabrication, which is the current pinnacle of large scale, high quality, low-cost manufacturing of high technology. To achieve this, ideally the entire sensory apparatus of the sequencer would be embodied in a standard semiconductor chip, manufactured in the same fab facilities used for logic and memory chips. Recently, such a sequencing chip, and the associated sequencing platform, has been developed and commercialized by Ion Torrent, a division of Life Technologies, Inc. Here we provide an overview of this semiconductor chip based sequencing technology, and summarize the progress made since its commercial introduction. We described in detail the progress in chip scaling, sequencing throughput, read length, and accuracy. We also summarize the enhancements in the associated platform, including sample preparation, data processing, and engagement of the broader development community through open source and crowdsourcing initiatives. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. RNA–protein binding kinetics in an automated microfluidic reactor

    PubMed Central

    Ridgeway, William K.; Seitaridou, Effrosyni; Phillips, Rob; Williamson, James R.

    2009-01-01

    Microfluidic chips can automate biochemical assays on the nanoliter scale, which is of considerable utility for RNA–protein binding reactions that would otherwise require large quantities of proteins. Unfortunately, complex reactions involving multiple reactants cannot be prepared in current microfluidic mixer designs, nor is investigation of long-time scale reactions possible. Here, a microfluidic ‘Riboreactor’ has been designed and constructed to facilitate the study of kinetics of RNA–protein complex formation over long time scales. With computer automation, the reactor can prepare binding reactions from any combination of eight reagents, and is optimized to monitor long reaction times. By integrating a two-photon microscope into the microfluidic platform, 5-nl reactions can be observed for longer than 1000 s with single-molecule sensitivity and negligible photobleaching. Using the Riboreactor, RNA–protein binding reactions with a fragment of the bacterial 30S ribosome were prepared in a fully automated fashion and binding rates were consistent with rates obtained from conventional assays. The microfluidic chip successfully combines automation, low sample consumption, ultra-sensitive fluorescence detection and a high degree of reproducibility. The chip should be able to probe complex reaction networks describing the assembly of large multicomponent RNPs such as the ribosome. PMID:19759214

  12. A lab-on-a-chip system with integrated sample preparation and loop-mediated isothermal amplification for rapid and quantitative detection of Salmonella spp. in food samples.

    PubMed

    Sun, Yi; Quyen, Than Linh; Hung, Tran Quang; Chin, Wai Hoe; Wolff, Anders; Bang, Dang Duong

    2015-04-21

    Foodborne disease is a major public health threat worldwide. Salmonellosis, an infectious disease caused by Salmonella spp., is one of the most common foodborne diseases. Isolation and identification of Salmonella by conventional bacterial culture or molecular-based methods are time consuming and usually take a few hours to days to complete. In response to the demand for rapid on line or on site detection of pathogens, in this study, we describe for the first time an eight-chamber lab-on-a-chip (LOC) system with integrated magnetic bead-based sample preparation and loop-mediated isothermal amplification (LAMP) for rapid and quantitative detection of Salmonella spp. in food samples. The whole diagnostic procedures including DNA isolation, isothermal amplification, and real-time detection were accomplished in a single chamber. Up to eight samples could be handled simultaneously and the system was capable to detect Salmonella at concentration of 50 cells per test within 40 min. The simple design, together with high level of integration, isothermal amplification, and quantitative analysis of multiple samples in short time, will greatly enhance the practical applicability of the LOC system for rapid on-site screening of Salmonella for applications in food safety control, environmental surveillance, and clinical diagnostics.

  13. Automated imaging of cellular spheroids with selective plane illumination microscopy on a chip (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Paiè, Petra; Bassi, Andrea; Bragheri, Francesca; Osellame, Roberto

    2017-02-01

    Selective plane illumination microscopy (SPIM) is an optical sectioning technique that allows imaging of biological samples at high spatio-temporal resolution. Standard SPIM devices require dedicated set-ups, complex sample preparation and accurate system alignment, thus limiting the automation of the technique, its accessibility and throughput. We present a millimeter-scaled optofluidic device that incorporates selective plane illumination and fully automatic sample delivery and scanning. To this end an integrated cylindrical lens and a three-dimensional fluidic network were fabricated by femtosecond laser micromachining into a single glass chip. This device can upgrade any standard fluorescence microscope to a SPIM system. We used SPIM on a CHIP to automatically scan biological samples under a conventional microscope, without the need of any motorized stage: tissue spheroids expressing fluorescent proteins were flowed in the microchannel at constant speed and their sections were acquired while passing through the light sheet. We demonstrate high-throughput imaging of the entire sample volume (with a rate of 30 samples/min), segmentation and quantification in thick (100-300 μm diameter) cellular spheroids. This optofluidic device gives access to SPIM analyses to non-expert end-users, opening the way to automatic and fast screening of a high number of samples at subcellular resolution.

  14. Droplet-based biosensing for lab-on-a-chip, open microfluidics platforms

    USDA-ARS?s Scientific Manuscript database

    Low cost, portable sensors can transform health care by bringing easily available diagnostic devices to low and middle income population, particularly in developing countries. Sample preparation, analyte handling and labeling are primary cost concerns for traditional lab-based diagnostic systems. La...

  15. An integrated microfluidic analysis microsystems with bacterial capture enrichment and in-situ impedance detection

    NASA Astrophysics Data System (ADS)

    Liu, Hai-Tao; Wen, Zhi-Yu; Xu, Yi; Shang, Zheng-Guo; Peng, Jin-Lan; Tian, Peng

    2017-09-01

    In this paper, an integrated microfluidic analysis microsystems with bacterial capture enrichment and in-situ impedance detection was purposed based on microfluidic chips dielectrophoresis technique and electrochemical impedance detection principle. The microsystems include microfluidic chip, main control module, and drive and control module, and signal detection and processing modulet and result display unit. The main control module produce the work sequence of impedance detection system parts and achieve data communication functions, the drive and control circuit generate AC signal which amplitude and frequency adjustable, and it was applied on the foodborne pathogens impedance analysis microsystems to realize the capture enrichment and impedance detection. The signal detection and processing circuit translate the current signal into impendence of bacteria, and transfer to computer, the last detection result is displayed on the computer. The experiment sample was prepared by adding Escherichia coli standard sample into chicken sample solution, and the samples were tested on the dielectrophoresis chip capture enrichment and in-situ impedance detection microsystems with micro-array electrode microfluidic chips. The experiments show that the Escherichia coli detection limit of microsystems is 5 × 104 CFU/mL and the detection time is within 6 min in the optimization of voltage detection 10 V and detection frequency 500 KHz operating conditions. The integrated microfluidic analysis microsystems laid the solid foundation for rapid real-time in-situ detection of bacteria.

  16. A magnetic micropore chip for rapid (<1 hour) unbiased circulating tumor cell isolation and in situ RNA analysis.

    PubMed

    Ko, Jina; Bhagwat, Neha; Yee, Stephanie S; Black, Taylor; Redlinger, Colleen; Romeo, Janae; O'Hara, Mark; Raj, Arjun; Carpenter, Erica L; Stanger, Ben Z; Issadore, David

    2017-09-12

    The use of microtechnology for the highly selective isolation and sensitive detection of circulating tumor cells has shown enormous promise. One challenge for this technology is that the small feature sizes - which are the key to this technology's performance - can result in low sample throughput and susceptibility to clogging. Additionally, conventional molecular analysis of CTCs often requires cells to be taken off-chip for sample preparation and purification before analysis, leading to the loss of rare cells. To address these challenges, we have developed a microchip platform that combines fast, magnetic micropore based negative immunomagnetic selection (>10 mL h -1 ) with rapid on-chip in situ RNA profiling (>100× faster than conventional RNA labeling). This integrated chip can isolate both rare circulating cells and cell clusters directly from whole blood and allow individual cells to be profiled for multiple RNA cancer biomarkers, achieving sample-to-answer in less than 1 hour for 10 mL of whole blood. To demonstrate the power of this approach, we applied our device to the circulating tumor cell based diagnosis of pancreatic cancer. We used a genetically engineered lineage-labeled mouse model of pancreatic cancer (KPCY) to validate the performance of our chip. We show that in a cohort of patient samples (N = 25) that this device can detect and perform in situ RNA analysis on circulating tumor cells in patients with pancreatic cancer, even in those with extremely sparse CTCs (<1 CTC mL -1 of whole blood).

  17. Highly Sensitive, Label-Free Detection of 2,4-Dichlorophenoxyacetic Acid Using an Optofluidic Chip.

    PubMed

    Feng, Xueling; Zhang, Gong; Chin, Lip Ket; Liu, Ai Qun; Liedberg, Bo

    2017-07-28

    A highly sensitive approach for rapid and label-free detection of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) using an optofluidic chip is demonstrated. The optofluidic chip is prepared by covalent immobilization of 2,4-D-bovine serum albumin (2,4-D-BSA) conjugate to an integrated microring resonator. Subsequent detection of 2,4-D carried out in a competitive immunoreaction format enables selective detection of 2,4-D in different types of water samples, including bottled, tap, and lake water, at a limit of detection (LOD) of 4.5 pg/mL and in a quantitative range of 15-10 5 pg/mL. The microring resonator-based optofluidic chip is reusable with ultrahigh sensitivity that offers real-time and on-site detection of low-molecular-weight targets for potential applications in food safety and environmental monitoring.

  18. A Filtration-based Method of Preparing High-quality Nuclei from Cross-linked Skeletal Muscle for Chromatin Immunoprecipitation.

    PubMed

    Nohara, Kazunari; Chen, Zheng; Yoo, Seung-Hee

    2017-07-06

    Chromatin immunoprecipitation (ChIP) is a powerful method to determine protein binding to chromatin DNA. Fiber-rich skeletal muscle, however, has been a challenge for ChIP due to technical difficulty in isolation of high-quality nuclei with minimal contamination of myofibrils. Previous protocols have attempted to purify nuclei before cross-linking, which incurs the risk of altered DNA-protein interaction during the prolonged nuclei preparation process. In the current protocol, we first cross-linked the skeletal muscle tissue collected from mice, and the tissues were minced and sonicated. Since we found that ultracentrifugation was not able to separate nuclei from myofibrils using cross-linked muscle tissue, we devised a sequential filtration procedure to obtain high-quality nuclei devoid of significant myofibril contamination. We subsequently prepared chromatin by using an ultrasonicator, and ChIP assays with anti-BMAL1 antibody revealed robust circadian binding pattern of BMAL1 to target gene promoters. This filtration protocol constitutes an easily applicable method to isolate high-quality nuclei from cross-linked skeletal muscle tissue, allowing consistent sample processing for circadian and other time-sensitive studies. In combination with next-generation sequencing (NGS), our method can be deployed for various mechanistic and genomic studies focusing on skeletal muscle function.

  19. Electrically-receptive and thermally-responsive paper-based sensor chip for rapid detection of bacterial cells.

    PubMed

    Khan, Muhammad S; Misra, Santosh K; Dighe, Ketan; Wang, Zhen; Schwartz-Duval, Aaron S; Sar, Dinabandhu; Pan, Dipanjan

    2018-07-01

    Although significant technological advancements have been made in the development of analytical biosensor chips for detecting bacterial strains (E. coli, S. Mutans and B. Subtilis), critical requirements i.e. limit of detection (LOD), fast time of response, ultra-sensitivity with high reproducibility and good shelf-life with robust sensing capability have yet to be met within a single sensor chip. In order to achieve these criteria, we present an electrically-receptive thermally-responsive (ER-TR) sensor chip comprised of simple filter paper used as substrate coated with composite of poly(N-isopropylacrylamide) polymer (PNIPAm) - graphene nanoplatelet (GR) followed by evaporation of Au electrodes for capturing both Gram-positive (S. mutans and B. subtilis) and Gram-negative (E. coli) bacterial cells in real-time. Autoclave water, tap water, lake water and milk samples were tested with ER-TR chip with and without bacterial strains at varying concentration range 10 1 -10 5 cells/mL. The sensor was integrated with in-house built printed circuit board (PCB) to transmit/receive electrical signals. The interaction of E. coli, S. mutans and B. subtilis cells with fibers of PNIPAm-GR resulted in a change of electrical resistance and the readout was monitored wirelessly in real-time using MATLAB algorithm. Finally, prepared ER-TR chip exhibited the reproducibility of 85-97% with shelf-life of up to four weeks after testing with lake water sample. Copyright © 2018 Elsevier B.V. All rights reserved.

  20. Concentration and purification of HIV-1 virions by microfluidic separation of superparamagnetic nanoparticles

    PubMed Central

    Chen, Grace Dongqing; Alberts, Catharina Johanna

    2009-01-01

    The low concentration and complex sample matrix of many clinical and environmental viral samples presents a significant challenge in the development of low cost, point-of-care viral assays. To address this problem, we investigated the use of a microfluidic passive magnetic separator combined with on-chip mixer to both purify and concentrate whole particle HIV-1 virions. Virus-containing plasma samples are first mixed to allow specific binding of the viral particles with antibody-conjugated superparamagnetic nanoparticles, and several passive mixer geometries were assessed for their mixing efficiencies. The virus-nanoparticle complexes are then separated from the plasma in a novel magnetic separation chamber, where packed micron-sized ferromagnetic particles serve as high magnetic gradient concentrators for an externally applied magnetic field. Thereafter, a viral lysis buffer was flowed through the chip and the released HIV proteins were assayed off-chip. Viral protein extraction efficiencies of 62% and 45% were achieved at 10uL/min and 30uL/min throughputs respectively. More importantly, an 80-fold concentration was observed for an initial sample volume of 1mL, and a 44-fold concentration for an initial sample volume of 0.5mL. The system is broadly applicable to microscale sample preparation of any viral sample and can be used for nucleic acid extraction as well as 40–80 fold enrichment of target viruses. PMID:19954210

  1. Magnetic particles for in vitro molecular diagnosis: From sample preparation to integration into microsystems.

    PubMed

    Tangchaikeeree, Tienrat; Polpanich, Duangporn; Elaissari, Abdelhamid; Jangpatarapongsa, Kulachart

    2017-10-01

    Colloidal magnetic particles (MPs) have been developed in association with molecular diagnosis for several decades. MPs have the great advantage of easy manipulation using a magnet. In nucleic acid detection, these particles can act as a capture support for rapid and simple biomolecule separation. The surfaces of MPs can be modified by coating with various polymer materials to provide functionalization for different applications. The use of MPs enhances the sensitivity and specificity of detection due to the specific activity on the surface of the particles. Practical applications of MPs demonstrate greater efficiency than conventional methods. Beyond traditional detection, MPs have been successfully adopted as a smart carrier in microfluidic and lab-on-a-chip biosensors. The versatility of MPs has enabled their integration into small single detection units. MPs-based biosensors can facilitate rapid and highly sensitive detection of very small amounts of a sample. In this review, the application of MPs to the detection of nucleic acids, from sample preparation to analytical readout systems, is described. State-of-the-art integrated microsystems containing microfluidic and lab-on-a-chip biosensors for the nucleic acid detection are also addressed. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Preparation Characterization and Electrical Study of New Polymeric Mixture (Consist of Three Polymers) Nanocomposites

    NASA Astrophysics Data System (ADS)

    AL-Abodi, Entisar E.; Farouk, Azhar

    2018-05-01

    By using hummers’ method, graphene oxide (GO) was synthesized and by reducing it gave reduced graphene oxide (RGO). The polymeric blend contain three polymers; Poly Aniline (PANI), Poly Vinyl Acetate( PVAc),and Pecten(Pc) wich have been prepared at studied amount. The composites for above polymers with various concentrations of, graphene oxide (GO) and with reduced graphene oxide (RGO)were prepared, and than pour into films(chips). The dielectric constant properties of chips were measured, which its point the electrical conductivity values for the prepared chips increase with increasing of frequency. As well, the electrical conductivity is research in terms of the Arrhenius plot, it is plotted against the reverse temperature for the prepared films at different applied frequencies.

  3. m-Cresol purple functionalized surface enhanced Raman scattering paper chips for highly sensitive detection of pH in the neutral pH range.

    PubMed

    Zou, Xinxin; Wang, Yunqing; Liu, Wanhui; Chen, Lingxin

    2017-06-26

    Herein, a pH sensitive paper SERS chip was prepared by selecting m-cresol purple, a molecule with halochromic properties in the neutral pH range as a Raman reporter. The adsorbed m-cresol purple underwent a reversible change in its electronic configuration from a non-resonant species to a resonant species, which resulted in a significant Raman signal intensity variation due to the transformation of the sensing mode from SERS to surface-enhanced resonance Raman scattering (SERRS). The chips have a sensitive pH range of 6.0 to 8.0 and exhibited good performance for the detection of natural water samples with detection precision of approximately 0.03 pH units, suggesting great potential for environmental pH monitoring applications.

  4. A lab-on-chip for malaria diagnosis and surveillance

    PubMed Central

    2014-01-01

    Background Access to timely and accurate diagnostic tests has a significant impact in the management of diseases of global concern such as malaria. While molecular diagnostics satisfy this need effectively in developed countries, barriers in technology, reagent storage, cost and expertise have hampered the introduction of these methods in developing countries. In this study a simple, lab-on-chip PCR diagnostic was created for malaria that overcomes these challenges. Methods The platform consists of a disposable plastic chip and a low-cost, portable, real-time PCR machine. The chip contains a desiccated hydrogel with reagents needed for Plasmodium specific PCR. Chips can be stored at room temperature and used on demand by rehydrating the gel with unprocessed blood, avoiding the need for sample preparation. These chips were run on a custom-built instrument containing a Peltier element for thermal cycling and a laser/camera setup for amplicon detection. Results This diagnostic was capable of detecting all Plasmodium species with a limit of detection for Plasmodium falciparum of 2 parasites/μL of blood. This exceeds the sensitivity of microscopy, the current standard for diagnosis in the field, by ten to fifty-fold. In a blind panel of 188 patient samples from a hyper-endemic region of malaria transmission in Uganda, the diagnostic had high sensitivity (97.4%) and specificity (93.8%) versus conventional real-time PCR. The test also distinguished the two most prevalent malaria species in mixed infections, P. falciparum and Plasmodium vivax. A second blind panel of 38 patient samples was tested on a streamlined instrument with LED-based excitation, achieving a sensitivity of 96.7% and a specificity of 100%. Conclusions These results describe the development of a lab-on-chip PCR diagnostic from initial concept to ready-for-manufacture design. This platform will be useful in front-line malaria diagnosis, elimination programmes, and clinical trials. Furthermore, test chips can be adapted to detect other pathogens for a differential diagnosis in the field. The flexibility, reliability, and robustness of this technology hold much promise for its use as a novel molecular diagnostic platform in developing countries. PMID:24885206

  5. Integrated circuit package with lead structure and method of preparing the same

    NASA Technical Reports Server (NTRS)

    Kennedy, B. W. (Inventor)

    1973-01-01

    A beam-lead integrated circuit package assembly including a beam-lead integrated circuit chip, a lead frame array bonded to projecting fingers of the chip, a rubber potting compound disposed around the chip, and an encapsulating molded plastic is described. The lead frame array is prepared by photographically printing a lead pattern on a base metal sheet, selectively etching to remove metal between leads, and plating with gold. Joining of the chip to the lead frame array is carried out by thermocompression bonding of mating goldplated surfaces. A small amount of silicone rubber is then applied to cover the chip and bonded joints, and the package is encapsulated with epoxy resin, applied by molding.

  6. The dual role of deposited microbead plug (DMBP): a blood filter and a conjugate reagent carrier toward point-of-care microfluidic immunoassay.

    PubMed

    Li, Chunyu; Liu, Chong; Xu, Zheng; Li, Jingmin

    2012-08-15

    To set up a point-of-care whole-blood immunoassay system, sample preparation and on-chip storage of conjugate reagents are indispensable functional units. Here, we merge these functions into a deposited microbead plug (DMBP) to simultaneously play the roles of a blood filter and a conjugate reagent carrier. The DMBP was easily fabricated by the use of natural deposition of beads without the need of weirs. Conjugate reagents (FITC labeled antibodies used here) were incorporated into the DMBP during the assembly of the DMBP. To demonstrate the ability of the DMBP, we constructed a DMBP-based microfluidic chip and used it for the detection of human IgG (hIgG). The DMBP enabled to remove blood cells from whole blood and provide the pure plasma for the downstream on-chip immunoreactions. The release of reconstituted FITC labeled antibodies from the DMBP was controlled in a passive fashion. Dry FITC labeled antibodies retained at least 81% of their activity after 60 days of storage at the room temperature. The DMBP presented here makes an important step towards the development of the self-contained, integrated, sample-to-answer microfluidic chips for point-of-care diagnostics. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. On-chip sample pretreatment using a porous polymer monolithic column for solid-phase microextraction and chemiluminescence determination of catechins in green tea.

    PubMed

    Lin, Ling; Chen, Hui; Wei, Huibin; Wang, Feng; Lin, Jin-Ming

    2011-10-21

    A porous polymer monolithic column for solid-phase microextraction and chemiluminescence detection was integrated into a simple microfluidic chip for the extraction and determination of catechins in green tea. The porous polymer was prepared by poly(glycidyl methacrylate-co-ethylene dimethacrylate) and modified with ethylenediamine. Catechins can be concentrated in the porous polymer monolithic column and react with potassium permanganate to give chemiluminescence. The microfluidic chip is reusable with high sensitivity and very low reagent consumption. The on-line preconcentration and detection can be realized without an elution step. The enrichment factor was calculated to be about 20 for catechins. The relative chemiluminescence intensity increased linearly with concentration of catechin from 5.0 × 10(-9) to 1.0 × 10(-6) M and the limit of detection was 1.0 × 10(-9) M. The proposed method was applied to determine catechin in green tea. The recoveries are from 90% to 110% which benefits the actual application for green tea samples.

  8. Applications of Blue Light-curing Acrylic Resin to Forensic Sample Preparation and Microtomy.

    PubMed

    Groves, Ethan; Palenik, Christopher S

    2016-03-01

    This study discusses the results of an evaluation of a one-part blue light-curing acrylic resin for embedding trace evidence prior to the preparation of thin sections with a microtome. Through a comparison to several epoxy resins, the physical properties relevant to both trace evidence examination and analytical microscopy in general, including as viscosity, clarity, color, hardness, and cure speed, were explored. Finally, thin sections from paint samples embedded in this acrylic resin were evaluated to determine if, through smearing or impregnation, the resin contributed to the infrared spectra. The results of this study show that blue light-curing acrylic resins provide the desired properties of an embedding medium, generate high-quality thin sections, and can significantly simplify the preparation of paint chips, fibers and a multitude of other types of microscopic samples in the forensic trace evidence laboratory. © 2015 American Academy of Forensic Sciences.

  9. Production of conjugated linoleic acid-rich potato chips.

    PubMed

    Jain, Vishal P; Proctor, Andrew

    2007-01-01

    Conjugated linoleic acid (CLA) is found primarily in diary and beef products, but the health benefits of CLA can only be realized if they are consumed at much greater levels than a normal healthy dietary intake. We have recently shown that a CLA-rich soy oil can be produced by simple isomerization of linoleic acid in soy oil by photoirradiation. This oil may allow greatly increased dietary CLA without significantly elevating fat intake. The objective of this study was to prepare CLA-rich potato chips by frying in CLA-rich soy oil. Soy oil was photoisomerized in the presence of iodine catalyst with UV/visible light. The irradiated oil was clay processed to remove the residual iodine and this oil was then used to fry potato chips. Oil was extracted from fried chips and analyzed for its CLA content with gas chromatography. A 1-oz serving of CLA-rich potato chips contained approximately 2.4 g CLA as compared to 0.1 g CLA in 3-oz serving of steak fillet and 0.06 g CLA in 8-oz serving of whole milk. The peroxide value of the oil extracted from potato chips was found to be 1 meq/1000 g sample, which was within the acceptable commercial standards. This study may lead to the commercialization of CLA-rich food products.

  10. Microfluidic device for a rapid immobilization of zebrafish larvae in environmental scanning electron microscopy.

    PubMed

    Akagi, Jin; Zhu, Feng; Skommer, Joanna; Hall, Chris J; Crosier, Philip S; Cialkowski, Michal; Wlodkowic, Donald

    2015-03-01

    Small vertebrate model organisms have recently gained popularity as attractive experimental models that enhance our understanding of human tissue and organ development. Despite a large body of evidence using optical spectroscopy for the characterization of small model organism on chip-based devices, no attempts have been so far made to interface microfabricated technologies with environmental scanning electron microscopy (ESEM). Conventional scanning electron microscopy requires high vacuum environments and biological samples must be, therefore, submitted to many preparative procedures to dehydrate, fix, and subsequently stain the sample with gold-palladium deposition. This process is inherently low-throughput and can introduce many analytical artifacts. This work describes a proof-of-concept microfluidic chip-based system for immobilizing zebrafish larvae for ESEM imaging that is performed in a gaseous atmosphere, under low vacuum mode and without any need for sample staining protocols. The microfabricated technology provides a user-friendly and simple interface to perform ESEM imaging on zebrafish larvae. Presented lab-on-a-chip device was fabricated using a high-speed infrared laser micromachining in a biocompatible poly(methyl methacrylate) thermoplastic. It consisted of a reservoir with multiple semispherical microwells designed to hold the yolk of dechorionated zebrafish larvae. Immobilization of the larvae was achieved by a gentle suction generated during blotting of the medium. Trapping region allowed for multiple specimens to be conveniently positioned on the chip-based device within few minutes for ESEM imaging. © 2014 International Society for Advancement of Cytometry.

  11. A Simple and Selective Fluorescent Sensor Chip for Indole-3-Butyric Acid in Mung Bean Sprouts Based on Molecularly Imprinted Polymer Coatings

    PubMed Central

    Chang, Jiahua; Bahethan, Bota; Muhammad, Turghun; Yakup, Burabiye; Abbas, Mamatimin

    2017-01-01

    In this paper, we report the preparation of molecularly imprinted polymer coatings on quartz chips for selective solid-phase microextraction and fluorescence sensing of the auxin, indole-3-butyric acid. The multiple copolymerization method was used to prepare polymer coatings on silylated quartz chips. The polymer preparation conditions (e.g., the solvent, monomer, and cross-linker) were investigated systemically to enhance the binding performance of the imprinted coatings. Direct solid-phase fluorescence measurements on the chips facilitated monitoring changes in coating performance. The average binding capacity of an imprinted polymer coated chip was approximately 152.9 µg, which was higher than that of a non-imprinted polymer coated chip (60.8 µg); the imprinted coatings showed the highest binding to IBA among the structural analogues, indicating that the coatings possess high selectivity toward the template molecule. The developed method was used for the determination of the auxin in mung bean extraction, and the recovery was found to be in the range of 91.5% to 97.5%, with an RSD (n = 3) of less than 7.4%. Thus, the present study provides a simple method for fabricating a fluorescent sensor chip for selective analysis. PMID:28837081

  12. Microchannel gel electrophoretic separation systems and methods for preparing and using

    DOEpatents

    Herr, Amy E; Singh, Anup K; Throckmorton, Daniel J

    2015-02-24

    A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays that are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platform enables integration, parallel assays, automation and development of portable devices.

  13. Droplet-based Biosensing for Lab-on-a-Chip, Open Microfluidics Platforms

    PubMed Central

    Dak, Piyush; Ebrahimi, Aida; Swaminathan, Vikhram; Duarte-Guevara, Carlos; Bashir, Rashid; Alam, Muhammad A.

    2016-01-01

    Low cost, portable sensors can transform health care by bringing easily available diagnostic devices to low and middle income population, particularly in developing countries. Sample preparation, analyte handling and labeling are primary cost concerns for traditional lab-based diagnostic systems. Lab-on-a-chip (LoC) platforms based on droplet-based microfluidics promise to integrate and automate these complex and expensive laboratory procedures onto a single chip; the cost will be further reduced if label-free biosensors could be integrated onto the LoC platforms. Here, we review some recent developments of label-free, droplet-based biosensors, compatible with “open” digital microfluidic systems. These low-cost droplet-based biosensors overcome some of the fundamental limitations of the classical sensors, enabling timely diagnosis. We identify the key challenges that must be addressed to make these sensors commercially viable and summarize a number of promising research directions. PMID:27089377

  14. Microchannel gel electrophoretic separation systems and methods for preparing and using

    DOEpatents

    Herr, Amy; Singh, Anup K; Throckmorton, Daniel J

    2013-09-03

    A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays that are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platform enables integration, parallel assays, automation and development of portable devices.

  15. Carbon nanotubes for voltage reduction and throughput enhancement of electrical cell lysis on a lab-on-a-chip.

    PubMed

    Shahini, Mehdi; Yeow, John T W

    2011-08-12

    We report on the enhancement of electrical cell lysis using carbon nanotubes (CNTs). Electrical cell lysis systems are widely utilized in microchips as they are well suited to integration into lab-on-a-chip devices. However, cell lysis based on electrical mechanisms has high voltage requirements. Here, we demonstrate that by incorporating CNTs into microfluidic electrolysis systems, the required voltage for lysis is reduced by half and the lysis throughput at low voltages is improved by ten times, compared to non-CNT microchips. In our experiment, E. coli cells are lysed while passing through an electric field in a microchannel. Based on the lightning rod effect, the electric field strengthened at the tip of the CNTs enhances cell lysis at lower voltage and higher throughput. This approach enables easy integration of cell lysis with other on-chip high-throughput sample-preparation processes.

  16. Droplet-based Biosensing for Lab-on-a-Chip, Open Microfluidics Platforms.

    PubMed

    Dak, Piyush; Ebrahimi, Aida; Swaminathan, Vikhram; Duarte-Guevara, Carlos; Bashir, Rashid; Alam, Muhammad A

    2016-04-14

    Low cost, portable sensors can transform health care by bringing easily available diagnostic devices to low and middle income population, particularly in developing countries. Sample preparation, analyte handling and labeling are primary cost concerns for traditional lab-based diagnostic systems. Lab-on-a-chip (LoC) platforms based on droplet-based microfluidics promise to integrate and automate these complex and expensive laboratory procedures onto a single chip; the cost will be further reduced if label-free biosensors could be integrated onto the LoC platforms. Here, we review some recent developments of label-free, droplet-based biosensors, compatible with "open" digital microfluidic systems. These low-cost droplet-based biosensors overcome some of the fundamental limitations of the classical sensors, enabling timely diagnosis. We identify the key challenges that must be addressed to make these sensors commercially viable and summarize a number of promising research directions.

  17. Microfluidic free-flow zone electrophoresis and isotachophoresis using carbon black nano-composite PDMS sidewall membranes.

    PubMed

    Fu, Xiaotong; Mavrogiannis, Nicholas; Ibo, Markela; Crivellari, Francesca; Gagnon, Zachary R

    2017-01-01

    We present a new type of free-flow electrophoresis (FFE) device for performing on-chip microfluidic isotachophoresis and zone electrophoresis. FFE is performed using metal gallium electrodes, which are isolated from a main microfluidic flow channel using thin micron-scale polydimethylsiloxane/carbon black (PDMS/CB) composite membranes integrated directly into the sidewalls of the microfluidic channel. The thin membrane allows for field penetration and effective electrophoresis, but serves to prevent bubble generation at the electrodes from electrolysis. We experimentally demonstrate the ability to use this platform to perform on-chip free-flow electrophoretic separation and isotachophoretic concentration. Due to the small size and simple fabrication procedure, this PDMS/CB platform could be used as a part of an on-chip upstream sample preparation toolkit for portable microfluidic diagnostic applications. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Microfluidic Sample Preparation for Diagnostic Cytopathology

    PubMed Central

    Mach, Albert J.; Adeyiga, Oladunni B.; Di Carlo, Dino

    2014-01-01

    The cellular components of body fluids are routinely analyzed to identify disease and treatment approaches. While significant focus has been placed on developing cell analysis technologies, tools to automate the preparation of cellular specimens have been more limited, especially for body fluids beyond blood. Preparation steps include separating, concentrating, and exposing cells to reagents. Sample preparation continues to be routinely performed off-chip by technicians, preventing cell-based point-of-care diagnostics, increasing the cost of tests, and reducing the consistency of the final analysis following multiple manually-performed steps. Here, we review the assortment of biofluids for which suspended cells are analyzed, along with their characteristics and diagnostic value. We present an overview of the conventional sample preparation processes for cytological diagnosis. We finally discuss the challenges and opportunities in developing microfluidic devices for the purpose of automating or miniaturizing these processes, with particular emphases on preparing large or small volume samples, working with samples of high cellularity, automating multi-step processes, and obtaining high purity subpopulations of cells. We hope to convey the importance of and help identify new research directions addressing the vast biological and clinical applications in preparing and analyzing the array of available biological fluids. Successfully addressing the challenges described in this review can lead to inexpensive systems to improve diagnostic accuracy while simultaneously reducing overall systemic healthcare costs. PMID:23380972

  19. A nano-patterned self assembled monolayer (SAM) rutile titania cancer chip for rapid, low cost, highly sensitive, direct cancer analysis in MALDI-MS.

    PubMed

    Manikandan, M; Gopal, Judy; Hasan, Nazim; Wu, Hui-Fen

    2014-12-01

    We developed a cancer chip by nano-patterning a highly sensitive SAM titanium surface capable of capturing and sensing concentrations as low as 10 cancer cells/mL from the environment by Matrix Assisted Laser Desorption and Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS). The current approach evades any form of pretreatment and sample preparation processes; it is time saving and does not require the (expensive) conventional MALDI target plate. The home made aluminium (Al) target holder cost, on which we loaded the cancer chips for MALDI-TOF MS analysis, is about 60 USD. While the conventional stainless steel MALDI target plate is more than 700 USD. The SAM surface was an effective platform leading to on-chip direct MALDI-MS detection of cancer cells. We compared the functionality of this chip with the unmodified titanium surfaces and thermally oxidized (TO) titanium surfaces. The lowest detectable concentration of the TO chip was 10(3) cells/mL, while the lowest detectable concentration of the control or unmodified titanium chips was 10(6) cells/mL. Compared to the control surface, the SAM cancer chip showed 100,000 times of enhanced sensitivity and compared with the TO chip, 1000 times of increased sensitivity. The high sensitivity of the SAM surfaces is attributed to the presence of the rutile SAM, surface roughness and surface wettability as confirmed by AFM, XRD, contact angle microscope and FE-SEM. This study opens a new avenue for the potent application of the SAM cancer chip for direct cancer diagnosis by MALDI-TOF MS in the near future. Copyright © 2014. Published by Elsevier B.V.

  20. Integrated DNA and RNA extraction and purification on an automated microfluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections.

    PubMed

    Van Heirstraeten, Liesbet; Spang, Peter; Schwind, Carmen; Drese, Klaus S; Ritzi-Lehnert, Marion; Nieto, Benjamin; Camps, Marta; Landgraf, Bryan; Guasch, Francesc; Corbera, Antoni Homs; Samitier, Josep; Goossens, Herman; Malhotra-Kumar, Surbhi; Roeser, Tina

    2014-05-07

    In this paper, we describe the development of an automated sample preparation procedure for etiological agents of community-acquired lower respiratory tract infections (CA-LRTI). The consecutive assay steps, including sample re-suspension, pre-treatment, lysis, nucleic acid purification, and concentration, were integrated into a microfluidic lab-on-a-chip (LOC) cassette that is operated hands-free by a demonstrator setup, providing fluidic and valve actuation. The performance of the assay was evaluated on viral and Gram-positive and Gram-negative bacterial broth cultures previously sampled using a nasopharyngeal swab. Sample preparation on the microfluidic cassette resulted in higher or similar concentrations of pure bacterial DNA or viral RNA compared to manual benchtop experiments. The miniaturization and integration of the complete sample preparation procedure, to extract purified nucleic acids from real samples of CA-LRTI pathogens to, and above, lab quality and efficiency, represent important steps towards its application in a point-of-care test (POCT) for rapid diagnosis of CA-LRTI.

  1. Multiplexed capillary microfluidic immunoassay with smartphone data acquisition for parallel mycotoxin detection.

    PubMed

    Machado, Jessica M D; Soares, Ruben R G; Chu, Virginia; Conde, João P

    2018-01-15

    The field of microfluidics holds great promise for the development of simple and portable lab-on-a-chip systems. The use of capillarity as a means of fluidic manipulation in lab-on-a-chip systems can potentially reduce the complexity of the instrumentation and allow the development of user-friendly devices for point-of-need analyses. In this work, a PDMS microchannel-based, colorimetric, autonomous capillary chip provides a multiplexed and semi-quantitative immunodetection assay. Results are acquired using a standard smartphone camera and analyzed with a simple gray scale quantification procedure. The performance of this device was tested for the simultaneous detection of the mycotoxins ochratoxin A (OTA), aflatoxin B1 (AFB1) and deoxynivalenol (DON) which are strictly regulated food contaminants with severe detrimental effects on human and animal health. The multiplexed assay was performed approximately within 10min and the achieved sensitivities of<40, 0.1-0.2 and<10ng/mL for OTA, AFB1 and DON, respectively, fall within the majority of currently enforced regulatory and/or recommended limits. Furthermore, to assess the potential of the device to analyze real samples, the immunoassay was successfully validated for these 3 mycotoxins in a corn-based feed sample after a simple sample preparation procedure. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Application of handheld and portable spectrometers for screening acrylamide content in commercial potato chips.

    PubMed

    Ayvaz, Huseyin; Rodriguez-Saona, Luis E

    2015-05-01

    The most common methods for acrylamide analysis in foods require the use of LC-MS/MS and GC-MS. Although these methods have great analytical performance, they need intensive sample preparation, highly specialised instrumentation, and are time consuming. In this study, portable and handheld infrared spectrometers were evaluated as rapid methods for screening acrylamide in potato chips and their performances were compared to those of benchtop infrared systems. The acrylamide content of 64 commercial potato chips (169-2453 μg/kg) was determined by LC-MS/MS. Spectral data were collected using mid-infrared (MIR) and near-infrared (NIR) spectrometers. Partial least squares regression (PLSR) calibration models were developed to predict acrylamide levels. Overall, good linear correlation was found between the predicted acrylamide levels and actual measured acrylamide concentrations by LC-MS/MS (rPred > 0.90 and SEP < 100 μg/kg). Our results indicate that portable and handheld spectrometers can be used as simple and rapid alternatives for acrylamide analysis in potato chips. Copyright © 2014 Elsevier Ltd. All rights reserved.

  3. Geochemistry of CI chondrites: Major and trace elements, and Cu and Zn Isotopes

    NASA Astrophysics Data System (ADS)

    Barrat, J. A.; Zanda, B.; Moynier, F.; Bollinger, C.; Liorzou, C.; Bayon, G.

    2012-04-01

    In order to check the heterogeneity of the CI chondrites and determine the average composition of this group of meteorites, we analyzed a series of six large chips (weighing between 0.6 and 1.2 g) of Orgueil prepared from five different stones. In addition, one sample from each of Ivuna and Alais was analyzed. Although the sizes of the chips used in this study were “large”, our results show evidence for minor chemical heterogeneity in Orgueil, particularly for alkali elements and U. After removal of one outlier sample, the spread of the results is considerably reduced. For most of the 46 elements analyzed in this study, the average composition calculated for Orgueil is in very good agreement with previous CI estimates. This average, obtained with a “large” mass of samples, is analytically homogeneous and is suitable for normalization purposes. Finally, the Cu and Zn isotopic ratios are homogeneously distributed within the CI parent body with a spread of less than 100 ppm per atomic mass unit (amu).

  4. Chip-Based Sensors for Disease Diagnosis

    NASA Astrophysics Data System (ADS)

    Fang, Zhichao

    Nucleic acid analysis is one of the most important disease diagnostic approaches in medical practice, and has been commonly used in cancer biomarker detection, bacterial speciation and many other fields in laboratory. Currently, the application of powerful research methods for genetic analysis, including the polymerase chain reaction (PCR), DNA sequencing, and gene expression profiling using fluorescence microarrays, are not widely used in hospitals and extended-care units due to high-cost, long detection times, and extensive sample preparation. Bioassays, especially chip-based electrochemical sensors, may be suitable for the next generation of rapid, sensitive, and multiplexed detection tools. Herein, we report three different microelectrode platforms with capabilities enabled by nano- and microtechnology: nanoelectrode ensembles (NEEs), nanostructured microelectrodes (NMEs), and hierarchical nanostructured microelectrodes (HNMEs), all of which are able to directly detect unpurified RNA in clinical samples without enzymatic amplification. Biomarkers that are cancer and infectious disease relevant to clinical medicine were chosen to be the targets. Markers were successfully detected with clinically-relevant sensitivity. Using peptide nucleic acids (PNAs) as probes and an electrocatalytic reporter system, NEEs were able to detect prostate cancer-related gene fusions in tumor tissue samples with 100 ng of RNA. The development of NMEs improved the sensitivity of the assay further to 10 aM of DNA target, and multiplexed detection of RNA sequences of different prostate cancer-related gene fusion types was achieved on the chip-based NMEs platform. An HNMEs chip integrated with a bacterial lysis device was able to detect as few as 25 cfu bacteria in 30 minutes and monitor the detection in real time. Bacterial detection could also be performed in neat urine samples. The development of these versatile clinical diagnostic tools could be extended to the detection of various cancers, genetic, and infectious diseases.

  5. [Lab-on-a-chip systems in the point-of-care diagnostics].

    PubMed

    Szabó, Barnabás; Borbíró, András; Fürjes, Péter

    2015-12-27

    The need in modern medicine for near-patient diagnostics being able to accelerate therapeutic decisions and possibly replacing laboratory measurements is significantly growing. Reliable and cost-effective bioanalytical measurement systems are required which - acting as a micro-laboratory - contain integrated biomolecular recognition, sensing, signal processing and complex microfluidic sample preparation modules. These micro- and nanofabricated Lab-on-a-chip systems open new perspectives in the diagnostic supply chain, since they are able even for quantitative, high-precision and immediate analysis of special disease specific molecular markers or their combinations from a single drop of sample. Accordingly, crucial requirements regarding the instruments and the analytical methods are the high selectivity, extremely low detection limit, short response time and integrability into the healthcare information networks. All these features can make the hierarchical examination chain shorten, and revolutionize laboratory diagnostics, evolving a brand new situation in therapeutic intervention.

  6. Microfluidic Remote Loading for Rapid Single-Step Liposomal Drug Preparation

    PubMed Central

    Hood, R.R.; Vreeland, W. N.; DeVoe, D.L.

    2014-01-01

    Microfluidic-directed formation of liposomes is combined with in-line sample purification and remote drug loading for single step, continuous-flow synthesis of nanoscale vesicles containing high concentrations of stably loaded drug compounds. Using an on-chip microdialysis element, the system enables rapid formation of large transmembrane pH and ion gradients, followed by immediate introduction of amphipathic drug for real-time remote loading into the liposomes. The microfluidic process enables in-line formation of drug-laden liposomes with drug:lipid molar ratios of up to 1.3, and a total on-chip residence time of approximately 3 min, representing a significant improvement over conventional bulk-scale methods which require hours to days for combined liposome synthesis and remote drug loading. The microfluidic platform may be further optimized to support real-time generation of purified liposomal drug formulations with high concentrations of drugs and minimal reagent waste for effective liposomal drug preparation at or near the point of care. PMID:25003823

  7. Submicron magnetic core conducting polypyrrole polymer shell: Preparation and characterization.

    PubMed

    Tenório-Neto, Ernandes Taveira; Baraket, Abdoullatif; Kabbaj, Dounia; Zine, Nadia; Errachid, Abdelhamid; Fessi, Hatem; Kunita, Marcos Hiroiuqui; Elaissari, Abdelhamid

    2016-04-01

    Magnetic particles are of great interest in various biomedical applications, such as, sample preparation, in vitro biomedical diagnosis, and both in vivo diagnosis and therapy. For in vitro applications and especially in labs-on-a-chip, microfluidics, microsystems, or biosensors, the needed magnetic dispersion should answer various criteria, for instance, submicron size in order to avoid a rapid sedimentation rate, fast separations under an applied magnetic field, and appreciable colloidal stability (stable dispersion under shearing process). Then, the aim of this work was to prepare highly magnetic particles with a magnetic core and conducting polymer shell particles in order to be used not only as a carrier, but also for the in vitro detection step. The prepared magnetic seed dispersions were functionalized using pyrrole and pyrrole-2-carboxylic acid. The obtained core-shell particles were characterized in terms of particle size, size distribution, magnetization properties, FTIR analysis, surface morphology, chemical composition, and finally, the conducting property of those particles were evaluated by cyclic voltammetry. The obtained functional submicron highly magnetic particles are found to be conducting material bearing function carboxylic group on the surface. These promising conducting magnetic particles can be used for both transport and lab-on-a-chip detection. Copyright © 2015. Published by Elsevier B.V.

  8. Effect of cooking method (baking compared with frying) on acrylamide level of potato chips.

    PubMed

    Palazoğlu, T Koray; Savran, Derya; Gökmen, Vural

    2010-01-01

    The effect of cooking method (baking compared with frying) on acrylamide level of potato chips was investigated in this study. Baking and frying experiments were conducted at 170, 180, and 190 degrees C using potato slices with a thickness of 1.4 mm. Raw potatoes were analyzed for reducing sugars and asparagine. Surface and internal temperatures of potato slices were monitored during the experiments to better explain the results. Fried and baked chips were analyzed for acrylamide content using an LC-MS method. The results showed that acrylamide level of potato chips prepared by frying increased with frying temperature (19.6 ng/g at 170 degrees C, 39 ng/g at 180 degrees C, and 95 ng/g at 190 degrees C). In baking, however, the highest acrylamide level was observed in potato chips prepared at 170 degrees C (47.8 ng/g at 170 degrees C, 19.3 ng/g at 180 degrees C, and 29.7 ng/g at 190 degrees C). The results showed that baking at 170 degrees C more than doubled the acrylamide amount that formed upon frying at the same temperature, whereas at 180 and 190 degrees C, the acrylamide levels of chips prepared by baking were lower than their fried counterparts.

  9. Microfluidic chip based nano liquid chromatography coupled to tandem mass spectrometry for the determination of abused drugs and metabolites in human hair.

    PubMed

    Zhu, Kevin Y; Leung, K Wing; Ting, Annie K L; Wong, Zack C F; Ng, Winki Y Y; Choi, Roy C Y; Dong, Tina T X; Wang, Tiejie; Lau, David T W; Tsim, Karl W K

    2012-03-01

    A microfluidic chip based nano-HPLC coupled to tandem mass spectrometry (nano-HPLC-Chip-MS/MS) has been developed for simultaneous measurement of abused drugs and metabolites: cocaine, benzoylecgonine, cocaethylene, norcocaine, morphine, codeine, 6-acetylmorphine, phencyclidine, amphetamine, methamphetamine, MDMA, MDA, MDEA, and methadone in the hair of drug abusers. The microfluidic chip was fabricated by laminating polyimide films and it integrated an enrichment column, an analytical column and a nanospray tip. Drugs were extracted from hairs by sonication, and the chromatographic separation was achieved in 15 min. The drug identification and quantification criteria were fulfilled by the triple quardropule tandem mass spectrometry. The linear regression analysis was calibrated by deuterated internal standards with all of the R(2) at least over 0.993. The limit of detection (LOD) and the limit of quantification (LOQ) were from 0.1 to 0.75 and 0.2 to 1.25 pg/mg, respectively. The validation parameters including selectivity, accuracy, precision, stability, and matrix effect were also evaluated here. In conclusion, the developed sample preparation method coupled with the nano-HPLC-Chip-MS/MS method was able to reveal the presence of drugs in hairs from the drug abusers, with the enhanced sensitivity, compared with the conventional HPLC-MS/MS.

  10. Assessment of Complement Activation by Nanoparticles: Development of a SPR Based Method and Comparison with Current High Throughput Methods.

    PubMed

    Coty, Jean-Baptiste; Noiray, Magali; Vauthier, Christine

    2018-04-26

    A Surface Plasmon Resonance chip (SPR) was developed to study the activation of complement system triggered by nanomaterials in contact with human serum, which is an important concern today to warrant safety of nanomedicines. The developed chip was tested for its specificity in complex medium and its longevity of use. It was then employed to assess the release of complement fragments upon incubation of nanoparticles in serum. A comparison was made with other current methods assessing complement activation (μC-IE, ELISA). The SPR chip was found to give a consistent response for C3a release upon activation by nanoparticles. Results were similar to those obtained by μC-IE. However, ELISA detection of iC3b fragments showed an explained high non-specific background. The impact of sample preparation preceding the analysis was assessed with the newly develop SPR method. The removal of nanoparticles before analysis showed an important modification in the obtained response, possibly leading to false negative results. The SPR chip developed in this work allows for an automated assessment of complement activation triggered by nanoparticles with possibility of multiplexed analysis. The design of the chip proved to give consistent results of complement activation by nanoparticles.

  11. Chip-based nanoflow high performance liquid chromatography coupled to mass spectrometry for profiling of soybean flavonoids.

    PubMed

    Chang, Yuwei; Zhao, Chunxia; Wu, Zeming; Zhou, Jia; Zhao, Sumin; Lu, Xin; Xu, Guowang

    2012-08-01

    In this work a chip-based nano HPLC coupled MS (HPLC-chip/MS) method with a simple sample preparation procedure was developed for the flavonoid profiling of soybean. The analytical properties of the method including the linearity (R(2) , 0.992-0.995), reproducibility (RSD, 1.50-7.66%), intraday precision (RSD, 1.41-5.14%) and interday precision (RSD, 2.76-16.90%) were satisfactory. Compared with the conventional HPLC/MS method, a fast extraction and analysis procedure was applied and more flavonoids were detected in a single run. Additionally, 13 flavonoids in soybean seed were identified for the first time. The method was then applied to the profiling of six varieties of soybean sowed at the same place. A clear discrimination was observed among different cultivars, three isoflavones, accounting for nearly 80% of total flavonoid contents, were found increased in the spring soybeans compared with the summer cultivars. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Chip calorimetry for evaluation of biofilm treatment with biocides, antibiotics, and biological agents.

    PubMed

    Morais, Frida Mariana; Buchholz, Friederike; Maskow, Thomas

    2014-01-01

    Any growth or bioconversion in biofilms is accompanied by the release of heat. The heat (in J) is tightly related to the stoichiometry of the respective process via law of Hess, and the heat production rate (in W or J/s) is additionally related to the process kinetics. This heat and the heat production rate can nowadays be measured by modern calorimetry with extremely high sensitivity. Flow-through calorimetry allows the measurement of bioprocesses in biofilms in real time, without the need of invasive sample preparation and disturbing of biofilm processes. Furthermore, it can be applied for long-term measurements and is even applicable to turbid media. Chip or miniaturized calorimeters have the additional advantages of extremely short thermal equilibration times and the requirement of very small amounts of media and chemicals. The precision of flow-through chip calorimeters (about 3 mW/L) allows the detection of early stages of biofilm development (about 10(5) bacteria cm(-2)).

  13. A disposable, self-contained PCR chip.

    PubMed

    Kim, Jitae; Byun, Doyoung; Mauk, Michael G; Bau, Haim H

    2009-02-21

    A disposable, self-contained polymerase chain reaction (PCR) chip with on-board stored, just-on-time releasable, paraffin-passivated, dry reagents is described. During both storage and sample preparation, the paraffin immobilizes and protects the stored reagents. Fluid flow through the reactor leaves the reagents undisturbed. Prior to the amplification step, the chamber is filled with target analyte suspended in water. Upon heating the PCR chamber to the DNA's denaturation temperature, the paraffin melts and moves out of the way, and the reagents are released and hydrated. To better understand the reagent release process, a scaled up model of the reactor was constructed and the paraffin migration was visualized. Experiments were carried out with a 30 microl reactor demonstrating detectable amplification (with agarose gel electrophoresis) of 10 fg ( approximately 200 copies) of lambda DNA template. The in-reactor storage and on-time release of the PCR reagents reduce the number of needed operations and significantly simplifies the flow control that would, otherwise, be needed in lab-on-chip devices.

  14. A Disposable, Self-Contained PCR Chip

    PubMed Central

    Kim, Jitae; Byun, Doyoung; Mauk, Michael G.; Bau, Haim H.

    2009-01-01

    A disposable, self-contained polymerase chain reaction (PCR) chip with on-board stored, just on time releasable, paraffin-passivated, dry reagents is described. During both storage and sample preparation, the paraffin immobilizes and protects the stored reagents. Fluid flow through the reactor leaves the reagents undisturbed. Prior to the amplification step, the chamber is filled with target analyte suspended in water. Upon heating the PCR chamber to the DNA’s denaturation temperature, the paraffin melts and moves out of the way, and the reagents are released and hydrated. To better understand the reagent release process, a scaled up model of the reactor was constructed and the paraffin migration was visualized. Experiments were carried out with a 30 μl reactor demonstrating detectable amplification (with agarose gel electrophoresis) of 10 fg (~200 copies) of lambda DNA template. The in-reactor storage and on-time release of the PCR reagents reduce the number of needed operations and significantly simplify the flow control that would, otherwise, be needed in lab-on-chip devices. PMID:19190797

  15. Detection of sepsis in patient blood samples using CD64 expression in a microfluidic cell separation device.

    PubMed

    Zhang, Ye; Li, Wenjie; Zhou, Yun; Johnson, Amanda; Venable, Amanda; Hassan, Ahmed; Griswold, John; Pappas, Dimitri

    2017-12-18

    A microfluidic affinity separation device was developed for the detection of sepsis in critical care patients. An affinity capture method was developed to capture cells based on changes in CD64 expression in a single, simple microfluidic chip for sepsis detection. Both sepsis patient samples and a laboratory CD64+ expression model were used to validate the microfluidic assay. Flow cytometry analysis showed that the chip cell capture had a linear relationship with CD64 expression in laboratory models. The Sepsis Chip detected an increase in upregulated neutrophil-like cells when the upregulated cell population is as low as 10% of total cells spiked into commercially available aseptic blood samples. In a proof of concept study, blood samples obtained from sepsis patients within 24 hours of diagnosis were tested on the chip to further validate its performance. On-chip CD64+ cell capture from 10 patient samples (619 ± 340 cells per chip) was significantly different from control samples (32 ± 11 cells per chip) and healthy volunteer samples (228 ± 95 cells per chip). In addition, the on-chip cell capture has a linear relationship with CD64 expression indicating our approach can be used to measure CD64 expression based on total cell capture on Sepsis Chip. Our method has proven to be sensitive, accurate, rapid, and cost-effective. Therefore, this device is a promising detection platform for neutrophil activation and sepsis diagnosis.

  16. Formulation of Saudi Propolis into Biodegradable Chitosan Chips for Vital Pulpotomy.

    PubMed

    Balata, Gihan F; Abdelhady, Mohamed I S; Mahmoud, Ghada M; Matar, Moustafa A; Abd El-Latif, Amani N

    2018-01-01

    Propolis has been widely used to treat oral cavity disorders, such as endodontal and periodontal diseases and microbial infections. The study aimed at the formulation of commercial Saudi propolis into biodegradable chitosan chips and evaluation of its effectiveness as a pulpotomy agent. The standardization of 80% ethanolic propolis extract was performed regarding its total phenolic content, total flavonoid content, quantitative estimation of main polyphenolic constituents and antioxidant activity. Chitosan chips containing propolis extract were prepared by the solvent/ casting method. The investigated variables were % of chitosan polymer (2, 2.5 and 3%), % of plasticizer (1, 5 and 10%) and incorporation of different concentrations of hydroxypropyl methylcellulose (5, 10 and 20% of polymer weight). The chips were characterized for weight and thickness uniformity, content uniformity, pH, percentage moisture loss, swelling index, tensile strength and in vitro propolis release. The optimal propolis chip formulation was further investigated in dogs regarding the short term response of primary dental pulp to propolis chips compared with the most commonly used formocresol preparation. The prepared films were flexible and demonstrated satisfactory physicochemical characteristics. The optimal formulation showed an initial release of about 41.7% of the loaded propolis followed by a sustained release extended up to 7 days. The kinetics study demonstrated that propolis release was controlled by Fick´s diffusion. The optimal propolis chip formulation resulted in less pulpal inflammation compared to formocresol, and produced hard tissue formation in all specimens. Formulation of commercial Saudi propolis as a biodegradable chitosan chip is an effective alternative to the commercially available chemical agents for the treatment of vital pulpotomy. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  17. Chip-based molecularly imprinted monolithic capillary array columns coated GO/SiO2 for selective extraction and sensitive determination of rhodamine B in chili powder.

    PubMed

    Zhai, Haiyun; Huang, Lu; Chen, Zuanguang; Su, Zihao; Yuan, Kaisong; Liang, Guohuan; Pan, Yufang

    2017-01-01

    A novel solid-phase extraction chip embedded with array columns of molecularly imprinted polymer-coated silanized graphene oxide (GO/SiO2-MISPE) was established to detect trace rhodamine B (RB) in chili powder. GO/SiO2-MISPE monolithic columns for RB detection were prepared by optimizing the supporting substrate, template, and polymerizing monomer under mild water bath conditions. Adsorption capacity and specificity, which are critical properties for the application of the GO/SiO2-MISPE monolithic column, were investigated. GO/SiO2-MIP was examined by scanning electron microscopy (SEM) and Fourier transform-infrared spectroscopy. The recovery and the intraday and interday relative standard deviations for RB ranged from 83.7% to 88.4% and 2.5% to 4.0% and the enrichment factors were higher than 110-fold. The chip-based array columns effectively eliminated impurities in chili powder, indicating that the chip-based GO/SiO2-MISPE method was reliable for RB detection in food samples using high-performance liquid chromatography. Accordingly, this method has direct applications for monitoring potentially harmful dyes in processed food. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Physical properties and estimated glycemic index of protein-enriched sorghum based chips.

    PubMed

    Jiang, Hongrui; Hettiararchchy, Navam S; Horax, Ronny

    2018-03-01

    Sorghum is a gluten-free grain and more attention has been given to the nutritional properties and recently its usage as a wheat replacement in food products. In the present work, protein-enriched sorghum based snack chips, prepared from sorghum meal with soy protein isolates and soy flour to meet the final protein content of 35.7%, were produced. The effect of varying baking powder (1.5-2.5%), dough sheet thickness (0.7-1.7 mm), and baking time (6-12 min) on the physical properties of the snack chips was investigated using a central composite design of response surface methodology. Under baking temperature of 160 °C, with baking powder added, the water activity and puffiness of chips significantly increased. Baking time was the most significant factor for all the parameters detected except for puffiness. The optimized conditions of preparing protein-enriched sorghum chips were baking powder 2.5%, dough sheet thickness 0.7 mm, and baking time 7.66 min. The estimated glycemic index (eGI) of the protein-enriched sorghum chips (eGI = 59.8) was significantly lower than soybean-free sorghum chips. The gluten-free protein-enriched sorghum chips developed could be considered as protein rich with lower intermediate-glycemic index classified healthy snacks and potential commercialization.

  19. Fast cholesterol detection using flow injection microfluidic device with functionalized carbon nanotubes based electrochemical sensor.

    PubMed

    Wisitsoraat, A; Sritongkham, P; Karuwan, C; Phokharatkul, D; Maturos, T; Tuantranont, A

    2010-12-15

    This work reports a new cholesterol detection scheme using functionalized carbon nanotube (CNT) electrode in a polydimethylsiloxane/glass based flow injection microfluidic chip. CNTs working, silver reference and platinum counter electrode layers were fabricated on the chip by sputtering and low temperature chemical vapor deposition methods. Cholesterol oxidase prepared in polyvinyl alcohol solution was immobilized on CNTs by in-channel flow technique. Cholesterol analysis based on flow injection chronoamperometric measurement was performed in 150-μm-wide and 150-μm-deep microchannels. Fast and sensitive real-time detection was achieved with high throughput of more than 60 samples per hour and small sample volume of 15 μl. The cholesterol sensor had a linear detection range between 50 and 400 mg/dl. In addition, low cross-sensitivities toward glucose, ascorbic acid, acetaminophen and uric acid were confirmed. The proposed system is promising for clinical diagnostics of cholesterol with high speed real-time detection capability, very low sample consumption, high sensitivity, low interference and good stability. Copyright © 2010 Elsevier B.V. All rights reserved.

  20. TE modes of UV-laser generated waveguides in a planar polymer chip of parabolic refractive index profile

    NASA Astrophysics Data System (ADS)

    Shams El-Din, M. A.

    2018-04-01

    The UV-laser lithographic method is used for the preparation of Polymeric integrated-optical waveguides in a planar polymer chip. The waveguide samples are irradiated by an excimer laser of wavelength 248 nm with different doses and with the same fluencies. The refractive index depth profile for the waveguides, in the first zone is found to have a parabolic shape and Gaussian shape in the second one that can be determined by Mach-Zehnder interferometer. Both the mode field distribution and the effective mode indices for the first zone only are determined by making use of the theoretical mode and the experimental data. It is found that the model field distribution is strongly dependent on the refractive indices for each zone.

  1. Integrated Multi-process Microfluidic Systems for Automating Analysis

    PubMed Central

    Yang, Weichun; Woolley, Adam T.

    2010-01-01

    Microfluidic technologies have been applied extensively in rapid sample analysis. Some current challenges for standard microfluidic systems are relatively high detection limits, and reduced resolving power and peak capacity compared to conventional approaches. The integration of multiple functions and components onto a single platform can overcome these separation and detection limitations of microfluidics. Multiplexed systems can greatly increase peak capacity in multidimensional separations and can increase sample throughput by analyzing many samples simultaneously. On-chip sample preparation, including labeling, preconcentration, cleanup and amplification, can all serve to speed up and automate processes in integrated microfluidic systems. This paper summarizes advances in integrated multi-process microfluidic systems for automated analysis, their benefits and areas for needed improvement. PMID:20514343

  2. Chip-based generation of carbon nanodots via electrochemical oxidation of screen printed carbon electrodes and the applications for efficient cell imaging and electrochemiluminescence enhancement

    NASA Astrophysics Data System (ADS)

    Xu, Yuanhong; Liu, Jingquan; Zhang, Jizhen; Zong, Xidan; Jia, Xiaofang; Li, Dan; Wang, Erkang

    2015-05-01

    A portable lab-on-a-chip methodology to generate ionic liquid-functionalized carbon nanodots (CNDs) was developed via electrochemical oxidation of screen printed carbon electrodes. The CNDs can be successfully applied for efficient cell imaging and solid-state electrochemiluminescence sensor fabrication on the paper-based chips.A portable lab-on-a-chip methodology to generate ionic liquid-functionalized carbon nanodots (CNDs) was developed via electrochemical oxidation of screen printed carbon electrodes. The CNDs can be successfully applied for efficient cell imaging and solid-state electrochemiluminescence sensor fabrication on the paper-based chips. Electronic supplementary information (ESI) available: Experimental section; Fig. S1. XPS spectra of the as-prepared CNDs after being dialyzed for 72 hours; Fig. S2. LSCM images showing time-dependent fluorescence signals of HeLa cells treated by the as-prepared CNDs; Tripropylamine analysis using the Nafion/CNDs modified ECL sensor. See DOI: 10.1039/c5nr01765c

  3. [Advances of the coatings used in columns for capillary electrophoresis and in nanochannels of chips].

    PubMed

    Liu, Chunye; Chen, Jierong

    2005-01-01

    An overview is provided on the advancement and development of coating preparation methodology and materials used in capillaries and channels in microfluidic chip. Discussion is also given on the effects of coatings in the resolutions of separation and the reproducibility of separations. Dynamic coatings and linked coatings, classified as homo-polymers, copolymers and heterocyclic compounds, are further discussed, and so are the methods for the preparation of the coatings by cross-linked reaction, sol-gel process, photomodification and chemical deposition, etc. The discussion will be useful for the optimization of capillary columns that are used in capillary electrophoresis and nanochannels of chip.

  4. The Blue-Chip Graduate: A Four-Year College Plan for Career Success.

    ERIC Educational Resources Information Center

    Osher, Bill; Campbell, Sioux Henley

    A plan is presented that is intended to prepare students to become the type of college graduate (the "blue-chip" graduate) who is successful in a highly competitive job market. The following topics are discussed: "What College Students Want--Job$"; "What Employers Want--Blue-Chip Graduates"; "How To Write a Term Paper, Coordinate Homecoming and…

  5. One-step fabrication of an organ-on-a-chip with spatial heterogeneity using a 3D bioprinting technology.

    PubMed

    Lee, Hyungseok; Cho, Dong-Woo

    2016-07-05

    Although various types of organs-on-chips have been introduced recently as tools for drug discovery, the current studies are limited in terms of fabrication methods. The fabrication methods currently available not only need a secondary cell-seeding process and result in severe protein absorption due to the material used, but also have difficulties in providing various cell types and extracellular matrix (ECM) environments for spatial heterogeneity in the organs-on-chips. Therefore, in this research, we introduce a novel 3D bioprinting method for organ-on-a-chip applications. With our novel 3D bioprinting method, it was possible to prepare an organ-on-a-chip in a simple one-step fabrication process. Furthermore, protein absorption on the printed platform was very low, which will lead to accurate measurement of metabolism and drug sensitivity. Moreover, heterotypic cell types and biomaterials were successfully used and positioned at the desired position for various organ-on-a-chip applications, which will promote full mimicry of the natural conditions of the organs. The liver organ was selected for the evaluation of the developed method, and liver function was shown to be significantly enhanced on the liver-on-a-chip, which was prepared by 3D bioprinting. Consequently, the results demonstrate that the suggested 3D bioprinting method is easier and more versatile for production of organs-on-chips.

  6. Dynamics of cells function on laser cell-chip system

    NASA Astrophysics Data System (ADS)

    Kushibiki, Toshihiro; Sano, Tomoko; Ishii, Katsunori; Yoshihashi-Suzuki, Sachiko; Awazu, Kunio

    2006-02-01

    A new type of cell-cultivation system based on laser processing has been developed for the on-chip cultivation of living cells. We introduce a "laser cell-chip", on which migration of cells, such as stem cells, tumor cells or immunocompetent cells, can be observed. A sheet prepared from epoxy resin was processed by KrF excimer laser (248 nm, 1.6 J/cm2) for preparation of microgrooved surfaces with various groove width, spacing, and depth. A laser cell-chip can make kinetic studies of cell migration depending on the concentration gradient of a chemoattractant. In this study, megakaryocytes were used for the migration on a groove of laser cell-chip by the concentration gradient of the stromal cell derived factor 1 (SDF-1/CXCL12). SDF-1/CXCL12 plays an important and unique role in the regulation of stem/progenitor cell trafficking. A megakaryocyte was migrated on a groove of laser cell-chip depending on the optical concentration gradient of SDF-1/CXCL12. Since SDF-1/CXCL12-induced migration of mature megakaryocyte was known to increase the platelet production in the bone marrow extravascular space, the diagnosis of cell migration on laser cell-chip could provide a new strategy to potentially reconstitute hematopoiesis and avoid life-threatening hemorrhage after myelosuppression or bone marrow failure.

  7. Controlled and tunable polymer particles' production using a single microfluidic device

    NASA Astrophysics Data System (ADS)

    Amoyav, Benzion; Benny, Ofra

    2018-04-01

    Microfluidics technology offers a new platform to control liquids under flow in small volumes. The advantage of using small-scale reactions for droplet generation along with the capacity to control the preparation parameters, making microfluidic chips an attractive technology for optimizing encapsulation formulations. However, one of the drawback in this methodology is the ability to obtain a wide range of droplet sizes, from sub-micron to microns using a single chip design. In fact, typically, droplet chips are used for micron-dimension particles, while nanoparticles' synthesis requires complex chips design (i.e., microreactors and staggered herringbone micromixer). Here, we introduce the development of a highly tunable and controlled encapsulation technique, using two polymer compositions, for generating particles ranging from microns to nano-size using the same simple single microfluidic chip design. Poly(lactic-co-glycolic acid) (PLGA 50:50) or PLGA/polyethylene glycol polymeric particles were prepared with focused-flow chip, yielding monodisperse particle batches. We show that by varying flow rate, solvent, surfactant and polymer composition, we were able to optimize particles' size and decrease polydispersity index, using simple chip designs with no further related adjustments or costs. Utilizing this platform, which offers tight tuning of particle properties, could offer an important tool for formulation development and can potentially pave the way towards a better precision nanomedicine.

  8. Infrared Spectroscopy for Rapid Characterization of Drill Core and Cutting Mineralogy

    NASA Astrophysics Data System (ADS)

    Calvin, W. M.; Kratt, C.; Kruse, F. A.

    2009-12-01

    Water geochemistry can vary with depth and location within a geothermal reservoir, owing to natural factors such as changing rock type, gas content, fluid source and temperature. The interaction of these variable fluids with the host rock will cause well known changes in alteration mineral assemblages that are commonly factored into the exploration of hydrothermal systems for economic metals, but are less utilized with regard to mapping borehole geology for geothermal energy production. Chemistry of geothermal fluids and rock alteration products can impact production factors such as pipeline corrosion and scaling and early studies explored the use of both silica and chlorites as geothermometers. Infrared spectroscopy is particularly good at identifying a wide variety of alteration minerals, especially in discrimination among clay minerals, with no sample preparation. The technique has been extensively used in the remote identification of materials, but is not commonly used on drill core or chips. We have performed several promising pilot studies that suggest the power of the technique to sample continuously and provide mineral logs akin to geophysical ones. We have surveyed a variety of samples, including drill chip boards, boxed core, and drill cuttings from envelopes, sample bottles and chip trays. This work has demonstrated that core and drill chips can be rapidly surveyed, acquiring spectra every few to tens of cm of section, or the vertical resolution of the chip tray (typically 10 feet). Depending on the sample type we can acquire spectral data over thousands of feet depth at high vertical resolution in a fraction of the time that is needed for traditional analytical methods such as XRD or TEM with better accuracy than traditional geologic drill or chip logging that uses visual inspection alone. We have successfully identified layered silicates such as illite, kaolinite, montmorillonite chlorite and prehnite, zeolites, opal, calcite, jarosite and iron oxides and hydroxides in geothermal drill samples. We are currently developing automated analysis techniques to convert this detailed spectral logging data into high-vertical-resolution mineral depth profiles that can be linked to lithology, stratigraphy, fracture zones and potential for geothermal production. Also in development are metrics that would link mapped mineralogy to known geothermometers such as Na-K, Mg depletion, discrimination among illite, montmorillonite, and beidellite, and kaolinite crystallinity. Identification of amorphous and crystalline silica components (chalcedony, crystobalite and quartz) can also constrain silica geothermometry. The degree of alteration and some mineral types have been shown to be a proxy for host rock permeability, natural circulation, and the potential for reservoir sealing. Analysis of alteration intensity is also under way. We will present a synthesis of results to date.

  9. High-throughput particle manipulation by hydrodynamic, electrokinetic, and dielectrophoretic effects in an integrated microfluidic chip

    PubMed Central

    Li, Shunbo; Li, Ming; Bougot-Robin, Kristelle; Cao, Wenbin; Yeung Yeung Chau, Irene; Li, Weihua; Wen, Weijia

    2013-01-01

    Integrating different steps on a chip for cell manipulations and sample preparation is of foremost importance to fully take advantage of microfluidic possibilities, and therefore make tests faster, cheaper and more accurate. We demonstrated particle manipulation in an integrated microfluidic device by applying hydrodynamic, electroosmotic (EO), electrophoretic (EP), and dielectrophoretic (DEP) forces. The process involves generation of fluid flow by pressure difference, particle trapping by DEP force, and particle redirect by EO and EP forces. Both DC and AC signals were applied, taking advantages of DC EP, EO and AC DEP for on-chip particle manipulation. Since different types of particles respond differently to these signals, variations of DC and AC signals are capable to handle complex and highly variable colloidal and biological samples. The proposed technique can operate in a high-throughput manner with thirteen independent channels in radial directions for enrichment and separation in microfluidic chip. We evaluated our approach by collecting Polystyrene particles, yeast cells, and E. coli bacteria, which respond differently to electric field gradient. Live and dead yeast cells were separated successfully, validating the capability of our device to separate highly similar cells. Our results showed that this technique could achieve fast pre-concentration of colloidal particles and cells and separation of cells depending on their vitality. Hydrodynamic, DC electrophoretic and DC electroosmotic forces were used together instead of syringe pump to achieve sufficient fluid flow and particle mobility for particle trapping and sorting. By eliminating bulky mechanical pumps, this new technique has wide applications for in situ detection and analysis. PMID:24404011

  10. High-throughput particle manipulation by hydrodynamic, electrokinetic, and dielectrophoretic effects in an integrated microfluidic chip.

    PubMed

    Li, Shunbo; Li, Ming; Bougot-Robin, Kristelle; Cao, Wenbin; Yeung Yeung Chau, Irene; Li, Weihua; Wen, Weijia

    2013-01-01

    Integrating different steps on a chip for cell manipulations and sample preparation is of foremost importance to fully take advantage of microfluidic possibilities, and therefore make tests faster, cheaper and more accurate. We demonstrated particle manipulation in an integrated microfluidic device by applying hydrodynamic, electroosmotic (EO), electrophoretic (EP), and dielectrophoretic (DEP) forces. The process involves generation of fluid flow by pressure difference, particle trapping by DEP force, and particle redirect by EO and EP forces. Both DC and AC signals were applied, taking advantages of DC EP, EO and AC DEP for on-chip particle manipulation. Since different types of particles respond differently to these signals, variations of DC and AC signals are capable to handle complex and highly variable colloidal and biological samples. The proposed technique can operate in a high-throughput manner with thirteen independent channels in radial directions for enrichment and separation in microfluidic chip. We evaluated our approach by collecting Polystyrene particles, yeast cells, and E. coli bacteria, which respond differently to electric field gradient. Live and dead yeast cells were separated successfully, validating the capability of our device to separate highly similar cells. Our results showed that this technique could achieve fast pre-concentration of colloidal particles and cells and separation of cells depending on their vitality. Hydrodynamic, DC electrophoretic and DC electroosmotic forces were used together instead of syringe pump to achieve sufficient fluid flow and particle mobility for particle trapping and sorting. By eliminating bulky mechanical pumps, this new technique has wide applications for in situ detection and analysis.

  11. Strength and Stiffness Development in Soft Soils: A FESEM aided Soil Microstructure Viewpoint

    NASA Astrophysics Data System (ADS)

    Wijeyesekera, D. C.; Ho, M. H.; Bai, X.; Bakar, I.

    2016-07-01

    This paper opens with an overview of the debatable definition of soft soil that goes beyond a (CH) organic / inorganic clay and OH peat to include weakly cemented periglacial deposits of loess and alike. It then outlines the findings obtained from stiffness test on cement-stabilised soft clay. The findings are complemented with a microstructure viewpoint obtained using field emission scanning electron microscope (FESEM). Research also comprised of making cylindrical stabilised clay samples, prepared in the laboratory with various rubber chips contents and cement, and then aged for 28 days. The samples were then subjected to unconfined compressive strength (UCS) test and observations were also made of its microstructure using the FESEM. The impact of the soil microstructure on the stiffness result was studied both with the stabilized soil and also of some of the natural undisturbed loess soils. Sustainability aspect and the potential of the use of rubber chips and sand as additives to cement stabilisation are also discussed. The overall test results indicated that rubber chips and sand contributed to the improvement in unconfined compressive strength (qu). The derogatory influence of moisture on the stiffness of the stabilised clay was studied simultaneously. SEM micrographs are presented that show bonding of cement, rubber chips/ sand and soft clay, granular units and aggregated / agglomerated units in loess. The paper concludes with observations on the dependence of soil microstructure on the soil strength and deformability and even collapsibility of the loess. Current practices adopted as engineering solutions to these challenging soils are outlined.

  12. A droplet-based microfluidic chip as a platform for leukemia cell lysate identification using surface-enhanced Raman scattering.

    PubMed

    Hassoun, Mohamed; Rüger, Jan; Kirchberger-Tolstik, Tatiana; Schie, Iwan W; Henkel, Thomas; Weber, Karina; Cialla-May, Dana; Krafft, Christoph; Popp, Jürgen

    2018-01-01

    A new approach is presented for cell lysate identification which uses SERS-active silver nanoparticles and a droplet-based microfluidic chip. Eighty-nanoliter droplets are generated by injecting silver nanoparticles, KCl as aggregation agent, and cell lysate containing cell constituents, such as nucleic acids, carbohydrates, metabolites, and proteins into a continuous flow of mineral oil. This platform enables accurate mixing of small volumes inside the meandering channels of the quartz chip and allows acquisition of thousands of SERS spectra with 785 nm excitation at an integration time of 1 s. Preparation of three batches of three leukemia cell lines demonstrated the experimental reproducibility. The main advantage of a high number of reproducible spectra is to apply statistics for large sample populations with robust classification results. A support vector machine with leave-one-batch-out cross-validation classified SERS spectra with sensitivities, specificities, and accuracies better than 99% to differentiate Jurkat, THP-1, and MONO-MAC-6 leukemia cell lysates. This approach is compared with previous published reports about Raman spectroscopy for leukemia detection, and an outlook is given for transfer to single cells. A quartz chip was designed for SERS at 785 nm excitation. Principal component analysis of SERS spectra clearly separates cell lysates using variations in band intensity ratios.

  13. Field-programmable lab-on-a-chip based on microelectrode dot array architecture.

    PubMed

    Wang, Gary; Teng, Daniel; Lai, Yi-Tse; Lu, Yi-Wen; Ho, Yingchieh; Lee, Chen-Yi

    2014-09-01

    The fundamentals of electrowetting-on-dielectric (EWOD) digital microfluidics are very strong: advantageous capability in the manipulation of fluids, small test volumes, precise dynamic control and detection, and microscale systems. These advantages are very important for future biochip developments, but the development of EWOD microfluidics has been hindered by the absence of: integrated detector technology, standard commercial components, on-chip sample preparation, standard manufacturing technology and end-to-end system integration. A field-programmable lab-on-a-chip (FPLOC) system based on microelectrode dot array (MEDA) architecture is presented in this research. The MEDA architecture proposes a standard EWOD microfluidic component called 'microelectrode cell', which can be dynamically configured into microfluidic components to perform microfluidic operations of the biochip. A proof-of-concept prototype FPLOC, containing a 30 × 30 MEDA, was developed by using generic integrated circuits computer aided design tools, and it was manufactured with standard low-voltage complementary metal-oxide-semiconductor technology, which allows smooth on-chip integration of microfluidics and microelectronics. By integrating 900 droplet detection circuits into microelectrode cells, the FPLOC has achieved large-scale integration of microfluidics and microelectronics. Compared to the full-custom and bottom-up design methods, the FPLOC provides hierarchical top-down design approach, field-programmability and dynamic manipulations of droplets for advanced microfluidic operations.

  14. The multifunctional application of microfluidic lab-on-a-chip surface enhanced Raman spectroscopy (LOC-SERS) within the field of bioanalytics

    NASA Astrophysics Data System (ADS)

    März, Anne; Mönch, Bettina; Walter, Angela; Bocklitz, Thomas; Schumacher, Wilm; Rösch, Petra; Kiehntopf, Michael; Popp, Jürgen

    2011-07-01

    This contribution will present a variety of applications of lab-on-a-chip surface enhanced Raman spectroscopy in the field of bioanalytic. Beside the quantification and online monitoring of drugs and pharmaceuticals, determination of enzyme activity and discrimination of bacteria are successfully carried out utilizing LOC-SERS. The online-monitoring of drugs using SERS in a microfluidic device is demonstrated for nicotine. The enzyme activity of thiopurine methyltransferase (TPMT) in lysed red blood cells is determined by SERS in a lab-on-a-chip device. To analyse the activity of TPMT the metabolism of 6-mercaptopurine to 6-methylmercaptopurine is investigated. The discrimination of bacteria on strain level is carried out with different E. coli strains. For the investigations, the bacteria are busted by ultra sonic to achieve a high information output. This sample preparation provides the possibility to detect SERS spectra containing information of the bacterial cell walls as well as of the cytoplasm. This contribution demonstrates the great potential of LOC-SERS in the field of bioanalytics.

  15. Fluorimetric urease inhibition assay on a multilayer microfluidic chip with immunoaffinity immobilized enzyme reactors.

    PubMed

    Zhang, Qin; Tang, Xiuwen; Hou, Fenghua; Yang, Jianping; Xie, Zhiyong; Cheng, Zhiyi

    2013-10-01

    We fabricated a three-layer polydimethylsiloxane (PDMS)-based microfluidic chip for realizing urease inhibition assay with sensitive fluorescence detection. Procedures such as sample prehandling, enzyme reaction, reagent mixing, fluorescence derivatization, and detection can be readily carried out. Urease reactors were prepared by adsorption of rabbit immunoglobulin G (IgG) and immunoreaction with urease-conjugated goat anti-rabbit IgG. Acetohydroxamic acid (AHA) as a competitive inhibitor of urease was tested on the chip. Microfluidically generated gradient concentrations of AHA with substrate (urea) were loaded into urease reactors. After incubation, the produced ammonia was transported out of reactors and then reacted with o-phthalaldehyde (OPA) to generate fluorescent products. Urease inhibition was indicated by a decrease in fluorescence signal detected by microplate reader. The IC50 value of AHA was determined and showed good agreement with that obtained in microplate. The presented device combines several steps of the analytical process with advantages of low reagent consumption, reduced analysis time, and ease of manipulation. This microfluidic approach can be extended to the screening of inhibitory compounds in drug discovery. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Integrated sample-to-detection chip for nucleic acid test assays.

    PubMed

    Prakash, R; Pabbaraju, K; Wong, S; Tellier, R; Kaler, K V I S

    2016-06-01

    Nucleic acid based diagnostic techniques are routinely used for the detection of infectious agents. Most of these assays rely on nucleic acid extraction platforms for the extraction and purification of nucleic acids and a separate real-time PCR platform for quantitative nucleic acid amplification tests (NATs). Several microfluidic lab on chip (LOC) technologies have been developed, where mechanical and chemical methods are used for the extraction and purification of nucleic acids. Microfluidic technologies have also been effectively utilized for chip based real-time PCR assays. However, there are few examples of microfluidic systems which have successfully integrated these two key processes. In this study, we have implemented an electro-actuation based LOC micro-device that leverages multi-frequency actuation of samples and reagents droplets for chip based nucleic acid extraction and real-time, reverse transcription (RT) PCR (qRT-PCR) amplification from clinical samples. Our prototype micro-device combines chemical lysis with electric field assisted isolation of nucleic acid in a four channel parallel processing scheme. Furthermore, a four channel parallel qRT-PCR amplification and detection assay is integrated to deliver the sample-to-detection NAT chip. The NAT chip combines dielectrophoresis and electrostatic/electrowetting actuation methods with resistive micro-heaters and temperature sensors to perform chip based integrated NATs. The two chip modules have been validated using different panels of clinical samples and their performance compared with standard platforms. This study has established that our integrated NAT chip system has a sensitivity and specificity comparable to that of the standard platforms while providing up to 10 fold reduction in sample/reagent volumes.

  17. Aflatoxin M1 in Tarhana chips.

    PubMed

    Özçam, Mustafa; Obuz, Ersel; Tosun, Halil

    2014-01-01

    Tarhana chips are a popular traditional fermented food consumed widely in the Kahramanmaraş region of Turkey. Tarhana chips are different from many other types of fermented food in that they are produced in the form of tortilla chips. Cereal and yoghurt are the main ingredients in Tarhana chips. Aflatoxin M1 (AFM1) levels in dairy and dairy-based products are of concern for human health. To investigate AFM1 contamination, a total of 40 samples were collected from Kahramanmaraş region and AFM1 levels were determined by competitive enzyme-linked immunosorbent assay (ELISA). Furthermore, physicochemical characteristics of Tarhana chips were investigated and compared with classic fried chips in terms of nutritional value. Based on data obtained from enzyme-linked immunosorbent assay, 21 (52.5%) out of 40 samples contained AFM1 in the range 0.5-36.6 ng/kg, so AFM1 levels of all samples were below the legal limit.

  18. Evaluation of the correctness of the feed selection based on the analysis of chip's shape

    NASA Astrophysics Data System (ADS)

    Chodor, Jaroslaw; Kukielka, Leon; Kaldunski, Pawel; Bohdal, Lukasz; Patyk, Radoslaw; Kulakowska, Agnieszka

    2018-05-01

    For the experiment needs, the own experiment plan was developed. The researches were carried out to determine the effect of variable, small feed on the chip's shape. To provide orthogonal free cutting special specimens were prepared. Obtained chips were divided according to shapes and also scanned using a scanning electron microscope (SEM). Conclusions from the experiments were given.

  19. Development of apple chips technology

    NASA Astrophysics Data System (ADS)

    Kowalska, Hanna; Marzec, Agata; Kowalska, Jolanta; Samborska, Kinga; Tywonek, Małgorzata; Lenart, Andrzej

    2018-05-01

    For develop of apple chips technology without chemical preservation osmotic dehydration in cherry or apple juice concentrates or fructooligosaccharide solutions and convection drying were used. Studies included the effect of dehydration on the mass transfer in apples and the quality of the final product. The temperature, type of osmotic solution and its concentration were changeable. The fruit were tested on mass transfer indicators, stability (water activity), texture (breaking test) and nutritional value (polyphenol content, acidity). Sensory evaluation was also performed. On this basis, the verification of all options was made and the most acceptable samples were selected. Concentration of osmotic solutions at 25°Brix limited solids gain in apples. Under these conditions, the phenomenon of osmosis caused 8-10 times greater water loss than solids gain. Increasing the concentration of solutions up to 50°Brix had a significantly greater impact on mass exchange in apples, compared to increasing the temperature from 40 to 60 °C. Osmotic dehydration before drying did not significantly affect the water activity but increase of the temperature negatively affected on breaking force of the chips. Chips obtained by osmotic dehydration of apples in a cherry concentrate solution contained significantly more polyphenols, and were characterized by a higher acidity than the variants obtained by dehydration in concentrated apple juice. Furthermore, they were marked by red color which has been thought as part of the attractiveness of the product. The least sensory acceptable chips were prepared using osmotic pre-treatment in cherry concentrated juice solution with the addition of fructooligosaccharide.

  20. Fully Integrated Microfluidic Device for Direct Sample-to-Answer Genetic Analysis

    NASA Astrophysics Data System (ADS)

    Liu, Robin H.; Grodzinski, Piotr

    Integration of microfluidics technology with DNA microarrays enables building complete sample-to-answer systems that are useful in many applications such as clinic diagnostics. In this chapter, a fully integrated microfluidic device [1] that consists of microfluidic mixers, valves, pumps, channels, chambers, heaters, and a DNA microarray sensor to perform DNA analysis of complex biological sample solutions is present. This device can perform on-chip sample preparation (including magnetic bead-based cell capture, cell preconcentration and purification, and cell lysis) of complex biological sample solutions (such as whole blood), polymerase chain reaction, DNA hybridization, and electrochemical detection. A few novel microfluidic techniques were developed and employed. A micromix-ing technique based on a cavitation microstreaming principle was implemented to enhance target cell capture from whole blood samples using immunomagnetic beads. This technique was also employed to accelerate DNA hybridization reaction. Thermally actuated paraffin-based microvalves were developed to regulate flows. Electrochemical pumps and thermopneumatic pumps were integrated on the chip to provide pumping of liquid solutions. The device is completely self-contained: no external pressure sources, fluid storage, mechanical pumps, or valves are necessary for fluid manipulation, thus eliminating possible sample contamination and simplifying device operation. Pathogenic bacteria detection from ~mL whole blood samples and single-nucleotide polymorphism analysis directly from diluted blood were demonstrated. The device provides a cost-effective solution to direct sample-to-answer genetic analysis, and thus has a potential impact in the fields of point-of-care genetic analysis, environmental testing, and biological warfare agent detection.

  1. From sample-to-answer: integrated genotyping and immunological analysis microfluidic platforms for the diagnostic and treatment of coeliac disease

    NASA Astrophysics Data System (ADS)

    Jung, M.; Höth, J.; Erwes, J.; Latta, D.; Strobach, X.; Hansen-Hagge, T.; Klemm, R.; Gärtner, C.; Demiris, T. M.; O'Sullivan, C.; Ritzi-Lehnert, M.; Drese, K. S.

    2011-02-01

    Taking advantage of microfluidics technology, a Lab-on-Chip system was developed offering the possibility of performing HLA (Human Leukocyte Antigen) typing to test genetic predisposition to coeliac disease and measure the level of immunodeficiency at the point-of-care. These analysis procedures are implemented on two different microfluidic cartridges, both having identical interfacial connections to the identical automated instrument. In order to assess the concentration of the targeted analytes in human blood, finger prick samples are processed to either extract genomic DNA carrying the coeliac disease gene or blood plasma containing the disease specific antibodies. We present here the different microfluidic modules integrated in a common platform, capable of automated sample preparation and analyte detection. In summary, this new microfluidic approach will dramatically reduce the costs of materials (polymer for the disposable chips and minute amount of bio-reagents) and minimize the time for analysis down to less than 20 minutes. In comparison to the state of the art detection of coeliac disease this work represents a tremendous improvement for the patient's quality of live and will significantly reduce the cost burden on the health care system.

  2. Recent progress in preparation and application of microfluidic chip electrophoresis

    NASA Astrophysics Data System (ADS)

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Yuan, Hua; Peng, Qiaohong; Tian, Chao

    2015-05-01

    Since its discovery in 1990, microfluidic chip electrophoresis (MCE) has allowed the development of applications with small size, fast analysis, low cost, high integration density and automatic level, which are easy to carry and have made commercialization efficient. MCE has been widely used in the areas of environmental protection, biochemistry, medicine and health, clinical testing, judicial expertise, food sanitation, pharmaceutical checking, drug testing, agrochemistry, biomedical engineering and life science. As one of the foremost fields in the research of capillary electrophoresis, MCE is the ultimate frontier to develop the miniaturized, integrated, automated all-in-one instruments needed in modern analytical chemistry. By adopting the advanced technologies of micro-machining, lasers and microelectronics, and the latest research achievements in analytical chemistry and biochemistry, the sampling, separation and detection systems of commonly used capillary electrophoresis are integrated with high densities onto glass, quartz, silicon or polymer wafers to form the MCE, which can finish the analysis of multi-step operations such as injection, enrichment, reaction, derivatization, separation, and collection of samples in a portable, efficient and super high speed manner. With reference to the different technological achievements in this area, the latest developments in MCE are reviewed in this article. The preparation mechanisms, surface modifications, and properties of different materials in MCE are compared, and the different sampling, separation and detection systems in MCE are summarized. The performance of MCE in analysis of fluorescent substance, metallic ion, sugar, medicine, nucleic acid, DNA, amino acid, polypeptide and protein is discussed, and the future direction of development is forecast.

  3. Lab-on-a-chip enabled HLA diagnostic: combined sample preparation and real time PCR for HLA-B57 diagnosis

    NASA Astrophysics Data System (ADS)

    Gärtner, Claudia; Becker, Holger; Hlawatsch, Nadine; Klemm, Richard; Moche, Christian; Schattschneider, Sebastian; Frank, Rainer; Willems, Andreas

    2015-05-01

    The diverse human HLA (human leukocyte antigen) system is responsible for antigen presentation and recognition. It is essential for the immune system to maintain a stable defense line, but also is also involved in autoimmunity as well as metabolic disease. HLA-haplotype (HLA-B27), for instance, is associated with inflammatory diseases such as Bechterew's disease. The administration of the HIV drug Abacavir in combination with another HLA-haplotype (HLAB57) is associated with severe hypersensitivity reactions. Accordingly, the HLA status has to be monitored for diagnosis or prior to start of therapy. Along this line, a miniaturized microfluidic platform has been developed allowing performing the complete analytical process from "sample-in" to "answer-out" in a point-of-care environment. The main steps of the analytical cascade inside the integrated system are blood cell lysis and DNA isolation, DNA purification, real-time PCR and quantitative monitoring of the rise of a fluorescent signal appearing during the PCR based sequence amplification. All bio-analytical steps were intended to be performed inside one chip and will be actuated, controlled and monitored by a matching device. This report will show that all required processes are established and tested and all device components work well and interact with the functional modules on the chips in a harmonized fashion.

  4. Trapping and Collection of Lymphocytes Using Microspot Array Chip and Magnetic Beads

    NASA Astrophysics Data System (ADS)

    Hashioka, Shingi; Obata, Tsutomu; Tokimitsu, Yoshiharu; Fujiki, Satoshi; Nakazato, Hiroyoshi; Muraguchi, Atsushi; Kishi, Hiroyuki; Tanino, Katsumi

    2006-04-01

    A microspot array chip, which has microspots of a magnetic thin film patterned on a glass substrate, was fabricated for trapping individual cells and for measuring their cellular response. The chip was easily fabricated by conventional semiconductor fabrication techniques on a mass production level as a disposable medical device. When a solution of lymphocyte-bound-magnetic beads was poured into the magnetized chip, each lymphocyte was trapped on each microspot of the magnetic thin film. The trapped cells were easily recovered from the chip using a micromanipulator. The micro-spot array chip can be utilized for arraying live cells and for measuring the response of each cell. The chip will be useful for preparing on array of different kinds of cells and for analyzing cellular response at the single cell level. The chip will be particularly useful for detecting antigen-specific B-lymphocytes and antigen-specific antibody complementary deoxyribonucleic acid (cDNA).

  5. Development of a package-free transparent disposable biosensor chip for simultaneous measurements of blood constituents and investigation of its storage stability.

    PubMed

    Nakamura, Hideaki; Tohyama, Kana; Tanaka, Masanori; Shinohara, Shouji; Tokunaga, Yuichi; Kurusu, Fumiyo; Koide, Satoshi; Gotoh, Masao; Karube, Isao

    2007-12-15

    A package-free transparent disposable biosensor chip was developed by a screen-printing technique. The biosensor chip was fabricated by stacking a substrate with two carbon electrodes on its surface, a spacer consisting of a resist layer and an adhesive layer, and a cover. The structure of the chip keeps the interior of the reaction-detecting section airtight until use. The chip is equipped with double electrochemical measuring elements for the simultaneous measurement of multiple items, and the reagent layer was developed in sample-feeding path. The sample-inlet port and air-discharge port are simultaneously opened by longitudinally folding in two biosensor units with a notch as a boundary. Then the shape of the chip is changed to a V-shape. The reaction-detecting section of the chip has a 1.0 microl sample volume for one biosensor unit. Excellent results were obtained with the chip in initial simultaneous chronoamperometric measurements of both glucose (r=1.00) and lactate (r=0.998) in the same samples. The stability of the enzyme sensor signals of the chip was estimated at ambient atmosphere on 8 testing days during a 6-month period. The results were compared with those obtained for an unpackaged chip used as a control. The package-free chip proved to be twice as good as the control chip in terms of the reproducibility of slopes from 16 calibration curves (one calibration curve: 0, 100, 300, 500 mg dl(-1) glucose; n=3) and 4.6 times better in terms of the reproducibility of correlation coefficients from the 16 calibration curves.

  6. Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation.

    PubMed

    Ashtiani, Dariush; Venugopal, Hari; Belousoff, Matthew; Spicer, Bradley; Mak, Johnson; Neild, Adrian; de Marco, Alex

    2018-04-06

    Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6-8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves. Copyright © 2018 Elsevier Inc. All rights reserved.

  7. The SmartBioPhone, a point of care vision under development through two European projects: OPTOLABCARD and LABONFOIL.

    PubMed

    Ruano-López, Jesus M; Agirregabiria, Maria; Olabarria, Garbiñe; Verdoy, Dolores; Bang, Dang D; Bu, Minqiang; Wolff, Anders; Voigt, Anja; Dziuban, Jan A; Walczak, Rafał; Berganzo, Javier

    2009-06-07

    This paper describes how sixteen partners from eight different countries across Europe are working together in two EU projects focused on the development of a point of care system. This system uses disposable Lab on a Chips (LOCs) that carry out the complete assay from sample preparation to result interpretation of raw samples. The LOC is either embedded in a flexible motherboard with the form of a smartcard (Labcard) or in a Skinpatch. The first project, OPTOLABCARD, extended and tested the use of a thick photoresit (SU-8) as a structural material to manufacture LOCs by lamination. This project produced several examples where SU-8 microfluidic circuitry revealed itself as a viable material for several applications, such as the integration on chip of a Polymerase Chain Reaction (PCR) that includes sample concentration, PCR amplification and optical detection of Salmonella spp. using clinical samples. The ongoing project, LABONFOIL, is using two results of OPTOLABCARD: the sample concentration method and the capability to fabricate flexible and ultra thin LOCs based on sheets instead of wafers. This rupture from the limited and expensive wafer surface heritage allows the development of a platform where LOCs are big enough to include all the sample preparation subcomponents at a low price. These LOCs will be used in four point of care applications: environment, food, cancer and drug monitoring. The user will obtain the results of the tests by connecting the Labcard/Skinpatch reader to a very popular interface (a smartphone), creating a new instrument namely "The SmartBioPhone". All standard smartphone capabilities will be at the disposal of the point of care instrument by a simple click. In order to guarantee the future mass production of these LOCs, the project will develop a large dry film equipment where LOCs will be fabricated at a low cost.

  8. Electrical field-induced extraction and separation techniques: promising trends in analytical chemistry--a review.

    PubMed

    Yamini, Yadollah; Seidi, Shahram; Rezazadeh, Maryam

    2014-03-03

    Sample preparation is an important issue in analytical chemistry, and is often a bottleneck in chemical analysis. So, the major incentive for the recent research has been to attain faster, simpler, less expensive, and more environmentally friendly sample preparation methods. The use of auxiliary energies, such as heat, ultrasound, and microwave, is one of the strategies that have been employed in sample preparation to reach the above purposes. Application of electrical driving force is the current state-of-the-art, which presents new possibilities for simplifying and shortening the sample preparation process as well as enhancing its selectivity. The electrical driving force has scarcely been utilized in comparison with other auxiliary energies. In this review, the different roles of electrical driving force (as a powerful auxiliary energy) in various extraction techniques, including liquid-, solid-, and membrane-based methods, have been taken into consideration. Also, the references have been made available, relevant to the developments in separation techniques and Lab-on-a-Chip (LOC) systems. All aspects of electrical driving force in extraction and separation methods are too specific to be treated in this contribution. However, the main aim of this review is to provide a brief knowledge about the different fields of analytical chemistry, with an emphasis on the latest efforts put into the electrically assisted membrane-based sample preparation systems. The advantages and disadvantages of these approaches as well as the new achievements in these areas have been discussed, which might be helpful for further progress in the future. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Comparison of the performance of Ion Torrent chips in noninvasive prenatal trisomy detection.

    PubMed

    Wang, Yanlin; Wen, Zujia; Shen, Jiawei; Cheng, Weiwei; Li, Jun; Qin, Xiaolan; Ma, Duan; Shi, Yongyong

    2014-07-01

    Semiconductor high-throughput sequencing, represented by Ion Torrent PGM/Proton, proves to be feasible in the noninvasive prenatal diagnosis of fetal aneuploidies. It is commendable that, with less data and relevant cost also, an accurate result can be achieved owing to the high sensitivity and specificity of such kind of technology. We conducted a comparative analysis of the performance of four different Ion chips in detecting fetal chromosomal aneuploidies. Eight maternal plasma DNA samples, including four pregnancies with normal fetuses and four with trisomy 21 fetuses, were sequenced on Ion Torrent 314/316/318/PI chips, respectively. Results such as read mapped ratio, correlation coefficient and phred quality score were calculated and parallelly compared. All samples were correctly classified even with low-throughput chip, and, among the four chips, the 316 chip had the highest read mapped ratio, correlation coefficient, mean read length and phred quality score. All chips were well consistent with each other. Our results showed that all Ion chips are applicable in noninvasive prenatal fetal aneuploidy diagnosis. We recommend researchers or clinicians to use the appropriate chip with barcoding technology on the basis of the sample number.

  10. Microfluidic filtration and extraction of pathogens from food samples by hydrodynamic focusing and inertial lateral migration.

    PubMed

    Clime, Liviu; Hoa, Xuyen D; Corneau, Nathalie; Morton, Keith J; Luebbert, Christian; Mounier, Maxence; Brassard, Daniel; Geissler, Matthias; Bidawid, Sabah; Farber, Jeff; Veres, Teodor

    2015-02-01

    Detecting pathogenic bacteria in food or other biological samples with lab-on-a-chip (LOC) devices requires several sample preparation steps prior to analysis which commonly involves cleaning complex sample matrices of large debris. This often underestimated step is important to prevent these larger particles from clogging devices and to preserve initial concentrations when LOC techniques are used to concentrate or isolate smaller target microorganisms for downstream analysis. In this context, we developed a novel microfluidic system for membrane-free cleaning of biological samples from debris particles by combining hydrodynamic focusing and inertial lateral migration effects. The microfluidic device is fabricated using thermoplastic elastomers being compatible with thermoforming fabrication techniques leading to low-cost single-use devices. Microfluidic chip design and pumping protocols are optimized by investigating diffusive losses numerically with coupled Navier-Stokes and convective-diffusion theoretical models. Stability of inertial lateral migration and separation of debris is assessed through fluorescence microscopy measurements with labelled particles serving as a model system. Efficiency of debris cleaning is experimentally investigated by monitoring microchip outlets with in situ optical turbidity sensors, while retention of targeted pathogens (i.e., Listeria monocytogenes) within the sample stream is assessed through bacterial culture techniques. Optimized pumping protocols can remove up to 50 % of debris from ground beef samples while percentage for preserved microorganisms can account for 95 % in relatively clean samples. However, comparison between inoculated turbid and clean samples (i.e., with and without ground beef debris) indicate some degree of interference between debris inertial lateral migration and hydrodynamic focusing of small microorganisms. Although this interference can lead to significant decrease in chip performance through loss of target bacteria, it remains possible to reach 70 % for sample recovery and more than 50 % for debris removal even in the most turbid samples tested. Due to the relatively simple design, the robustness of the inertial migration effect itself, the high operational flow rates and fabrication methods that leverage low-cost materials, the proposed device can have an impact on a wide range of applications where high-throughput separation of particles and biological species is of interest.

  11. Optical biosensor technologies for molecular diagnostics at the point-of-care

    NASA Astrophysics Data System (ADS)

    Schotter, Joerg; Schrittwieser, Stefan; Muellner, Paul; Melnik, Eva; Hainberger, Rainer; Koppitsch, Guenther; Schrank, Franz; Soulantika, Katerina; Lentijo-Mozo, Sergio; Pelaz, Beatriz; Parak, Wolfgang; Ludwig, Frank; Dieckhoff, Jan

    2015-05-01

    Label-free optical schemes for molecular biosensing hold a strong promise for point-of-care applications in medical research and diagnostics. Apart from diagnostic requirements in terms of sensitivity, specificity, and multiplexing capability, also other aspects such as ease of use and manufacturability have to be considered in order to pave the way to a practical implementation. We present integrated optical waveguide as well as magnetic nanoparticle based molecular biosensor concepts that address these aspects. The integrated optical waveguide devices are based on low-loss photonic wires made of silicon nitride deposited by a CMOS compatible plasma-enhanced chemical vapor deposition (PECVD) process that allows for backend integration of waveguides on optoelectronic CMOS chips. The molecular detection principle relies on evanescent wave sensing in the 0.85 μm wavelength regime by means of Mach-Zehnder interferometers, which enables on-chip integration of silicon photodiodes and, thus, the realization of system-on-chip solutions. Our nanoparticle-based approach is based on optical observation of the dynamic response of functionalized magneticcore/ noble-metal-shell nanorods (`nanoprobes') to an externally applied time-varying magnetic field. As target molecules specifically bind to the surface of the nanoprobes, the observed dynamics of the nanoprobes changes, and the concentration of target molecules in the sample solution can be quantified. This approach is suitable for dynamic real-time measurements and only requires minimal sample preparation, thus presenting a highly promising point-of-care diagnostic system. In this paper, we present a prototype of a diagnostic device suitable for highly automated sample analysis by our nanoparticle-based approach.

  12. Automated, Ultra-Sterile Solid Sample Handling and Analysis on a Chip

    NASA Technical Reports Server (NTRS)

    Mora, Maria F.; Stockton, Amanda M.; Willis, Peter A.

    2013-01-01

    There are no existing ultra-sterile lab-on-a-chip systems that can accept solid samples and perform complete chemical analyses without human intervention. The proposed solution is to demonstrate completely automated lab-on-a-chip manipulation of powdered solid samples, followed by on-chip liquid extraction and chemical analysis. This technology utilizes a newly invented glass micro-device for solid manipulation, which mates with existing lab-on-a-chip instrumentation. Devices are fabricated in a Class 10 cleanroom at the JPL MicroDevices Lab, and are plasma-cleaned before and after assembly. Solid samples enter the device through a drilled hole in the top. Existing micro-pumping technology is used to transfer milligrams of powdered sample into an extraction chamber where it is mixed with liquids to extract organic material. Subsequent chemical analysis is performed using portable microchip capillary electrophoresis systems (CE). These instruments have been used for ultra-highly sensitive (parts-per-trillion, pptr) analysis of organic compounds including amines, amino acids, aldehydes, ketones, carboxylic acids, and thiols. Fully autonomous amino acid analyses in liquids were demonstrated; however, to date there have been no reports of completely automated analysis of solid samples on chip. This approach utilizes an existing portable instrument that houses optics, high-voltage power supplies, and solenoids for fully autonomous microfluidic sample processing and CE analysis with laser-induced fluorescence (LIF) detection. Furthermore, the entire system can be sterilized and placed in a cleanroom environment for analyzing samples returned from extraterrestrial targets, if desired. This is an entirely new capability never demonstrated before. The ability to manipulate solid samples, coupled with lab-on-a-chip analysis technology, will enable ultraclean and ultrasensitive end-to-end analysis of samples that is orders of magnitude more sensitive than the ppb goal given in the Science Instruments.

  13. Portable and Reliable Surface-Enhanced Raman Scattering Silicon Chip for Signal-On Detection of Trace Trinitrotoluene Explosive in Real Systems.

    PubMed

    Chen, Na; Ding, Pan; Shi, Yu; Jin, Tengyu; Su, Yuanyuan; Wang, Houyu; He, Yao

    2017-05-02

    There is an increasing interest in the development of surface-enhanced Raman scattering (SERS) sensors for rapid and accurate on-site detection of hidden explosives. However, portable SERS methods for trace explosive detection in real systems remain scarce, mainly due to their relatively poor reliability and portability. Herein, we present the first demonstration of a portable silicon-based SERS analytical platform for signal-on detection of trace trinitrotoluene (TNT) explosives, which is made of silver nanoparticle (AgNP)-decorated silicon wafer chip (0.5 cm × 0.5 cm). In principle, under 514 nm excitation, the Raman signals of p-aminobenzenethiol (PABT) modified on the AgNP surface could be largely lit up due to the formation of electronic resonance-active TNT-PABT complex. In addition, the surface of AgNPs and silicon substrate-induced plasmon resonances also contribute the total SERS enhancement. For quantitative evaluation, the as-prepared chip features ultrahigh sensitivity [limit of detection is down to ∼1 pM (∼45.4 fg/cm 2 )] and adaptable reproducibility (relative standard deviation is less than 15%) in the detection of TNT standard solutions. More importantly, the developed chip can couple well with a hand-held Raman spectroscopic device using 785 nm excitation, suitable for qualitative analysis of trace TNT even at ∼10 -8 M level from environmental samples including lake water, soil, envelope, and liquor with a short data acquisition time (∼1 min). Furthermore, TNT vapors diffusing from TNT residues (∼10 -6 M) can be detected by using such a portable device, indicating its feasibility in determination of hidden samples.

  14. Microfluidic-Based sample chips for radioactive solutions

    DOE PAGES

    Tripp, J. L.; Law, J. D.; Smith, T. E.; ...

    2015-01-01

    Historical nuclear fuel cycle process sampling techniques required sample volumes ranging in the tens of milliliters. The radiation levels experienced by analytical personnel and equipment, in addition to the waste volumes generated from analysis of these samples, have been significant. These sample volumes also impacted accountability inventories of required analytes during process operations. To mitigate radiation dose and other issues associated with the historically larger sample volumes, a microcapillary sample chip was chosen for further investigation. The ability to obtain microliter volume samples coupled with a remote automated means of sample loading, tracking, and transporting to the analytical instrument wouldmore » greatly improve analytical efficiency while reducing both personnel exposure and radioactive waste volumes. Sample chip testing was completed to determine the accuracy, repeatability, and issues associated with the use of microfluidic sample chips used to supply µL sample volumes of lanthanide analytes dissolved in nitric acid for introduction to an analytical instrument for elemental analysis.« less

  15. Fabrication and characterization of SPR chips with the modified bovine serum albumin

    NASA Astrophysics Data System (ADS)

    Chen, Xing; Zhang, Lu-lu; Cui, Da-fu

    2016-03-01

    A facile surface plasmon resonance (SPR) chip is developed for small molecule determination and analysis. The SPR chip was prepared based on a self assembling principle, in which the modified bovine serum albumin (BSA) was directly self-assembled onto the bare gold surface. The surface morphology of the chip with the modified BSA was investigated by atomic force microscopy (AFM) and its optical properties were characterized. The surface binding capacity of the bare facile SPR chip with a uniform morphology is 8 times of that of the bare control SPR chip. Based on the experiments of immune reaction between cortisol antibody and cortisol derivative, the sensitivity of the facile SPR chip with the modified BSA is much higher than that of the control SPR chip with the un-modified BSA. The facile SPR chip has been successfully used to detect small molecules. The lowest detection limit is 5 ng/mL with a linear range of 5—100 ng/mL for cortisol analysis. The novel facile SPR chip can also be applied to detect other small molecules.

  16. Chip-scale white flip-chip light-emitting diode containing indium phosphide/zinc selenide quantum dots

    NASA Astrophysics Data System (ADS)

    Fan, Bingfeng; Yan, Linchao; Lao, Yuqin; Ma, Yanfei; Chen, Zimin; Ma, Xuejin; Zhuo, Yi; Pei, Yanli; Wang, Gang

    2017-08-01

    A method for preparing a quantum dot (QD)-white light-emitting diode (WLED) is reported. Holes were etched in the SiO2 layer deposited on the sapphire substrate of the flip-chip LED by inductively coupled plasma, and these holes were then filled with QDs. An ultraviolet-curable resin was then spin-coated on top of the QD-containing SiO2 layer, and the resin was cured to act as a protecting layer. The reflective sidewall structure minimized sidelight leakage. The fabrication of the QD-WLED is simple in preparation and compatible with traditional LED processes, which was the minimum size of the WLED chip-scale integrated package. InP/ZnS core-shell QDs were used as the converter in the WLED. A blue light-emitting diode with a flip-chip structure was used as the excitation source. The QD-WLED exhibited color temperatures from 5900 to 6400 K and Commission Internationale De L'Elcairage color coordinates from (0.315, 0.325) to (0.325, 0.317), under drive currents from 100 to 400 mA. The QD-WLED exhibited stable optoelectronic properties.

  17. A smartphone controlled handheld microfluidic liquid handling system.

    PubMed

    Li, Baichen; Li, Lin; Guan, Allan; Dong, Quan; Ruan, Kangcheng; Hu, Ronggui; Li, Zhenyu

    2014-10-21

    Microfluidics and lab-on-a-chip technologies have made it possible to manipulate small volume liquids with unprecedented resolution, automation and integration. However, most current microfluidic systems still rely on bulky off-chip infrastructures such as compressed pressure sources, syringe pumps and computers to achieve complex liquid manipulation functions. Here, we present a handheld automated microfluidic liquid handling system controlled by a smartphone, which is enabled by combining elastomeric on-chip valves and a compact pneumatic system. As a demonstration, we show that the system can automatically perform all the liquid handling steps of a bead-based HIV1 p24 sandwich immunoassay on a multi-layer PDMS chip without any human intervention. The footprint of the system is 6 × 10.5 × 16.5 cm, and the total weight is 829 g including battery. Powered by a 12.8 V 1500 mAh Li battery, the system consumed 2.2 W on average during the immunoassay and lasted for 8.7 h. This handheld microfluidic liquid handling platform is generally applicable to many biochemical and cell-based assays requiring complex liquid manipulation and sample preparation steps such as FISH, PCR, flow cytometry and nucleic acid sequencing. In particular, the integration of this technology with read-out biosensors may help enable the realization of the long-sought Tricorder-like handheld in vitro diagnostic (IVD) systems.

  18. Newborn screening by matrix-assisted laser desorption/ionization mass spectrometry based on parylene-matrix chip.

    PubMed

    Kim, Jo-Il; Noh, Joo-Yoon; Kim, Mira; Park, Jong-Min; Song, Hyun-Woo; Kang, Min-Jung; Pyun, Jae-Chul

    2017-08-01

    Newborn screening for diagnosis of phenylketonuria, homocystinuria, and maple syrup urine disease have been conducted by analyzing the concentration of target amino acids using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) based on parylene-matrix chip. Parylene-matrix chip was applied to MALDI-ToF MS analysis reducing the matrix peaks significantly at low mass-to-charge ratio range (m/z < 500). Reproducibility of inter-spot and intra-spot analyses of amino acids was less than 10%. Methanol extraction was adopted for simple and rapid sample preparation of serum before mass spectrometric analysis showing 13.3 to 45% of extraction efficiency. Calibration curves for diagnosis of neonatal metabolic disorders were obtained by analyzing methanol-extracted serum spiked with target amino acids using MALDI-ToF MS. They showed good linearity (R 2  > 0.98) and the LODs were ranging from 9.0 to 22.9 μg/mL. Effect of proteins in serum was estimated by comparing MALDI-ToF mass spectra of amino acids-spiked serum before and after the methanol extraction. Interference of other amino acids on analysis of target analyte was determined to be insignificant. From these results, MALDI-ToF MS based on parylene-matrix chip could be applicable to medical diagnosis of neonatal metabolic disorders. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Proteomic analysis of single mammalian cells enabled by microfluidic nanodroplet sample preparation and ultrasensitive nanoLC-MS.

    PubMed

    Zhu, Ying; Clair, Geremy; Chrisler, William; Shen, Yufeng; Zhao, Rui; Shukla, Anil; Moore, Ronald; Misra, Ravi; Pryhuber, Gloria; Smith, Richard; Ansong, Charles; Kelly, Ryan T

    2018-05-24

    We report on the quantitative proteomic analysis of single mammalian cells. Fluorescence-activated cell sorting was employed to deposit cells into a newly developed nanodroplet sample processing chip, after which samples were analysed by ultrasensitive nanoLC-MS. An average of ~670 protein groups were confidently identified from single HeLa cells, which is a far greater level of proteome coverage for single cells than has been previously reported. We demonstrate that the single cell proteomics platform can be used to differentiate cell types from enzyme-dissociated human lung primary cells and identify specific protein markers for epithelial and mesenchymal cells. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Asymmetric adiabatic couplers for fully-integrated broadband quantum-polarization state preparation.

    PubMed

    Chung, Hung-Pin; Huang, Kuang-Hsu; Wang, Kai; Yang, Sung-Lin; Yang, Shih-Yuan; Sung, Chun-I; Solntsev, Alexander S; Sukhorukov, Andrey A; Neshev, Dragomir N; Chen, Yen-Hung

    2017-12-04

    Spontaneous parametric down-conversion (SPDC) is a widely used method to generate entangled photons, enabling a range of applications from secure communication to tests of quantum physics. Integrating SPDC on a chip provides interferometric stability, allows to reduce a physical footprint, and opens a pathway to true scalability. However, dealing with different photon polarizations and wavelengths on a chip presents a number of challenging problems. In this work, we demonstrate an on-chip polarization beam-splitter based on z-cut titanium-diffused lithium niobate asymmetric adiabatic couplers (AAC) designed for integration with a type-II SPDC source. Our experimental measurements reveal unique polarization beam-splitting regime with the ability to tune the splitting ratios based on wavelength. In particular, we measured a splitting ratio of 17 dB over broadband regions (>60 nm) for both H- and V-polarized lights and a specific 50%/50% splitting ratio for a cross-polarized photon pair from the AAC. The results show that such a system can be used for preparing different quantum polarization-path states that are controllable by changing the phase-matching conditions in the SPDC over a broad band. Furthermore, we propose a fully integrated electro-optically tunable type-II SPDC polarization-path-entangled state preparation circuit on a single lithium niobate photonic chip.

  1. Rapid isolation of blood plasma using a cascaded inertial microfluidic device

    PubMed Central

    Robinson, M.; Hinsdale, T.; Coté, G.

    2017-01-01

    Blood, saliva, mucus, sweat, sputum, and other biological fluids are often hindered in their ability to be used in point-of-care (POC) diagnostics because their assays require some form of off-site sample pre-preparation to effectively separate biomarkers from larger components such as cells. The rapid isolation, identification, and quantification of proteins and other small molecules circulating in the blood plasma from larger interfering molecules are therefore particularly important factors for optical blood diagnostic tests, in particular, when using optical approaches that incur spectroscopic interference from hemoglobin-rich red blood cells (RBCs). In this work, a sequential spiral polydimethylsiloxane (PDMS) microfluidic device for rapid (∼1 min) on-chip blood cell separation is presented. The chip utilizes Dean-force induced migration via two 5-loop Archimedean spirals in series. The chip was characterized in its ability to filter solutions containing fluorescent beads and silver nanoparticles and further using blood solutions doped with a fluorescent protein. Through these experiments, both cellular and small molecule behaviors in the chip were assessed. The results exhibit an average RBC separation efficiency of ∼99% at a rate of 5.2 × 106 cells per second while retaining 95% of plasma components. This chip is uniquely suited for integration within a larger point-of-care diagnostic system for the testing of blood plasma, and the use of multiple filtering spirals allows for the tuning of filtering steps, making this device and the underlying technique applicable for a wide range of separation applications. PMID:28405258

  2. Wet-preserved hemp fibreboard properties improvement with veneering

    NASA Astrophysics Data System (ADS)

    Kirilovs, E.; Kukle, S.; Gusovius, H.-J.

    2015-03-01

    The initial research describes a new type of fiber boards for the furniture interior design, developed in cooperation with ATB (Leibniz-Institute for Agricultural Engineering) by using a new method of raw materials preparation and specific production technologies of ATB. The main raw materials are aerobically aged hemp stalks. The samples are made of hemp chips with a long preservation time and fastened together with the UF glue. Specimens are 8 mm thick and correspond to a medium-density fiberboard, fitting standard EN622. Due to the fact that non-veneered material can be used only in non-load-bearing constructions, material improving technologies were studied, such as increase of board density, increase of glue percentage, partially substitution of wet-preserved hemp chips with a dry hemp and/or wooden chips to equalize moisture content of obtained mixture. The particular article describes how the new material is veneered with the oak veneer obtaining three-ply composite board with the improved mechanical properties that allows to use these boards in a load-bearing constructions. Tests are performed with the veneered material to determine such parameters as static bending strength (MOR), modulus of elasticity in static bending (MOE), swelling in thickness and hardness.

  3. Deployable Laboratory Applications of Nano- and Bio-Technology (Applications de nanotechnologie et biotechnologie destinees a un laboratoire deployable)

    DTIC Science & Technology

    2014-10-01

    applications of present nano-/ bio -technology include advanced health and fitness monitoring, high-resolution imaging, new environmental sensor platforms...others areas where nano-/ bio -technology development is needed: • Sensors : Diagnostic and detection kits (gene-chips, protein-chips, lab-on-chips, etc...studies on chemo- bio nano- sensors , ultra-sensitive biochips (“lab-on-a-chip” and “cells-on-chips” devices) have been prepared for routine medical

  4. Facile synthesis of Prussian blue nanocubes/silver nanowires network as a water-based ink for the direct screen-printed flexible biosensor chips.

    PubMed

    Yang, Pengqi; Peng, Jingmeng; Chu, Zhenyu; Jiang, Danfeng; Jin, Wanqin

    2017-06-15

    The large-scale fabrication of nanocomposite based biosensors is always a challenge in the technology commercialization from laboratory to industry. In order to address this issue, we have designed a facile chemical method of fabricated nanocomposite ink applied to the screen-printed biosensor chip. This ink can be derived in the water through the in-situ growth of Prussian blue nanocubes (PBNCs) on the silver nanowires (AgNWs) to construct a composite nanostructure by a facile chemical method. Then a miniature flexible biosensor chip was screen-printed by using the prepared nanocomposite ink. Due to the synergic effects of the large specific surface area, high conductivity and electrocatalytic activity from AgNWs and PBNCs, the as-prepared biosensor chip exhibited a fast response (<3s), a wider linear response from 0.01 to 1.3mM with an ultralow LOD=5µm, and the ultrahigh sensitivities of 131.31 and 481.20µAmM -1 cm -2 for the detections of glucose and hydrogen peroxide (H 2 O 2 ), respectively. Furthermore, the biosensor chip exhibited excellent stability, good reproducibility and high anti-interference ability towards physiological substances under a very low working potential of -0.05. Hence, the proposed biosensor chip also showed a promising potential for the application in practical analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Double-cross hydrostatic pressure sample injection for chip CE: variable sample plug volume and minimum number of electrodes.

    PubMed

    Luo, Yong; Wu, Dapeng; Zeng, Shaojiang; Gai, Hongwei; Long, Zhicheng; Shen, Zheng; Dai, Zhongpeng; Qin, Jianhua; Lin, Bingcheng

    2006-09-01

    A novel sample injection method for chip CE was presented. This injection method uses hydrostatic pressure, generated by emptying the sample waste reservoir, for sample loading and electrokinetic force for dispensing. The injection was performed on a double-cross microchip. One cross, created by the sample and separation channels, is used for formation of a sample plug. Another cross, formed by the sample and controlling channels, is used for plug control. By varying the electric field in the controlling channel, the sample plug volume can be linearly adjusted. Hydrostatic pressure takes advantage of its ease of generation on a microfluidic chip, without any electrode or external pressure pump, thus allowing a sample injection with a minimum number of electrodes. The potential of this injection method was demonstrated by a four-separation-channel chip CE system. In this system, parallel sample separation can be achieved with only two electrodes, which is otherwise impossible with conventional injection methods. Hydrostatic pressure maintains the sample composition during the sample loading, allowing the injection to be free of injection bias.

  6. Purification of nanogram-range immunoprecipitated DNA in ChIP-seq application.

    PubMed

    Zhong, Jian; Ye, Zhenqing; Lenz, Samuel W; Clark, Chad R; Bharucha, Adil; Farrugia, Gianrico; Robertson, Keith D; Zhang, Zhiguo; Ordog, Tamas; Lee, Jeong-Heon

    2017-12-21

    Chromatin immunoprecipitation-sequencing (ChIP-seq) is a widely used epigenetic approach for investigating genome-wide protein-DNA interactions in cells and tissues. The approach has been relatively well established but several key steps still require further improvement. As a part of the procedure, immnoprecipitated DNA must undergo purification and library preparation for subsequent high-throughput sequencing. Current ChIP protocols typically yield nanogram quantities of immunoprecipitated DNA mainly depending on the target of interest and starting chromatin input amount. However, little information exists on the performance of reagents used for the purification of such minute amounts of immunoprecipitated DNA in ChIP elution buffer and their effects on ChIP-seq data. Here, we compared DNA recovery, library preparation efficiency, and ChIP-seq results obtained with several commercial DNA purification reagents applied to 1 ng ChIP DNA and also investigated the impact of conditions under which ChIP DNA is stored. We compared DNA recovery of ten commercial DNA purification reagents and phenol/chloroform extraction from 1 to 50 ng of immunopreciptated DNA in ChIP elution buffer. The recovery yield was significantly different with 1 ng of DNA while similar in higher DNA amounts. We also observed that the low nanogram range of purified DNA is prone to loss during storage depending on the type of polypropylene tube used. The immunoprecipitated DNA equivalent to 1 ng of purified DNA was subject to DNA purification and library preparation to evaluate the performance of four better performing purification reagents in ChIP-seq applications. Quantification of library DNAs indicated the selected purification kits have a negligible impact on the efficiency of library preparation. The resulting ChIP-seq data were comparable with the dataset generated by ENCODE consortium and were highly correlated between the data from different purification reagents. This study provides comparative data on commercial DNA purification reagents applied to nanogram-range immunopreciptated ChIP DNA and evidence for the importance of storage conditions of low nanogram-range purified DNA. We verified consistent high performance of a subset of the tested reagents. These results will facilitate the improvement of ChIP-seq methodology for low-input applications.

  7. Differentiation of Wines Treated with Wood Chips Based on Their Phenolic Content, Volatile Composition, and Sensory Parameters.

    PubMed

    Kyraleou, Maria; Kallithraka, Stamatina; Chira, Kleopatra; Tzanakouli, Eleni; Ligas, Ioannis; Kotseridis, Yorgos

    2015-12-01

    The effects of both wood chips addition and contact time on phenolic content, volatile composition, color parameters, and organoleptic character of red wine made by a native Greek variety (Agiorgitiko) were evaluated. For this purpose, chips from American, French, Slavonia oak, and Acacia were added in the wine after fermentation. A mixture consisting of 50% French and 50% Americal oak chips was also evaluated. In an attempt to categorize wine samples, various chemical parameters of wines and sensory parameters were studied after 1, 2, and 3 mo of contact time with chips. The results showed that regardless of the type of wood chips added in the wines, it was possible to differentiate the samples according to the contact time based on their phenolic composition and color parameters. In addition, wood-extracted volatile compounds seem to be the critical parameter that could separate the samples according to the wood type. The wines that were in contact with Acacia and Slavonia chips could be separated from the rest mainly due to their distinct sensory characters. © 2015 Institute of Food Technologists®

  8. Analysis of peptides using an integrated microchip HPLC-MS/MS system.

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kirby, Brian J.; Chirica, Gabriela S.; Reichmuth, David S.

    Hyphendated LC-MS techniques are quickly becoming the standard tool for protemic analyses. For large homogeneous samples, bulk processing methods and capillary injection and separation techniques are suitable. However, for analysis of small or heterogeneous samples, techniques that can manipulate picoliter samples without dilution are required or samples will be lost or corrupted; further, static nanospray-type flowrates are required to maximize SNR. Microchip-level integration of sample injection with separation and mass spectrometry allow small-volume analytes to be processed on chip and immediately injected without dilution for analysis. An on-chip HPLC was fabricated using in situ polymerization of both fixed and mobilemore » polymer monoliths. Integration of the chip with a nanospray MS emitter enables identification of peptides by the use of tandem MS. The chip is capable of analyzing of very small sample volumes (< 200 pl) in short times (< 3 min).« less

  9. Spectroscopic analysis of hot-water- and dilute-acid-extracted hardwood and softwood chips

    NASA Astrophysics Data System (ADS)

    Lehto, Joni; Louhelainen, Jarmo; Huttunen, Marko; Alén, Raimo

    2017-09-01

    Hot-water and dilute sulfuric acid pretreatments were performed prior to chemical pulping for silver/white birch (Betula pendula/B. pubescens) and Scots pine (Pinus sylvestris) chips to determine if varying pretreatment conditions on the original wood material were detectable via attenuated total reflectance (ATR) infrared spectroscopy. Pretreatment conditions varied with respect to temperature (130 °C and 150 °C) and treatment time (from 30 min to 120 min). The effects of the pretreatments on the composition of wood chips were determined by ATR infrared spectroscopy. The spectral data were compared to those determined by common wood chemistry analyses to evaluate the suitability of ATR spectroscopy method for rapid detection of changes in the wood chemical composition caused by different pretreatment conditions. In addition to determining wood species-dependent differences in the wood chemical composition, analytical results indicated that most essential lignin- and carbohydrates-related phenomena taking place during hot-water and acidic pretreatments could be described by applying this simple spectral method requiring only a small sample amount and sample preparation. Such information included, for example, the cleavage of essential lignin bonds (i.e., mainly β-O-4 linkages in guaiacyl and syringyl lignin) and formation of newly condensed lignin structures under different pretreatment conditions. Carbohydrate analyses indicated significant removal of hemicelluloses (especially hardwood xylan) and hemicelluloses-derived acetyl groups during the pretreatments, but they also confirmed the highly resistant nature of cellulose towards mild pretreatments.

  10. Spectroscopic analysis of hot-water- and dilute-acid-extracted hardwood and softwood chips.

    PubMed

    Lehto, Joni; Louhelainen, Jarmo; Huttunen, Marko; Alén, Raimo

    2017-09-05

    Hot-water and dilute sulfuric acid pretreatments were performed prior to chemical pulping for silver/white birch (Betula pendula/B. pubescens) and Scots pine (Pinus sylvestris) chips to determine if varying pretreatment conditions on the original wood material were detectable via attenuated total reflectance (ATR) infrared spectroscopy. Pretreatment conditions varied with respect to temperature (130°C and 150°C) and treatment time (from 30min to 120min). The effects of the pretreatments on the composition of wood chips were determined by ATR infrared spectroscopy. The spectral data were compared to those determined by common wood chemistry analyses to evaluate the suitability of ATR spectroscopy method for rapid detection of changes in the wood chemical composition caused by different pretreatment conditions. In addition to determining wood species-dependent differences in the wood chemical composition, analytical results indicated that most essential lignin- and carbohydrates-related phenomena taking place during hot-water and acidic pretreatments could be described by applying this simple spectral method requiring only a small sample amount and sample preparation. Such information included, for example, the cleavage of essential lignin bonds (i.e., mainly β-O-4 linkages in guaiacyl and syringyl lignin) and formation of newly condensed lignin structures under different pretreatment conditions. Carbohydrate analyses indicated significant removal of hemicelluloses (especially hardwood xylan) and hemicelluloses-derived acetyl groups during the pretreatments, but they also confirmed the highly resistant nature of cellulose towards mild pretreatments. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Development of a versatile lab-on-a-chip enzyme assay platform for pathogen detection in CBRNE scenarios

    NASA Astrophysics Data System (ADS)

    Klemm, Richard; Schattschneider, Sebastian; Jahn, Tobias; Hlawatsch, Nadine; Julich, Sandra; Becker, Holger; Gärtner, Claudia

    2013-05-01

    The ability to integrate complete assays on a microfluidic chip helps to greatly simplify instrument requirements and allows the use of lab-on-a-chip technology in the field. A core application for such field-portable systems is the detection of pathogens in a CBRNE scenario such as permanent monitoring of airborne pathogens, e.g. in metro stations or hospitals etc. As one assay methodology for the pathogen identification, enzymatic assays were chosen. In order evaluate different detection strategies, the realized on-chip enzyme assay module has been designed as a general platform chip. In all application cases, the assays are based on immobilized probes located in microfluidic channels. Therefore a microfluidic chip was realized containing a set of three individually addressable channels, not only for detection of the sample itself also to have a set of references for a quantitative analysis. It furthermore includes two turning valves and a waste container for clear and sealed storage of potential pathogenic liquids to avoid contamination of the environment. All liquids remain in the chip and can be disposed of in proper way subsequently to the analysis. The chip design includes four inlet ports consisting of one sample port (Luer interface) and three mini Luer interfaces for fluidic support of e.g. washing buffer, substrate and enzyme solution. The sample can be applied via a special, sealable sampling vessel with integrated female Luer interface. Thereby also pre-anaytical contamination of the environment can be provided. Other reagents that are required for analysis will be stored off chip.

  12. Optics-Integrated Microfluidic Platforms for Biomolecular Analyses

    PubMed Central

    Bates, Kathleen E.; Lu, Hang

    2016-01-01

    Compared with conventional optical methods, optics implemented on microfluidic chips provide small, and often much cheaper ways to interrogate biological systems from the level of single molecules up to small model organisms. The optical probing of single molecules has been used to investigate the mechanical properties of individual biological molecules; however, multiplexing of these measurements through microfluidics and nanofluidics confers many analytical advantages. Optics-integrated microfluidic systems can significantly simplify sample processing and allow a more user-friendly experience; alignments of on-chip optical components are predetermined during fabrication and many purely optical techniques are passively controlled. Furthermore, sample loss from complicated preparation and fluid transfer steps can be virtually eliminated, a particularly important attribute for biological molecules at very low concentrations. Excellent fluid handling and high surface area/volume ratios also contribute to faster detection times for low abundance molecules in small sample volumes. Although integration of optical systems with classical microfluidic analysis techniques has been limited, microfluidics offers a ready platform for interrogation of biophysical properties. By exploiting the ease with which fluids and particles can be precisely and dynamically controlled in microfluidic devices, optical sensors capable of unique imaging modes, single molecule manipulation, and detection of minute changes in concentration of an analyte are possible. PMID:27119629

  13. Bubble-free on-chip continuous-flow polymerase chain reaction: concept and application.

    PubMed

    Wu, Wenming; Kang, Kyung-Tae; Lee, Nae Yoon

    2011-06-07

    Bubble formation inside a microscale channel is a significant problem in general microfluidic experiments. The problem becomes especially crucial when performing a polymerase chain reaction (PCR) on a chip which is subject to repetitive temperature changes. In this paper, we propose a bubble-free sample injection scheme applicable for continuous-flow PCR inside a glass/PDMS hybrid microfluidic chip, and attempt to provide a theoretical basis concerning bubble formation and elimination. Highly viscous paraffin oil plugs are employed in both the anterior and posterior ends of a sample plug, completely encapsulating the sample and eliminating possible nucleation sites for bubbles. In this way, internal channel pressure is increased, and vaporization of the sample is prevented, suppressing bubble formation. Use of an oil plug in the posterior end of the sample plug aids in maintaining a stable flow of a sample at a constant rate inside a heated microchannel throughout the entire reaction, as compared to using an air plug. By adopting the proposed sample injection scheme, we demonstrate various practical applications. On-chip continuous-flow PCR is performed employing genomic DNA extracted from a clinical single hair root sample, and its D1S80 locus is successfully amplified. Also, chip reusability is assessed using a plasmid vector. A single chip is used up to 10 times repeatedly without being destroyed, maintaining almost equal intensities of the resulting amplicons after each run, ensuring the reliability and reproducibility of the proposed sample injection scheme. In addition, the use of a commercially-available and highly cost-effective hot plate as a potential candidate for the heating source is investigated.

  14. Performance Evaluation of Protein Chip Assay for Rapid Diagnosis of Hepatitis C Virus Infection in Injection Drug Abusers.

    PubMed

    Dai, Zhiyan; Shu, Xin; Li, Gang; Xi, Weidong

    2018-04-01

    We evaluated the performance of a protein chip assay for the detection of antibodies to hepatitis C virus (HCV) peptides among injection drug abusers (IDAs) by comparing the assay with existing methods, including enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR), and recombinant immunoblot assay (RIBA). Seventy serum samples collected from IDAs were analyzed by protein chip assay. ELISA, RT-PCR, and RIBA assay were used to validate the results. The protein chips could detect different peptides' antibodies against HCV-C, HCV-NS3, HCV-NS4, HCV-NS5, and HCV-mix antigen; no cross reactivity between antigens was observed. Results of the protein-chip assay were compared with those of ELISA. Any inconsistency in results was validated by both RT-PCR and RIBA. Concordance between the results of the protein-chip assay and ELISA was 96.3% for positive samples and 100% for negative samples. The protein chip had a higher specificity than ELISA, a higher sensitivity than RTPCR, and similar specificity and sensitivity as compared to RIBA. The limit of detection of HCV antibodies in the protein chip assay was examined and calculated by incubation of model array with different dilutions. The protein chip assay required smaller amounts of both samples and reagents; it detected serum antibody in sample quantity as low as 3 ng/mL and at antibody dilution as low as 1:1000; its cost was low as well. The positive rates in the antiC, anti-NS3, anti-NS4, and anti-NS5 groups were significantly associated with levels of HCV RNA and the viral load. The HCV RNA and protein chip positive rate in the injection equipment-sharing group was higher than that in the non-injection equipment-sharing group. The protein chip assay is a faster and simpler approach to simultaneously screen for all HCV peptide antibodies accurately and to provide a rapid diagnosis of HCV infection in IDAs. The dominant positive HCV peptide antibodies were significantly associated with HCV RNA load, especially in the injection equipment-sharing group.

  15. Stratified randomization controls better for batch effects in 450K methylation analysis: a cautionary tale.

    PubMed

    Buhule, Olive D; Minster, Ryan L; Hawley, Nicola L; Medvedovic, Mario; Sun, Guangyun; Viali, Satupaitea; Deka, Ranjan; McGarvey, Stephen T; Weeks, Daniel E

    2014-01-01

    Batch effects in DNA methylation microarray experiments can lead to spurious results if not properly handled during the plating of samples. Two pilot studies examining the association of DNA methylation patterns across the genome with obesity in Samoan men were investigated for chip- and row-specific batch effects. For each study, the DNA of 46 obese men and 46 lean men were assayed using Illumina's Infinium HumanMethylation450 BeadChip. In the first study (Sample One), samples from obese and lean subjects were examined on separate chips. In the second study (Sample Two), the samples were balanced on the chips by lean/obese status, age group, and census region. We used methylumi, watermelon, and limma R packages, as well as ComBat, to analyze the data. Principal component analysis and linear regression were, respectively, employed to identify the top principal components and to test for their association with the batches and lean/obese status. To identify differentially methylated positions (DMPs) between obese and lean males at each locus, we used a moderated t-test. Chip effects were effectively removed from Sample Two but not Sample One. In addition, dramatic differences were observed between the two sets of DMP results. After "removing" batch effects with ComBat, Sample One had 94,191 probes differentially methylated at a q-value threshold of 0.05 while Sample Two had zero differentially methylated probes. The disparate results from Sample One and Sample Two likely arise due to the confounding of lean/obese status with chip and row batch effects. Even the best possible statistical adjustments for batch effects may not completely remove them. Proper study design is vital for guarding against spurious findings due to such effects.

  16. Stratified randomization controls better for batch effects in 450K methylation analysis: a cautionary tale

    PubMed Central

    Buhule, Olive D.; Minster, Ryan L.; Hawley, Nicola L.; Medvedovic, Mario; Sun, Guangyun; Viali, Satupaitea; Deka, Ranjan; McGarvey, Stephen T.; Weeks, Daniel E.

    2014-01-01

    Background: Batch effects in DNA methylation microarray experiments can lead to spurious results if not properly handled during the plating of samples. Methods: Two pilot studies examining the association of DNA methylation patterns across the genome with obesity in Samoan men were investigated for chip- and row-specific batch effects. For each study, the DNA of 46 obese men and 46 lean men were assayed using Illumina's Infinium HumanMethylation450 BeadChip. In the first study (Sample One), samples from obese and lean subjects were examined on separate chips. In the second study (Sample Two), the samples were balanced on the chips by lean/obese status, age group, and census region. We used methylumi, watermelon, and limma R packages, as well as ComBat, to analyze the data. Principal component analysis and linear regression were, respectively, employed to identify the top principal components and to test for their association with the batches and lean/obese status. To identify differentially methylated positions (DMPs) between obese and lean males at each locus, we used a moderated t-test. Results: Chip effects were effectively removed from Sample Two but not Sample One. In addition, dramatic differences were observed between the two sets of DMP results. After “removing” batch effects with ComBat, Sample One had 94,191 probes differentially methylated at a q-value threshold of 0.05 while Sample Two had zero differentially methylated probes. The disparate results from Sample One and Sample Two likely arise due to the confounding of lean/obese status with chip and row batch effects. Conclusion: Even the best possible statistical adjustments for batch effects may not completely remove them. Proper study design is vital for guarding against spurious findings due to such effects. PMID:25352862

  17. Spike-In Normalization of ChIP Data Using DNA-DIG-Antibody Complex.

    PubMed

    Eberle, Andrea B

    2018-01-01

    Chromatin immunoprecipitation (ChIP) is a widely used method to determine the occupancy of specific proteins within the genome, helping to unravel the function and activity of specific genomic regions. In ChIP experiments, normalization of the obtained data by a suitable internal reference is crucial. However, particularly when comparing differently treated samples, such a reference is difficult to identify. Here, a simple method to improve the accuracy and reliability of ChIP experiments by the help of an external reference is described. An artificial molecule, composed of a well-defined digoxigenin (DIG) labeled DNA fragment in complex with an anti-DIG antibody, is synthesized and added to each chromatin sample before immunoprecipitation. During the ChIP procedure, the DNA-DIG-antibody complex undergoes the same treatments as the chromatin and is therefore purified and quantified together with the chromatin of interest. This external reference compensates for variability during the ChIP routine and improves the similarity between replicates, thereby emphasizing the biological differences between samples.

  18. Tough silk fibers prepared in air using a biomimetic microfluidic chip.

    PubMed

    Luo, Jie; Zhang, Lele; Peng, Qingfa; Sun, Mengjie; Zhang, Yaopeng; Shao, Huili; Hu, Xuechao

    2014-05-01

    Microfluidic chips with single channel were built to mimic the shear and elongation conditions in the spinning apparatus of spider and silkworm. Silk fibers dry-spun from regenerated silk fibroin (RSF) aqueous solution using the chip could be tougher than degummed natural silk. The artificial silk exhibited a breaking strength up to 614 MPa, a breaking elongation up to 27% and a breaking energy of 101 kJ/kg. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. The dosimetric properties of phosphate glass systems prepared by different chemical nanomaterials.

    PubMed

    Abdelhalim, Mohamed Anwar K; Al-Shamrani, Bandar Mora

    2016-12-01

    The synthesis and characterization of glass systems were carried out using prepared nanocrystals injected into a glass matrix as a thermoluminescence (TL) activator using the melt-quenching method. Sample 1 was prepared as [40P 2 O 5 50BaO:2.5MgO, 2.5Na 2 O, 5TiO 2 ], sample 2 as [37.5P 2 O 5 37.5CaO:25TiO 2 ] and sample 3 as [50P 2 O 5 -50Li 2 O]. Formation of the synthesized compound was confirmed by studying the X-ray diffraction (XRD) patterns and scanning electron microscopy (SEM) images. An annealing procedure was carried out for 1 h at 400 °C. The glow curve position and shape shifted dramatically and linearly to the higher temperature values on increasing the heating rate. A heating rate of 30 °C/s was the most suitable for obtaining a high TL response. Samples 2 and 3 have the highest TL response, which approached the effective atomic number (Z eff ) of natural bone. The observed TL sensitivity of the prepared samples 2 and 3 is less than that of commercially available 'TLD-200 chips' and LiF:Mg,Ti (TLD-100) phosphor. Sample [37.5P 2 O 5 37.5CaO:25TiO 2 ] would be useful in personal and environmental dosimetry for measuring high doses of gamma radiation. Sample [50P 2 O 5 -50Li 2 O] is a good dosimeter, although it requires the addition of an appropriate transitional metal (activator) to overcome the problem of high fading. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  20. Changes in the Chemical Composition of Plum Distillate During Maturation with Oak Chips under Different Conditions

    PubMed Central

    2017-01-01

    Summary This study investigates the effect of ageing on the qualitative and quantitative composition of plum distillate in contact with oak wood chips. Maturation was performed with lightly toasted French oak (Quercus sessiflora and Quercus robur) chips or oak chips made from fragments of empty barrels that had been used for ageing cognac. The effects of oak chip dose, process temperature, ageing system (static or circulatory) and ultrasound treatment were assessed. Maturation of plum distillate samples with oak chips resulted in higher levels of extractable organics (including tannins) and colour changes, which were correlated with the type and dose of oak chips, and the conditions of maturation. The content of sugars such as glucose, xylose and arabinose also increased, depending on the conditions and type of oak chips. Degradation of lignin resulted in liberation of sinapaldehyde, syringaldehyde, coniferaldehyde and vanillin, with intensities depending on the applied parameters. In terms of volatiles, decreases in the concentration of higher alcohols and aliphatic aldehydes were observed in the majority of maturation experiments, while concentrations of furanic aldehydes increased depending on the type and dose of oak chips, as well as on the conditions of maturation. The quantities of esters such as ethyl acetate decreased in the majority of experimental variants, whereas concentrations of ethyl caproate, ethyl caprylate and ethyl caprate increased gradually. Some phenols and lactones were detected in all matured samples, with the lowest levels found in the samples aged with oak chips made from cognac barrels. PMID:29089848

  1. Changes in the Chemical Composition of Plum Distillate During Maturation with Oak Chips under Different Conditions.

    PubMed

    Balcerek, Maria; Pielech-Przybylska, Katarzyna; Dziekońska-Kubczak, Urszula; Patelski, Piotr; Strąk, Ewelina

    2017-09-01

    This study investigates the effect of ageing on the qualitative and quantitative composition of plum distillate in contact with oak wood chips. Maturation was performed with lightly toasted French oak ( Quercus sessiflora and Quercus robur ) chips or oak chips made from fragments of empty barrels that had been used for ageing cognac. The effects of oak chip dose, process temperature, ageing system (static or circulatory) and ultrasound treatment were assessed. Maturation of plum distillate samples with oak chips resulted in higher levels of extractable organics (including tannins) and colour changes, which were correlated with the type and dose of oak chips, and the conditions of maturation. The content of sugars such as glucose, xylose and arabinose also increased, depending on the conditions and type of oak chips. Degradation of lignin resulted in liberation of sinapaldehyde, syringaldehyde, coniferaldehyde and vanillin, with intensities depending on the applied parameters. In terms of volatiles, decreases in the concentration of higher alcohols and aliphatic aldehydes were observed in the majority of maturation experiments, while concentrations of furanic aldehydes increased depending on the type and dose of oak chips, as well as on the conditions of maturation. The quantities of esters such as ethyl acetate decreased in the majority of experimental variants, whereas concentrations of ethyl caproate, ethyl caprylate and ethyl caprate increased gradually. Some phenols and lactones were detected in all matured samples, with the lowest levels found in the samples aged with oak chips made from cognac barrels.

  2. The characterization of four gene expression analysis in circulating tumor cells made by Multiplex-PCR from the AdnaTest kit on the lab-on-a-chip Agilent DNA 1000 platform.

    PubMed

    Škereňová, Markéta; Mikulová, Veronika; Čapoun, Otakar; Zima, Tomáš

    2016-01-01

    Nowadays, on-a-chip capillary electrophoresis is a routine method for the detection of PCR fragments. The Agilent 2100 Bioanalyzer was one of the first commercial devices in this field. Our project was designed to study the characteristics of Agilent DNA 1000 kit in PCR fragment analysis as a part of circulating tumour cell (CTC) detection technique. Despite the common use of this kit a complex analysis of the results from a long-term project is still missing. A commercially available Agilent DNA 1000 kit was used as a final step in the CTC detection (AdnaTest) for the determination of the presence of PCR fragments generated by Multiplex PCR. Data from 30 prostate cancer patients obtained during two years of research were analyzed to determine the trueness and precision of the PCR fragment size determination. Additional experiments were performed to demonstrate the precision (repeatability, reproducibility) and robustness of PCR fragment concentration determination. The trueness and precision of the size determination was below 3% and 2% respectively. The repeatability of the concentration determination was below 15%. The difference in concentration determination increases when Multiplex-PCR/storage step is added between the two measurements of one sample. The characteristics established in our study are in concordance with the manufacturer's specifications established for a ladder as a sample. However, the concentration determination may vary depending on chip preparation, sample storage and concentration. The 15% variation of concentration determination repeatability was shown to be partly proportional and can be suppressed by proper normalization.

  3. A mechanical cell disruption microfluidic platform based on an on-chip micropump.

    PubMed

    Cheng, Yinuo; Wang, Yue; Wang, Zhiyuan; Huang, Liang; Bi, Mingzhao; Xu, Wenxiao; Wang, Wenhui; Ye, Xiongying

    2017-03-01

    Cell disruption plays a vital role in detection of intracellular components which contain information about genetic and disease characteristics. In this paper, we demonstrate a novel microfluidic platform based on an on-chip micropump for mechanical cell disruption and sample transport. A 50  μ l cell sample can be effectively lysed through on-chip multi-disruption in 36 s without introducing any chemical agent and suffering from clogging by cellular debris. After 30 cycles of circulating disruption, 80.6% and 90.5% cell disruption rates were achieved for the HEK293 cell sample and human natural killer cell sample, respectively. Profiting from the feature of pump-on-chip, the highly integrated platform enables more convenient and cost-effective cell disruption for the analysis of intracellular components.

  4. A mechanical cell disruption microfluidic platform based on an on-chip micropump

    PubMed Central

    Cheng, Yinuo; Wang, Yue; Wang, Zhiyuan; Bi, Mingzhao; Xu, Wenxiao; Ye, Xiongying

    2017-01-01

    Cell disruption plays a vital role in detection of intracellular components which contain information about genetic and disease characteristics. In this paper, we demonstrate a novel microfluidic platform based on an on-chip micropump for mechanical cell disruption and sample transport. A 50 μl cell sample can be effectively lysed through on-chip multi-disruption in 36 s without introducing any chemical agent and suffering from clogging by cellular debris. After 30 cycles of circulating disruption, 80.6% and 90.5% cell disruption rates were achieved for the HEK293 cell sample and human natural killer cell sample, respectively. Profiting from the feature of pump-on-chip, the highly integrated platform enables more convenient and cost-effective cell disruption for the analysis of intracellular components. PMID:28798848

  5. Capillary-driven surface-enhanced Raman scattering (SERS)-based microfluidic chip for abrin detection

    NASA Astrophysics Data System (ADS)

    Yang, Hao; Deng, Min; Ga, Shan; Chen, Shouhui; Kang, Lin; Wang, Junhong; Xin, Wenwen; Zhang, Tao; You, Zherong; An, Yuan; Wang, Jinglin; Cui, Daxiang

    2014-03-01

    Herein, we firstly demonstrate the design and the proof-of-concept use of a capillary-driven surface-enhanced Raman scattering (SERS)-based microfluidic chip for abrin detection. The micropillar array substrate was etched and coated with a gold film by microelectromechanical systems (MEMS) process to integrate into a lateral flow test strip. The detection of abrin solutions of various concentrations was performed by the as-prepared microfluidic chip. It was shown that the correlation between the abrin concentration and SERS signal was found to be linear within the range of 0.1 ng/mL to 1 μg/mL with a limit of detection of 0.1 ng/mL. Our microfluidic chip design enhanced the operability of SERS-based immunodiagnostic techniques, significantly reducing the complication and cost of preparation as compared to previous SERS-based works. Meanwhile, this design proved the superiority to conventional lateral flow test strips in respect of both sensitivity and quantitation and showed great potential in the diagnosis and treatment for abrin poisoning as well as on-site screening of abrin-spiked materials.

  6. Quality evaluation of yellow peach chips prepared by explosion puffing drying.

    PubMed

    Lyu, Jian; Zhou, Lin-Yan; Bi, Jin-Feng; Liu, Xuan; Wu, Xin-Ye

    2015-12-01

    Nineteen evaluation indicators in 15 yellow peach chips prepared by explosion puffing drying were analyzed, including color, rehydration ratio, texture, and so on. The analysis methods of principle component analysis (PCA), analytic hierarchy process (AHP), K-means cluster (KC) and Discriminate analysis (DA) were used to analyze the comprehensive quality of the yellow peach chips. The dispersed coefficient of variation of the 19 evaluation indicators varied from 3.58 to 852.89 %, suggesting significant differences among yellow peach cultivars. The characteristic evaluation indicators, namely, reducing sugar content, out-put ratio, water content, a value and L value were analyzed by PCA, and their weights 0.0429, 0.1140, 0.4816, 1.1807 and 0.1807 were obtained by AHP. The levels in 15 cultivars effectively were classified by discrimination functions which obtained by KC and DA. The results suggested that three levels of comprehensive quality for yellow peach chips were divided, and the highest synthesis scores was observed in "senggelin" (11.1037), while the lowest synthesis value was found in "goldbaby" (-3.7600).

  7. Rapid identification of ESKAPE bacterial strains using an autonomous microfluidic device.

    PubMed

    Ho, Jack Y; Cira, Nate J; Crooks, John A; Baeza, Josue; Weibel, Douglas B

    2012-01-01

    This article describes Bacteria ID Chips ('BacChips'): an inexpensive, portable, and autonomous microfluidic platform for identifying pathogenic strains of bacteria. BacChips consist of a set of microchambers and channels molded in the elastomeric polymer, poly(dimethylsiloxane) (PDMS). Each microchamber is preloaded with mono-, di-, or trisaccharides and dried. Pressing the layer of PDMS into contact with a glass coverslip forms the device; the footprint of the device in this article is ∼6 cm(2). After assembly, BacChips are degased under large negative pressure and are stored in vacuum-sealed plastic bags. To use the device, the bag is opened, a sample containing bacteria is introduced at the inlet of the device, and the degased PDMS draws the sample into the central channel and chambers. After the liquid at the inlet is consumed, air is drawn into the BacChip via the inlet and provides a physical barrier that separates the liquid samples in adjacent microchambers. A pH indicator is admixed with the samples prior to their loading, enabling the metabolism of the dissolved saccharides in the microchambers to be visualized. Importantly, BacChips operate without external equipment or instruments. By visually detecting the growth of bacteria using ambient light after ∼4 h, we demonstrate that BacChips with ten microchambers containing different saccharides can reproducibly detect the ESKAPE panel of pathogens, including strains of: Enterococcus faecalis, Enteroccocus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter aerogenes, and Enterobacter cloacae. This article describes a BacChip for point-of-care detection of ESKAPE pathogens and a starting point for designing multiplexed assays that identify bacterial strains from clinical samples and simultaneously determine their susceptibility to antibiotics.

  8. A review of microdialysis coupled to microchip electrophoresis for monitoring biological events

    PubMed Central

    Saylor, Rachel A.; Lunte, Susan M.

    2015-01-01

    Microdialysis is a powerful sampling technique that enables monitoring of dynamic processes in vitro and in vivo. The combination of microdialysis with chromatographic or electrophoretic methods yields along with selective detection methods yields a “separation-based sensor” capable of monitoring multiple analytes in near real time. Analysis of microdialysis samples requires techniques that are fast (<1 min), have low volume requirements (nL–pL), and, ideally, can be employed on-line. Microchip electrophoresis fulfills these requirements and also permits the possibility of integrating sample preparation and manipulation with detection strategies directly on-chip. Microdialysis coupled to microchip electrophoresis has been employed for monitoring biological events in vivo and in vitro. This review discusses technical considerations for coupling microdialysis sampling and microchip electrophoresis, including various interface designs, and current applications in the field. PMID:25637011

  9. GaN-based integrated photonics chip with suspended LED and waveguide

    NASA Astrophysics Data System (ADS)

    Li, Xin; Wang, Yongjin; Hane, Kazuhiro; Shi, Zheng; Yan, Jiang

    2018-05-01

    We propose a GaN-based integrated photonics chip with suspended LED and straight waveguide with different geometric parameters. The integrated photonics chip is prepared by double-side process. Light transmission performance of the integrated chip verse current is quantitatively analyzed by capturing light transmitted to waveguide tip and BPM (beam propagation method) simulation. Reduction of the waveguide width from 8 μm to 4 μm results in an over linear reduction of the light output power while a doubling of the length from 250 μm to 500 μm only results in under linear decrease of the output power. Free-space data transmission with 80 Mbps random binary sequence of the integrated chip is capable of achieving high speed data transmission via visible light. This study provides a potential approach for GaN-based integrated photonics chip as micro light source and passive optical device in VLC (visible light communication).

  10. Direct and sensitive detection of foodborne pathogens within fresh produce samples using a field-deployable handheld device.

    PubMed

    You, David J; Geshell, Kenneth J; Yoon, Jeong-Yeol

    2011-10-15

    Direct and sensitive detection of foodborne pathogens from fresh produce samples was accomplished using a handheld lab-on-a-chip device, requiring little to no sample processing and enrichment steps for a near-real-time detection and truly field-deployable device. The detection of Escherichia coli K12 and O157:H7 in iceberg lettuce was achieved utilizing optimized Mie light scatter parameters with a latex particle immunoagglutination assay. The system exhibited good sensitivity, with a limit of detection of 10 CFU mL(-1) and an assay time of <6 min. Minimal pretreatment with no detrimental effects on assay sensitivity and reproducibility was accomplished with a simple and cost-effective KimWipes filter and disposable syringe. Mie simulations were used to determine the optimal parameters (particle size d, wavelength λ, and scatter angle θ) for the assay that maximize light scatter intensity of agglutinated latex microparticles and minimize light scatter intensity of the tissue fragments of iceberg lettuce, which were experimentally validated. This introduces a powerful method for detecting foodborne pathogens in fresh produce and other potential sample matrices. The integration of a multi-channel microfluidic chip allowed for differential detection of the agglutinated particles in the presence of the antigen, revealing a true field-deployable detection system with decreased assay time and improved robustness over comparable benchtop systems. Additionally, two sample preparation methods were evaluated through simulated field studies based on overall sensitivity, protocol complexity, and assay time. Preparation of the plant tissue sample by grinding resulted in a two-fold improvement in scatter intensity over washing, accompanied with a significant increase in assay time: ∼5 min (grinding) versus ∼1 min (washing). Specificity studies demonstrated binding of E. coli O157:H7 EDL933 to only O157:H7 antibody conjugated particles, with no cross-reactivity to K12. This suggests the adaptability of the system for use with a wide variety of pathogens, and the potential to detect in a variety of biological matrices with little to no sample pretreatment. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. A Portable Solid-State Moisture Meter For Agricultural And Food Products

    NASA Astrophysics Data System (ADS)

    Bull, C. R.; Stafford, J. V.; Weaving, G. S.

    1988-10-01

    This paper reports on the development of a small, robust, battery operated near infra-red (NIR) reflectance device, designed for rapid on-farm measurement of the moisture content of forage crops without prior sample preparation. It has potential application to other agricultural or food materials. The instrument is based on two light emitting diodes (LEDs), a germanium detector and a control CMOS single chip microcomputer. The meter has been calibrated to give a direct read out of moisture content for 4 common grass varieties at 3 stages of development. The accuracy of a single point measurement on a grass sample is approximately +/- 6% over a range of 40-80% (wet basis). However, the potential accuracy on a homogeous sample may be as goon as 0.15%.

  12. Paint Analysis

    NASA Technical Reports Server (NTRS)

    1994-01-01

    Lewis Research Center (LEW) has assisted The Cleveland Museum of Art (CMA) in analyzing the museum's paintings. Because of the many layers of paint that are often involved, this is a complex process. The cross-section of a paint chip must be scanned with a microscope to determine whether a paint layer is original or a restoration. The paint samples, however, are rarely flat enough for high magnification viewing and are frequently scratched. LEW devised an automated method that produces intact, flat, polished paint cross-sections. A sophisticated microprocessor-controlled grinding and polishing machine was manually employed in preparation of exotic samples for aerospace research was a readily adaptable technique. It produced perfectly flat samples with clearly defined layers. The process has been used successfully on a number of paintings, and LEW and CMA are considering additional applications.

  13. Integrated electrochemical microsystems for genetic detection of pathogens at the point of care.

    PubMed

    Hsieh, Kuangwen; Ferguson, B Scott; Eisenstein, Michael; Plaxco, Kevin W; Soh, H Tom

    2015-04-21

    The capacity to achieve rapid, sensitive, specific, quantitative, and multiplexed genetic detection of pathogens via a robust, portable, point-of-care platform could transform many diagnostic applications. And while contemporary technologies have yet to effectively achieve this goal, the advent of microfluidics provides a potentially viable approach to this end by enabling the integration of sophisticated multistep biochemical assays (e.g., sample preparation, genetic amplification, and quantitative detection) in a monolithic, portable device from relatively small biological samples. Integrated electrochemical sensors offer a particularly promising solution to genetic detection because they do not require optical instrumentation and are readily compatible with both integrated circuit and microfluidic technologies. Nevertheless, the development of generalizable microfluidic electrochemical platforms that integrate sample preparation and amplification as well as quantitative and multiplexed detection remains a challenging and unsolved technical problem. Recognizing this unmet need, we have developed a series of microfluidic electrochemical DNA sensors that have progressively evolved to encompass each of these critical functionalities. For DNA detection, our platforms employ label-free, single-step, and sequence-specific electrochemical DNA (E-DNA) sensors, in which an electrode-bound, redox-reporter-modified DNA "probe" generates a current change after undergoing a hybridization-induced conformational change. After successfully integrating E-DNA sensors into a microfluidic chip format, we subsequently incorporated on-chip genetic amplification techniques including polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to enable genetic detection at clinically relevant target concentrations. To maximize the potential point-of-care utility of our platforms, we have further integrated sample preparation via immunomagnetic separation, which allowed the detection of influenza virus directly from throat swabs and developed strategies for the multiplexed detection of related bacterial strains from the blood of septic mice. Finally, we developed an alternative electrochemical detection platform based on real-time LAMP, which not is only capable of detecting across a broad dynamic range of target concentrations, but also greatly simplifies quantitative measurement of nucleic acids. These efforts represent considerable progress toward the development of a true sample-in-answer-out platform for genetic detection of pathogens at the point of care. Given the many advantages of these systems, and the growing interest and innovative contributions from researchers in this field, we are optimistic that iterations of these systems will arrive in clinical settings in the foreseeable future.

  14. 5 Steps to Prepare for MACRA Now.

    PubMed

    Martin, Jessica

    2016-10-01

    As the medical community awaits the release of final rules to implement the new Medicare payment system under the Medicare Access and CHIP Reauthorization Act (MACRA), physicians can take steps now to prepare for the transition.

  15. Chip formation and surface integrity in high-speed machining of hardened steel

    NASA Astrophysics Data System (ADS)

    Kishawy, Hossam Eldeen A.

    Increasing demands for high production rates as well as cost reduction have emphasized the potential for the industrial application of hard turning technology during the past few years. Machining instead of grinding hardened steel components reduces the machining sequence, the machining time, and the specific cutting energy. Hard turning Is characterized by the generation of high temperatures, the formation of saw toothed chips, and the high ratio of thrust to tangential cutting force components. Although a large volume of literature exists on hard turning, the change in machined surface physical properties represents a major challenge. Thus, a better understanding of the cutting mechanism in hard turning is still required. In particular, the chip formation process and the surface integrity of the machined surface are important issues which require further research. In this thesis, a mechanistic model for saw toothed chip formation is presented. This model is based on the concept of crack initiation on the free surface of the workpiece. The model presented explains the mechanism of chip formation. In addition, experimental investigation is conducted in order to study the chip morphology. The effect of process parameters, including edge preparation and tool wear on the chip morphology, is studied using Scanning Electron Microscopy (SEM). The dynamics of chip formation are also investigated. The surface integrity of the machined parts is also investigated. This investigation focusses on residual stresses as well as surface and sub-surface deformation. A three dimensional thermo-elasto-plastic finite element model is developed to predict the machining residual stresses. The effect of flank wear is introduced during the analysis. Although residual stresses have complicated origins and are introduced by many factors, in this model only the thermal and mechanical factors are considered. The finite element analysis demonstrates the significant effect of the heat generated during cutting on the residual stresses. The machined specimens are also examined using x-ray diffraction technique to clarify the effect of different speeds, feeds and depths of cut as well as different edge preparations on the residual stress distribution beneath the machined surface. A reasonable agreement between the predicted and measured residual stress is obtained. The results obtained demonstrate the possibility of eliminating the existence of high tensile residual stresses in the workpiece surface by selecting the proper cutting conditions. The machined surfaces are examined using SEM to study the effect of different process parameters and edge preparations on the quality of the machined surface. The phenomenon of material side flow is investigated to clarify the mechanism of this phenomenon. The effect of process parameters and edge preparations on sub-surface deformation is also investigated.

  16. Micron-Scale Differential Scanning Calorimeter on a Chip

    DOEpatents

    Cavicchi, Richard E; Poirier, Gregory Ernest; Suehle, John S; Gaitan, Michael; Tea, Nim H

    1998-06-30

    A differential scanning microcalorimeter produced on a silicon chip enables microscopic scanning calorimetry measurements of small samples and thin films. The chip may be fabricated using standard CMOS processes. The microcalorimeter includes a reference zone and a sample zone. The reference and sample zones may be at opposite ends of a suspended platform or may reside on separate platforms. An integrated polysilicon heater provides heat to each zone. A thermopile consisting of a succession of thermocouple junctions generates a voltage representing the temperature difference between the reference and sample zones. Temperature differences between the zones provide information about the chemical reactions and phase transitions that occur in a sample placed in the sample zone.

  17. Application of IgY to sandwich enzyme-linked immunosorbent assays, lateral flow devices, and immunopillar chips for detecting staphylococcal enterotoxins in milk and dairy products.

    PubMed

    Jin, Wanchun; Yamada, Keiko; Ikami, Mai; Kaji, Noritada; Tokeshi, Manabu; Atsumi, Yusuke; Mizutani, Makoto; Murai, Atsushi; Okamoto, Akira; Namikawa, Takao; Baba, Yoshinobu; Ohta, Michio

    2013-03-01

    Staphylococcal enterotoxins (SEs), produced by Staphylococcus aureus, are a major cause of staphylococcal food poisoning. Traditionally, sandwich enzyme-linked immunosorbent assay (ELISA) and reverse passive latex agglutination with rabbit antibody IgG have been used to detect SEs. However, most of these kits require a long processing time and there is a risk of false-positive results since IgG reacts nonspecifically with protein A produced by S. aureus. In this study, we prepared antienterotoxin chicken IgY antibodies specific for each SE (SEA to SEE) without reaction to protein A, which enabled a drastic reduction in nonspecific reactions. ELISAs, lateral flow device (LFDs), and IgY-based immunopillar chips were developed for SE detection. All the ELISAs developed were as sensitive as commercially available kits. The SEs in milk were successfully detected by the ELISAs, LFDs, and immunopillar chips without any sample pretreatment. The LFD could detect SEA even at the low concentration of 0.2 ng/ml within 15 min in milk. The detection limit of the immunopillar chips for the SEs ranged from 0.01 to 0.1 ng/ml in milk; the SEs were detected within 12 min and specialized skills were not required. The ELISA and LFD detected SEA in dairy products artificially contaminated with S. aureus, including ice cream, yogurt, and café au lait, in a dose-dependent manner. In conclusion, IgY allows highly specific detection of SEs, and ELISAs, LFDs, and immunopillar chips should be useful tools for screening SEs in milk and dairy products. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Microfluidic devices for sample preparation and rapid detection of foodborne pathogens.

    PubMed

    Kant, Krishna; Shahbazi, Mohammad-Ali; Dave, Vivek Priy; Ngo, Tien Anh; Chidambara, Vinayaka Aaydha; Than, Linh Quyen; Bang, Dang Duong; Wolff, Anders

    2018-03-10

    Rapid detection of foodborne pathogens at an early stage is imperative for preventing the outbreak of foodborne diseases, known as serious threats to human health. Conventional bacterial culturing methods for foodborne pathogen detection are time consuming, laborious, and with poor pathogen diagnosis competences. This has prompted researchers to call the current status of detection approaches into question and leverage new technologies for superior pathogen sensing outcomes. Novel strategies mainly rely on incorporating all the steps from sample preparation to detection in miniaturized devices for online monitoring of pathogens with high accuracy and sensitivity in a time-saving and cost effective manner. Lab on chip is a blooming area in diagnosis, which exploits different mechanical and biological techniques to detect very low concentrations of pathogens in food samples. This is achieved through streamlining the sample handling and concentrating procedures, which will subsequently reduce human errors and enhance the accuracy of the sensing methods. Integration of sample preparation techniques into these devices can effectively minimize the impact of complex food matrix on pathogen diagnosis and improve the limit of detections. Integration of pathogen capturing bio-receptors on microfluidic devices is a crucial step, which can facilitate recognition abilities in harsh chemical and physical conditions, offering a great commercial benefit to the food-manufacturing sector. This article reviews recent advances in current state-of-the-art of sample preparation and concentration from food matrices with focus on bacterial capturing methods and sensing technologies, along with their advantages and limitations when integrated into microfluidic devices for online rapid detection of pathogens in foods and food production line. Copyright © 2018. Published by Elsevier Inc.

  19. Applications of Micro/Nanoparticles in Microfluidic Sensors: A Review

    PubMed Central

    Jiang, Yusheng; Wang, Hui; Li, Shunbo; Wen, Weijia

    2014-01-01

    This paper reviews the applications of micro/nanoparticles in microfluidics device fabrication and analytical processing. In general, researchers have focused on two properties of particles—electric behavior and magnetic behavior. The applications of micro/nanoparticles could be summarized on the chip fabrication level and on the processing level. In the fabrication of microfluidic chips (chip fabrication level), particles are good additives in polydimethylsiloxane (PDMS) to prepare conductive or magnetic composites which have wide applications in sensors, valves and actuators. On the other hand, particles could be manipulated according to their electric and magnetic properties under external electric and magnetic fields when they are travelling in microchannels (processing level). Researchers have made a great progress in preparing modified PDMS and investigating the behaviors of particles in microchannels. This article attempts to present a discussion on the basis of particles applications in microfluidics. PMID:24755517

  20. Wood fuel preparation

    Treesearch

    L. H. Reineke

    1965-01-01

    This report gives information on the preparation of wood fuel from wood residues and other wood raw materials. Types of wood fuel discussed are cordwood, stovewood, slabwood, kindling, chips, hogged fuel, sawdust and shavings, bark, charcoal, alcohol, and briquets. Related information is given on types of machinery for preparing wood fuel and on possible markets for...

  1. Visible and near-infrared spectral survey of lunar meteorites recovered by the National Institute of Polar Research

    NASA Astrophysics Data System (ADS)

    Hiroi, T.; Kaiden, H.; Yamaguchi, A.; Kojima, H.; Uemoto, K.; Ohtake, M.; Arai, T.; Sasaki, S.

    2016-12-01

    Lunar meteorite chip samples recovered by the National Institute of Polar Research (NIPR) have been studied by a UV-visible-near-infrared spectrometer, targeting small areas of about 3 × 2 mm in size. Rock types and approximate mineral compositions of studied meteorites have been identified or obtained through this spectral survey with no sample preparation required. A linear deconvolution method was used to derive end-member mineral spectra from spectra of multiple clasts whenever possible. In addition, the modified Gaussian model was used in an attempt of deriving their major pyroxene compositions. This study demonstrates that a visible-near-infrared spectrometer on a lunar rover would be useful for identifying these kinds of unaltered (non-space-weathered) lunar rocks. In order to prepare for such a future mission, further studies which utilize a smaller spot size are desired for improving the accuracy of identifying the clasts and mineral phases of the rocks.

  2. Enrichment of wheat chips with omega-3 fatty acid by flaxseed addition: textural and some physicochemical properties.

    PubMed

    Yuksel, Ferhat; Karaman, Safa; Kayacier, Ahmed

    2014-02-15

    In the present study, wheat chips enriched with flaxseed flour were produced and response surface methodology was used for the studying the simultaneous effects of flaxseed level (10-20%), frying temperature (160-180 °C) and frying time (40-60 s) on some physicochemical, textural and sensorial properties and fatty acid composition of wheat chips. Ridge analysis was conducted to determine the optimum levels of processing variables. Predictive regression equations with adequate coefficients of determination (R² ≥ 0.705) to explain the effect of processing variables were constructed. Addition of flaxseed flour increased the dry matter and protein content of samples and increase of frying temperature decreased the hardness values of wheat chips samples. Increment in flaxseed level provided an increase in unsaturated fatty acid content namely omega-3 fatty acids of wheat chips samples. Overall acceptability of chips increased with the increase of frying temperature. Ridge analysis showed that maximum taste score would be at flaxseed level = 10%, frying temperature = 180 °C and frying time = 50 s. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. 40 Years of Processing Pieces of Space

    NASA Technical Reports Server (NTRS)

    Satterwhite, C. E.; Funk, R. C.; Righter, K.; Harrington, R. H.

    2016-01-01

    This year marks the 40th year anniversary for the Antarctic Search for Meteorite (ANSMET) program. In 1976, the ANSMET program led the first expedition to Antarctica. The ANSMET program is a US-led field-based science project that recovers meteorite samples from Antarctica. Once a year from late November to late January, a field team consisting of 8 to 12 people, spends 6-8 weeks camping on the ice and collecting meteorites. Since 1976, more than 22,000 meteorite samples have been recovered. These meteorites come from asteroids, planets and other bodies of the solar system. Once collected, the Antarctic meteorites are shipped to NASA/Johnson Space Center (JSC) Houston, TX. in a refrigerated truck and are kept frozen to minimize oxidation until they are ready for initial processing. In Antarctica each meteorite is given a field tag which consists of numbers, once in the lab, this is replaced by an official tag, consisting of the Antarctic field location and year collected. The types and numbers of meteorites that have been classified include 849 carbonaceous chondrites, 135 enstatites, 512 achondrites, 64 stony, 115 irons, 48 others (27 R chondrites, 7 ungrouped), 6,161 H chondrites, 7,668 L chondrites, and 4,589 LL chondrites. Although 80-85 percent of the collected meteorites fall in the ordinary chondrite group, the other approximately 15 percent represent rare types of achondrites and carbonaceous chondrites. These rare meteorites include 25 lunar meteorites, 15 Martian meteorites, scores of various types of carbonaceous chondrites, and unique achondrites. The Antarctic meteorites that have been collected are processed in the Meteorite Processing Lab at JSC in Houston, TX. Initial processing of the meteorites begins with thawing/drying the meteorites in a nitrogen glove box for 24 to 48 hours. The meteorites are then photographed, measured, weighed and a description of the interior and exterior of each meteorite is written. The meteorite is broken and a representative sample, either a 1-3 gram chip or thin section is sent to the Smithsonian Institution for classification. After Antarctic meteorites have been classified and approved by the Nomenclature Committee of the Meteoritical Society, they are announced in the Antarctic Meteorite Newsletter, which is published twice per year (fall and spring) so that scientists may review which meteorites are available to study. Requests for Antarctic Meteorite samples are welcomed from research scientists, regardless of their current state of funding for meteorite studies. Since its inception over 3,300 requests have been made for pieces of these meteorites and over 400 investigators worldwide are active in the study of meteorites.. Research on these samples has been published in more than1500 peer reviewed articles; a listing of papers for any meteorite sample can be generated by accessing http://curator.jsc.nasa.gov/antmet/referencesearch.cfm. Antarctic meteorite samples requested by scientists are prepared several different ways. Most samples are prepared as chips, either using a rock splitter or using a chisel and chipping bowl. In special situations, a researcher may request a meteorite slab in which case the samples are cut using a diamond-bladed bandsaw inside of a dry nitrogen glove box. The meteorites are always cut in a 100 percent liquid-free environment. Additionally, thin/thick sections of Antarctic meteorites are also prepared at JSC. The meteorite thin section lab at JSC can prepare standard 30-micron thin sections, thick sections of variable thickness (100 to 200 microns), or demountable sections using superglue, all section are prepared without using water. Although many of the techniques used back in the '70's are still used today, advances in computers, software, databases, available tools and instrumentation have helped to streamline and shorten the duration of the classification process. In conjunction with present day missions to asteroids and other planets, meteorite studies have not only led to a better understanding of the complex histories of these bodies but have also tied certain meteorite groups to particular asteroid bodies. New meteorite discoveries by the ANSMET program provide a cost effective method for obtaining samples of previously unsampled bodies, allowing scientists to learn more about the origin, composition, and evolution of the solar system. Preservation in our cleanrooms at NASA allows material to be archived for future generations and advances in instrumentation and analysis.

  4. Next Generation Instrumentation Bus Test Plan for Fibre Channel

    DTIC Science & Technology

    1999-09-30

    sample for port testing. The heart of the Fibre Xpress network cards is the tachyon chip from Hewlett Packard. The tachyon chip is basically a single...be to test the protocols. 1.5.1.1 Tachyon chip The user’s manual for the Tachyon controller chip identifies the following FC-AL specification

  5. [Preparation of poly(methyl acrylate) microfluidic chips surface-modified by hyperbranched polyamide ester and their application in the separation of biomolecules].

    PubMed

    Liu, Bing; Lin, Donge; Xu, Lin; Lei, Yanhui; Bo, Qianglong; Shou, Chongqi

    2012-05-01

    The surface of poly (methyl acrylate) (PMMA) microfluidic chips were modified using hyperbranched polyamide ester via chemical bonding. The contact angles of the modified chips were measured. The surface morphology was observed by scanning electron microscope (SEM) and stereo microscope. The results showed that the surface of the modified chips was coated by a dense, uniform, continuous, hydrophilic layer of hyperbranched polyamide ester. The hydrophilic of the chip surface was markedly improved. The contact angle of the chips modified decreased from 89.9 degrees to 29.5 degrees. The electro osmotic flow (EOF) in the modified microchannel was lower than that in the unmodified microchannel. Adenosine and L-lysine were detected and separated via the modified PMMA microfluidic chips. Compared with unmodified chips, the modified chips successfully separated the two biomolecules. The detection peaks were clear and sharp. The separation efficiencies of adenosine and L-lysine were 8.44 x 10(4) plates/m and 9.82 x 10(4) plates/m respectively, and the resolutions (Rs) was 5.31. The column efficiencies and resolutions of the modified chips were much higher than those of the unmodified chips. It was also observed that the modified chips possessed good reproducibility of migration time. This research may provide a new and effective method to improve the hydrophilicity of the PMMA surface and the application of PMMA microfluidic chips in the determination of trace biomolecules.

  6. Structural elucidation and magnetic behavior evaluation of Cu-Cr doped BaCo-X hexagonal ferrites

    NASA Astrophysics Data System (ADS)

    Azhar Khan, Muhammad; Hussain, Farhat; Rashid, Muhammad; Mahmood, Asif; Ramay, Shahid M.; Majeed, Abdul

    2018-04-01

    Ba2-xCuxCo2CryFe28-yO46 (x = 0.0, 0.1, 0.2, 0.3, 0.4, y = 0.0, 0.2, 0.4, 0.6, 0.8) X-type hexagonal ferrites were synthesized via micro-emulsion route. The techniques which were applied to characterize the prepared samples are as follows: X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Dielectric measurements and vibrating sample magnetometer (VSM). The structural parameters i.e. lattice constant (a, c), cell volume (V), X-ray density, bulk density and crystallite size of all the prepared samples were obtained using XRD analysis. The lattice parameters 'a' and 'c' increase from 5.875 Å to 5.934 Å and 83.367 Å to 83.990 Å respectively. The crystallite size of investigated samples lies in the range of 28-32 nm. The magnetic properties of all samples have been calculated by vibrating sample magnetometer (VSM) analysis. The increase in coercivity (Hc) was observed with the increase of doping contents. It was observed that the coercivity (Hc) of all prepared samples is inversely related to the crystalline size which reflects that all materials are super-paramagnetic. The dielectric parameters i.e. dielectric constant, dielectric loss, tangent loss etc were obtained in the frequency range of 1 MHz-3 GHz and followed the Maxwell-Wagner's model. The significant variation the dielectric parameters are observed with increasing frequency. The maximum Q value is obtained at ∼2 GHz due to which these materials are used for high frequency multilayer chip inductors.

  7. Study of temperature rises and forces on drilling bone

    NASA Astrophysics Data System (ADS)

    Srikanth Venkataraman, Ananya

    Many different approaches have been used to prepare, store and test bone samples in order to determine its physical properties. The need to establish a standard method of specimen preparation and storage prior to experimental testing, contributed greatly to the primary part of this study. When mechanized cutting tools such as saws and drills are used, heat is produced and this raises the temperature of both the tool and the material being cut. In orthopedic and dental practices, high-speed tools are often applied to bones and teeth, and heat from these operations may result in thermal necrosis [1]. Since this can have a negative impact on the outcome of an orthopedic procedure, temperatures must be kept below the threshold that results in bone necrosis. The initial set of experiments was performed to determine the conditions under which the mechanical properties of the bone changed so as to establish the most suitable testing conditions. The hardness variation of the bone samples, under different annealing treatment conditions was used as the indicating parameter for evaluation of the change in the mechanical properties. Establishing the most appropriate section of the metacarpal sample for testing, by studying the anisotropy of the bone was another determining parameter. The second step was to examine the effects of conventional drilling as well as modulation assisted drilling on the temperature rise generated in the bone during these machining processes. In addition to this, a set of experiments were performed to ascertain how lubrication affected the temperature rise during drilling. The dynamic portions of the torque and thrust traces as well as the specific energies were compared for the different drilling conditions. Modulation showed no significant effect on the mean torque, thrust, specific energies of cutting, or temperature rise. Lubrication (flooding and misting) in both the modulation and no modulation cases drastically reduced the temperature rise during cutting, as expected. In addition to this the characteristics of the chips produced by both the methods of drilling were compared. The modulation process produced more consistent chips at the lower speed (360 rpm) and as the speed was increased to 3000 rpm the chip formation was similar to the no modulation drilling condition at the same feed rates. A brief study on the histological changes due to drilling was also performed.

  8. Electrical lysis: dynamics revisited and advances in On-chip operation.

    PubMed

    Morshed, Bashir; Shams, Maitham; Mussivand, Tofy

    2013-01-01

    Electrical lysis (EL) is the process of breaking the cell membrane to expose the internal contents under an applied high electric field. Lysis is an important phenomenon for cellular analysis, medical treatment, and biofouling control. This paper aims to review, summarize, and analyze recent advancements on EL. Major databases including PubMed, Ei Engineering Village, IEEE Xplore, and Scholars Portal were searched using relevant keywords. More than 50 articles published in English since 1997 are cited in this article. EL has several key advantages compared to other lysis techniques such as chemical, mechanical, sonication, or laser, including rapid speed of operation, ability to control, miniaturization, low cost, and low power requirement. A variety of cell types have been investigated for including protoplasts, E. coli, yeasts, blood cells, and cancer cells. EL has been developed and applied for decontamination, cytology, genetics, single-cell analysis, cancer treatment, and other applications. On-chip EL is a promising technology for multiplexed automated implementation of cell-sample preparation and processing with micro- or nanoliter reagents.

  9. Detection of acrylamide in potato chips using a fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots.

    PubMed

    Hu, Qinqin; Xu, Xiahong; Li, Zhanming; Zhang, Ying; Wang, Jianping; Fu, Yingchun; Li, Yanbin

    2014-04-15

    Acrylamide is a neurotoxin and potential carcinogen, but is found in various thermally processed foods such as potato chips, biscuits, and coffee. Simple and sensitive methods for on-line detection of acrylamide are needed to ensure food safety. In this paper, a novel fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots (QDs) was proposed for detecting acrylamide in potato chips. The functional QDs were prepared by their binding with N-acryloxysuccinimide (NAS), which was characterized by Fourier transform infrared (FR-IR) spectra. The carbon-carbon double bonds of NAS modified QDs polymerized with assistance of photo initiator under UV irradiation, leading to QDs getting closer along with fluorescence intensity decreasing. Acrylamide in the sample participated in the polymerization and induced an increase of fluorescence intensity. This method possessed a linear range from 3.5×10(-5) to 3.5 g L(-1) (r(2)=0.94) and a limit of detection of 3.5×10(-5) g L(-1). Although the sensitivity and specificity cannot be compared with standard LC-MS/MS analysis, this new method requires much less time and cost, which is promising for on-line rapid detection of acrylamide in food processing. © 2013 Published by Elsevier B.V.

  10. Strategies for comparing gene expression profiles from different microarray platforms: application to a case-control experiment.

    PubMed

    Severgnini, Marco; Bicciato, Silvio; Mangano, Eleonora; Scarlatti, Francesca; Mezzelani, Alessandra; Mattioli, Michela; Ghidoni, Riccardo; Peano, Clelia; Bonnal, Raoul; Viti, Federica; Milanesi, Luciano; De Bellis, Gianluca; Battaglia, Cristina

    2006-06-01

    Meta-analysis of microarray data is increasingly important, considering both the availability of multiple platforms using disparate technologies and the accumulation in public repositories of data sets from different laboratories. We addressed the issue of comparing gene expression profiles from two microarray platforms by devising a standardized investigative strategy. We tested this procedure by studying MDA-MB-231 cells, which undergo apoptosis on treatment with resveratrol. Gene expression profiles were obtained using high-density, short-oligonucleotide, single-color microarray platforms: GeneChip (Affymetrix) and CodeLink (Amersham). Interplatform analyses were carried out on 8414 common transcripts represented on both platforms, as identified by LocusLink ID, representing 70.8% and 88.6% of annotated GeneChip and CodeLink features, respectively. We identified 105 differentially expressed genes (DEGs) on CodeLink and 42 DEGs on GeneChip. Among them, only 9 DEGs were commonly identified by both platforms. Multiple analyses (BLAST alignment of probes with target sequences, gene ontology, literature mining, and quantitative real-time PCR) permitted us to investigate the factors contributing to the generation of platform-dependent results in single-color microarray experiments. An effective approach to cross-platform comparison involves microarrays of similar technologies, samples prepared by identical methods, and a standardized battery of bioinformatic and statistical analyses.

  11. Portable microfluidic platform for real-time, high sensitive detection and identification of trichloroethylene and other organochloride compounds

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jensen, Erik

    In this successful SBIR Phase II effort, HJ Science & Technology, Inc. has designed and built a novel portable instrument capable of performing automated aqueous organochloride (chlorinated solvent) speciation analysis for environmental monitoring at DoE sites. Our technique employs performing organochloride conjugation, labeling the conjugate with an efficient fluorophore, and performing on-chip capillary electrophoresis separation with laser induced fluorescence detection. The key component of the portable instrument is a novel microfluidic chip capable of complete “end-to-end” automation of sample preparation, conjugation, labeling, and μCE separation and detection. In addition, the Phase II prototype includes key supporting instrumentation such as themore » optical module, pneumatic manifold, electronics, software, etc. As such, we have achieved all of the following 4 Phase II technical objectives: 1) Further refine and optimize the “on-chip” automation of the organochloride conjugation and labeling protocol, 2) Further improve the microfluidic chip fabrication process and the pneumatic manifold design in order to address issues related to performance consistency, product yield, performance reliability, and user friendliness, 3) Design and build the supporting components of the Phase II prototype including optical module, electronics, and software, and 4) Assemble the Phase II prototype hardware.« less

  12. Fabrication of Smart Chemical Sensors Based on Transition-Doped-Semiconductor Nanostructure Materials with µ-Chips

    PubMed Central

    Rahman, Mohammed M.; Khan, Sher Bahadar; Asiri, Abdullah M.

    2014-01-01

    Transition metal doped semiconductor nanostructure materials (Sb2O3 doped ZnO microflowers, MFs) are deposited onto tiny µ-chip (surface area, ∼0.02217 cm2) to fabricate a smart chemical sensor for toxic ethanol in phosphate buffer solution (0.1 M PBS). The fabricated chemi-sensor is also exhibited higher sensitivity, large-dynamic concentration ranges, long-term stability, and improved electrochemical performances towards ethanol. The calibration plot is linear (r2 = 0.9989) over the large ethanol concentration ranges (0.17 mM to 0.85 M). The sensitivity and detection limit is ∼5.845 µAcm−2mM−1 and ∼0.11±0.02 mM (signal-to-noise ratio, at a SNR of 3) respectively. Here, doped MFs are prepared by a wet-chemical process using reducing agents in alkaline medium, which characterized by UV/vis., FT-IR, Raman, X-ray photoelectron spectroscopy (XPS), powder X-ray diffraction (XRD), and field-emission scanning electron microscopy (FE-SEM) etc. The fabricated ethanol chemical sensor using Sb2O3-ZnO MFs is simple, reliable, low-sample volume (<70.0 µL), easy of integration, high sensitivity, and excellent stability for the fabrication of efficient I–V sensors on μ-chips. PMID:24454785

  13. Fabrication of smart chemical sensors based on transition-doped-semiconductor nanostructure materials with µ-chips.

    PubMed

    Rahman, Mohammed M; Khan, Sher Bahadar; Asiri, Abdullah M

    2014-01-01

    Transition metal doped semiconductor nanostructure materials (Sb2O3 doped ZnO microflowers, MFs) are deposited onto tiny µ-chip (surface area, ∼0.02217 cm(2)) to fabricate a smart chemical sensor for toxic ethanol in phosphate buffer solution (0.1 M PBS). The fabricated chemi-sensor is also exhibited higher sensitivity, large-dynamic concentration ranges, long-term stability, and improved electrochemical performances towards ethanol. The calibration plot is linear (r(2) = 0.9989) over the large ethanol concentration ranges (0.17 mM to 0.85 M). The sensitivity and detection limit is ∼5.845 µAcm(-2)mM(-1) and ∼0.11±0.02 mM (signal-to-noise ratio, at a SNR of 3) respectively. Here, doped MFs are prepared by a wet-chemical process using reducing agents in alkaline medium, which characterized by UV/vis., FT-IR, Raman, X-ray photoelectron spectroscopy (XPS), powder X-ray diffraction (XRD), and field-emission scanning electron microscopy (FE-SEM) etc. The fabricated ethanol chemical sensor using Sb2O3-ZnO MFs is simple, reliable, low-sample volume (<70.0 µL), easy of integration, high sensitivity, and excellent stability for the fabrication of efficient I-V sensors on μ-chips.

  14. Fabrication Localized Surface Plasmon Resonance sensor chip of gold nanoparticles and detection lipase-osmolytes interaction

    NASA Astrophysics Data System (ADS)

    Ghodselahi, T.; Hoornam, S.; Vesaghi, M. A.; Ranjbar, B.; Azizi, A.; Mobasheri, H.

    2014-09-01

    Co-deposition of RF-sputtering and RF-PECVD from acetylene gas and Au target were used to prepare sensor chip of gold nanoparticles (Au NPs). Deposition conditions were optimized to reach a Localized Surface Plasmon Resonance (LSPR) sensor chip of Au NPs with particle size less than 10 nm. The RF power was set at 180 W and the initial gas pressure was set at 0.035 mbar. Transmission Electron Microscopy (TEM) images and Atomic Force Microscopy (AFM) data were used to investigate particles size and surface morphology of LSPR sensor chip. The Au and C content of the LSPR sensor chip of Au NPs was obtained from X-ray photoelectron spectroscopy (XPS). The hydrogenated amorphous carbon (a-C:H) thin film was used as intermediate material to immobilize Au NPs on the SiO2 substrate. The interaction between two types of osmolytes, i.e. sorbitol and trehalose, with Pseudomonas cepacia lipase (PCL) were detected by the prepared LSPR biosensor chip. The detection mechanism is based on LSPR spectroscopy in which the wavelength of absorption peak is sensitive to the refractive index of the environment of the Au NPs. This mechanism eliminates the use of a probe or immobilization of PCL on the Au NPs of LSPR sensor chip. The interaction between PCL and osmolytes can change refractive index of the mixture or solution. We found that unlike to trehalose, sorbitol interacts with the PCL. This interaction increases refractive index of the PCL and sorbitol mixture. Refractive index of PCL in the presence of different concentration of sorbitol was obtained by Mie theory modeling of LSPR peaks. This modeling stated that the present LSPR sensor chip has sensitivity as high as wavelength shift of 175 nm per refractive index. Moreover, the detection of such weakly interaction between bio-molecules cannot be achieved by other analysis.

  15. Silicon sample holder for molecular beam epitaxy on pre-fabricated integrated circuits

    NASA Technical Reports Server (NTRS)

    Hoenk, Michael E. (Inventor); Grunthaner, Paula J. (Inventor); Grunthaner, Frank J. (Inventor)

    1994-01-01

    The sample holder of the invention is formed of the same semiconductor crystal as the integrated circuit on which the molecular beam expitaxial process is to be performed. In the preferred embodiment, the sample holder comprises three stacked micro-machined silicon wafers: a silicon base wafer having a square micro-machined center opening corresponding in size and shape to the active area of a CCD imager chip, a silicon center wafer micro-machined as an annulus having radially inwardly pointing fingers whose ends abut the edges of and center the CCD imager chip within the annulus, and a silicon top wafer micro-machined as an annulus having cantilevered membranes which extend over the top of the CCD imager chip. The micro-machined silicon wafers are stacked in the order given above with the CCD imager chip centered in the center wafer and sandwiched between the base and top wafers. The thickness of the center wafer is about 20% less than the thickness of the CCD imager chip. Preferably, four titanium wires, each grasping the edges of the top and base wafers, compress all three wafers together, flexing the cantilever fingers of the top wafer to accommodate the thickness of the CCD imager chip, acting as a spring holding the CCD imager chip in place.

  16. Density determination of nail polishes and paint chips using magnetic levitation

    NASA Astrophysics Data System (ADS)

    Huang, Peggy P.

    Trace evidence is often small, easily overlooked, and difficult to analyze. This study describes a nondestructive method to separate and accurately determine the density of trace evidence samples, specifically nail polish and paint chip using magnetic levitation (MagLev). By determining the levitation height of each sample in the MagLev device, the density of the sample is back extrapolated using a standard density bead linear regression line. The results show that MagLev distinguishes among eight clear nail polishes, including samples from the same manufacturer; separates select colored nail polishes from the same manufacturer; can determine the density range of household paint chips; and shows limited levitation for unknown paint chips. MagLev provides a simple, affordable, and nondestructive means of determining density. The addition of co-solutes to the paramagnetic solution to expand the density range may result in greater discriminatory power and separation and lead to further applications of this technique.

  17. A microfluidic sub-critical water extraction instrument

    NASA Astrophysics Data System (ADS)

    Sherrit, Stewart; Noell, Aaron C.; Fisher, Anita; Lee, Mike C.; Takano, Nobuyuki; Bao, Xiaoqi; Kutzer, Thomas C.; Grunthaner, Frank

    2017-11-01

    This article discusses a microfluidic subcritical water extraction (SCWE) chip for autonomous extraction of amino acids from astrobiologically interesting samples. The microfluidic instrument is composed of three major components. These include a mixing chamber where the soil sample is mixed and agitated with the solvent (water), a subcritical water extraction chamber where the sample is sealed with a freeze valve at the chip inlet after a vapor bubble is injected into the inlet channels to ensure the pressure in the chip is in equilibrium with the vapor pressure and the slurry is then heated to ≤200 °C in the SCWE chamber, and a filter or settling chamber where the slurry is pumped to after extraction. The extraction yield of the microfluidic SCWE chip process ranged from 50% compared to acid hydrolysis and 80%-100% compared to a benchtop microwave SCWE for low biomass samples.

  18. Droplet-Based Segregation and Extraction of Concentrated Samples

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Buie, C R; Buckley, P; Hamilton, J

    2007-02-23

    Microfluidic analysis often requires sample concentration and separation techniques to isolate and detect analytes of interest. Complex or scarce samples may also require an orthogonal separation and detection method or off-chip analysis to confirm results. To perform these additional steps, the concentrated sample plug must be extracted from the primary microfluidic channel with minimal sample loss and dilution. We investigated two extraction techniques; injection of immiscible fluid droplets into the sample stream (''capping'''') and injection of the sample into an immiscible fluid stream (''extraction''). From our results we conclude that capping is the more effective partitioning technique. Furthermore, this functionalitymore » enables additional off-chip post-processing procedures such as DNA/RNA microarray analysis, realtime polymerase chain reaction (RT-PCR), and culture growth to validate chip performance.« less

  19. Quantitative Investigation of Protein-Nucleic Acid Interactions by Biosensor Surface Plasmon Resonance.

    PubMed

    Wang, Shuo; Poon, Gregory M K; Wilson, W David

    2015-01-01

    Biosensor-surface plasmon resonance (SPR) technology has emerged as a powerful label-free approach for the study of nucleic acid interactions in real time. The method provides simultaneous equilibrium and kinetic characterization for biomolecular interactions with low sample requirements and without the need for external probes. A detailed and practical guide for protein-DNA interaction analyses using biosensor-SPR methods is presented. Details of SPR technology and basic fundamentals are described with recommendations on the preparation of the SPR instrument, sensor chips and samples, experimental design, quantitative and qualitative data analyses and presentation. A specific example of the interaction of a transcription factor with DNA is provided with results evaluated by both kinetic and steady-state SPR methods.

  20. [GSTP1, APC and RASSF1 gene methylation in prostate cancer samples: comparative analysis of MS-HRM method and Infinium HumanMethylation450 BeadChip beadchiparray diagnostic value].

    PubMed

    Skorodumova, L O; Babalyan, K A; Sultanov, R; Vasiliev, A O; Govorov, A V; Pushkar, D Y; Prilepskaya, E A; Danilenko, S A; Generozov, E V; Larin, A K; Kostryukova, E S; Sharova, E I

    2016-11-01

    There is a clear need in molecular markers for prostate cancer (PC) risk stratification. Alteration of DNA methylation is one of processes that occur during ÐÑ progression. Methylation-sensitive PCR with high resolution melting curve analysis (MS-HRM) can be used for gene methylation analysis in routine laboratory practice. This method requires very small amounts of DNA for analysis. Numerous results have been accumulated on DNA methylation in PC samples analyzed by the Infinium HumanMethylation450 BeadChip (HM450). However, the consistency of MS-HRM results with chip hybridization results has not been examined yet. The aim of this study was to assess the consistency of results of GSTP1, APC and RASSF1 gene methylation analysis in ÐÑ biopsy samples obtained by MS-HRM and chip hybridization. The methylation levels of each gene determined by MS-HRM were statistically different in the group of PC tissue samples and the samples without signs of tumor growth. Chip hybridization data analysis confirmed the results obtained with the MS-HRM. Differences in methylation levels between tumor tissue and histologically intact tissue of each sample determined by MS-HRM and chip hybridization, were consistent with each other. Thus, we showed that the assessment of GSTP1, APC and RASSF1 gene methylation analysis using MS-HRM is suitable for the design of laboratory assays that will differentiate the PC tissue from the tissue without signs of tumor growth.

  1. Digital Microfluidics Sample Analyzer

    NASA Technical Reports Server (NTRS)

    Pollack, Michael G.; Srinivasan, Vijay; Eckhardt, Allen; Paik, Philip Y.; Sudarsan, Arjun; Shenderov, Alex; Hua, Zhishan; Pamula, Vamsee K.

    2010-01-01

    Three innovations address the needs of the medical world with regard to microfluidic manipulation and testing of physiological samples in ways that can benefit point-of-care needs for patients such as premature infants, for which drawing of blood for continuous tests can be life-threatening in their own right, and for expedited results. A chip with sample injection elements, reservoirs (and waste), droplet formation structures, fluidic pathways, mixing areas, and optical detection sites, was fabricated to test the various components of the microfluidic platform, both individually and in integrated fashion. The droplet control system permits a user to control droplet microactuator system functions, such as droplet operations and detector operations. Also, the programming system allows a user to develop software routines for controlling droplet microactuator system functions, such as droplet operations and detector operations. A chip is incorporated into the system with a controller, a detector, input and output devices, and software. A novel filler fluid formulation is used for the transport of droplets with high protein concentrations. Novel assemblies for detection of photons from an on-chip droplet are present, as well as novel systems for conducting various assays, such as immunoassays and PCR (polymerase chain reaction). The lab-on-a-chip (a.k.a., lab-on-a-printed-circuit board) processes physiological samples and comprises a system for automated, multi-analyte measurements using sub-microliter samples of human serum. The invention also relates to a diagnostic chip and system including the chip that performs many of the routine operations of a central labbased chemistry analyzer, integrating, for example, colorimetric assays (e.g., for proteins), chemiluminescence/fluorescence assays (e.g., for enzymes, electrolytes, and gases), and/or conductometric assays (e.g., for hematocrit on plasma and whole blood) on a single chip platform.

  2. Rapid Identification of ESKAPE Bacterial Strains Using an Autonomous Microfluidic Device

    PubMed Central

    Ho, Jack Y.; Cira, Nate J.; Crooks, John A.; Baeza, Josue; Weibel, Douglas B.

    2012-01-01

    This article describes Bacteria ID Chips (‘BacChips’): an inexpensive, portable, and autonomous microfluidic platform for identifying pathogenic strains of bacteria. BacChips consist of a set of microchambers and channels molded in the elastomeric polymer, poly(dimethylsiloxane) (PDMS). Each microchamber is preloaded with mono-, di-, or trisaccharides and dried. Pressing the layer of PDMS into contact with a glass coverslip forms the device; the footprint of the device in this article is ∼6 cm2. After assembly, BacChips are degased under large negative pressure and are stored in vacuum-sealed plastic bags. To use the device, the bag is opened, a sample containing bacteria is introduced at the inlet of the device, and the degased PDMS draws the sample into the central channel and chambers. After the liquid at the inlet is consumed, air is drawn into the BacChip via the inlet and provides a physical barrier that separates the liquid samples in adjacent microchambers. A pH indicator is admixed with the samples prior to their loading, enabling the metabolism of the dissolved saccharides in the microchambers to be visualized. Importantly, BacChips operate without external equipment or instruments. By visually detecting the growth of bacteria using ambient light after ∼4 h, we demonstrate that BacChips with ten microchambers containing different saccharides can reproducibly detect the ESKAPE panel of pathogens, including strains of: Enterococcus faecalis, Enteroccocus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter aerogenes, and Enterobacter cloacae. This article describes a BacChip for point-of-care detection of ESKAPE pathogens and a starting point for designing multiplexed assays that identify bacterial strains from clinical samples and simultaneously determine their susceptibility to antibiotics. PMID:22848451

  3. On-chip micropatterning of plastic (cylic olefin copolymer, COC) microfluidic channels for the fabrication of biomolecule microarrays using photografting methods.

    PubMed

    Pu, Qiaosheng; Oyesanya, Olufemi; Thompson, Bowlin; Liu, Shantang; Alvarez, Julio C

    2007-01-30

    This paper reports on the surface modification of plastic microfluidic channels to prepare different biomolecule micropatterns using ultraviolet (UV) photografting methods. The linkage chemistry is based upon UV photopolymerization of acryl monomers to generate thin films (0.01-6 microm) chemically linked to the organic backbone of the plastic surface. The commodity thermoplastic, cyclic olefin copolymer (COC) was selected to build microfluidic chips because of its significant UV transparency and easiness for microfabrication by molding techniques. Once the polyacrylic films were grafted on the COC surface using photomasks, micropatterns of proteins, DNA, and biotinlated conjugates were readily obtained by surface chemical reactions in one or two subsequent steps. The thickness of the photografted films can be tuned from several nanometers up to several micrometers, depending on the reaction conditions. The micropatterned films can be prepared inside the microfluidic channel (on-chip) or on open COC surfaces (off-chip) with densities of functional groups about 10(-7) mol/cm2. Characterization of these films was performed by attenuated-total-reflectance IR spectroscopy, fluorescence microscopy, profilometry, atomic force microscopy, and electrokinetic methods.

  4. Morphologies and optical and electrical properties of InGaN/GaN micro-square array light-emitting diode chips.

    PubMed

    Han, Dan; Ma, Shufang; Jia, Zhigang; Liu, Peizhi; Jia, Wei; Shang, Lin; Zhai, Guangmei; Xu, Bingshe

    2018-04-10

    InGaN/GaN micro-square array light-emitting diode (LED) chips (micro-chips) have been prepared via the focused ion beam (FIB) etching technique, which can not only reduce ohmic contact degradation but also control the aspect ratio precisely in three-dimensional (3D) structure LED (3D-LED) device fabrication. The effects of FIB beam current and micro-square array depth on morphologies and optical and electrical properties of the micro-chips have been studied. Our results show that sidewall surface morphology and optical and electrical properties of the micro-chips degrade with increased beam current. After potassium hydroxide etching with different times, an optimal current-voltage and luminescence performance can be obtained. Combining the results of cathodoluminescence mappings and light output-current characteristics, the light extraction efficiency of the micro-chips is reduced as FIB etch depth increases. The mechanisms of micro-square depth on light extraction have been revealed by 3D finite difference time domain.

  5. Development of an anti-EPO antibody detection kit based on lab-on-a-chip and bridging antibody technologies.

    PubMed

    Oh, Jin-Gyo; Seong, Jihyun; Han, Sunmi; Heo, Tae-Hwe

    2018-05-17

    Immunogenicity is a major concern in the use of biological drugs. In particular, antibody-mediated pure red cell aplasia (PRCA) is a rare condition that is caused by administration of recombinant erythropoietin. There are numerous assay platforms for detect EPO anti-drug antibody (ADA), and most have appropriate assay sensitivity, but in need of improvement in terms of assay turnaround time and user accessibility. Here, the new method was developed based on lab-on-a-chip technology and bridging ELISA. The FREND™ Cartridge is equipped with a microfluidic lateral flow channel, enabling easy, fast and accurate immunoassays with small sample volumes. Biotinylated EPO was immobilized on the avidin-coated solid phase of the test zone in the FREND™ cartridge. Initially, ADA in the serum sample binds to the detector conjugate (EPO-HRP-anti HRP antibody-FL bead) in the conjugation zone, and it flows into the test zone prepared with capture complex (avidin-biotinylated EPO). Unbound detector complexes are captured in the reference zone. The FREND™ system detects and quantifies the fluorescence signals in each zone and then calculates the concentration of EPO ADA in the sample. The FREND™ EPO ADA kit may be useful in local clinics as a rapid method for monitoring patients administered recombinant erythropoietin. Copyright © 2018 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  6. Stress insensitive multilayer chip inductor with ferrite core

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Vishwas, B.; Madhuri, W., E-mail: madhuriw12@gmail.com; Rao, N. Madhusudan

    2015-06-24

    Mg{sub 0.25}Cu{sub 0.25}Zn{sub 0.5}Fe{sub 2}O{sub 4} is synthesized by sol gel auto combustion technique. The obtained ferrite powder is finally sintered in a microwave furnace at 850°C. Multilayer chip inductor (MLCI) of two layers is prepared by screen printing technique. The sintered ferrite is characterized by X-ray diffraction. The frequency response of dielectric constant is studied in the frequency range of 100Hz to 5MHz. Dielectric polarization is discussed in the light of Maxwell-Wagner interfacial polarization. The prepared MLCI is studied for stress sensitivity in the range of 0 to 8 MPa.

  7. Ignition propagation and heat effects of propellant chips embedded in castable inhibitor using a laser flux test bomb

    NASA Technical Reports Server (NTRS)

    Bolton, Douglas E., Jr.

    1993-01-01

    A castable inhibitor is applied to the aft face of the Space Shuttle Redesigned Solid Rocket Motor (RSRM) forward segment propellant grain to control propellant surface burn area. During fabrication, the propellant surface is trimmed prior to the inhibitor application. This produces a potential for small propellant chips to remain undetected on the propellant surface and contaminate the inhibitor during application. The concern was that undetected propellant chips in the inhibitor might provide a fuse path for premature propellant ignition underneath the inhibitor. To evaluate the fuse path potential, testing was performed on inhibitor samples with embedded propellant. The internal motor environment was simulated with a calibrated CO2 laser beam directed onto a sample which was placed in a 4100 kPa (600 psi) nitrogen pressurized bomb (laser bomb). The testing showed definitive results pertaining to fuse path formation. Embedded propellant chips did not autoignite until the receding heat affected inhibitor surface reached, or passed, the propellant chip. Samples with embedded propellant chips in alignment did not propagate ignition from one chip to another with separation distances as small as 0.010 cm(0.004 inc) and some as little as 0.0051 cm (0.002 in). Propellant chips with volumes approximately less than 0.025 cu cm (0.0015 cu in) (which did not propagate ignition) did not increase the inhibitor material decomposition depth more than the resulting void cavity of the burned out propellant chip. In addition, the depth of this void cavity did not increase until it was overtaken by the surrounding material decomposition depth. This was due, in part, to the retention of the protective inhibitor char layer. Samples with embedded propellant strings, whose thicknesses were below 0.023 cm (0.009 in), did not propagate ignition. Propellant string thicknesses above 0.038 cm (0.015 in) did propagate ignition. Test sample char and heat affected layer measurements and observations compared well with those from the Space Shuttle Solid Rocket Motor (SRM) Technical Evaluation Motor no. 9(TEM-9).

  8. Delamination study of chip-to-chip bonding for a LIGA-based safety and arming system

    NASA Astrophysics Data System (ADS)

    Subramanian, Gowrishankar; Deeds, Michael; Cochran, Kevin R.; Raghavan, Raghu; Sandborn, Peter A.

    1999-08-01

    The development of a miniature underwater weapon safety and arming system requires reliable chip-to-chip bonding of die that contain microelectromechanical actuators and sensors fabricated using a LIGA MEMS fabrication process. Chip-to- chip bonding is associated for several different bond materials (indium solder, thermoplastic paste, thermoplastic film and epoxy film), and bonding configurations (with an alloy 42 spacer, silicon to ceramic, and silicon to silicon). Metrology using acoustic micro imaging has been developed to determine the fraction of delamination of samples.

  9. Development of a novel protein chip for the detection of bluetongue virus in China.

    PubMed

    Xu, Q Y; Sun, E C; Feng, Y F; Li, J P; Lv, S; Zhang, Q; Wang, H X; Zhang, J K; Wu, D L

    2016-08-01

    Bluetongue (BT), which is caused by the BT virus (BTV), is an important disease in ruminants that leads to significant economic losses in the husbandry industry. To detect BTV-specific antibodies in serum, a protein chip detection method based on a novel solid supporting material known as polymer-coated initiator-integrated poly (dimethyl siloxane) (iPDMS) was developed. With a threshold of 25% (signal-to-noise percentage), the sensitivity and specificity of the protein chip were 98.6% and 94.8%, respectively. Furthermore, spot serum samples obtained from six provinces of China were tested with the protein chip and a commercially available BTV enzyme-linked immunosorbent assay (ELISA) kit (IDEXX). Of 615 samples, BTV-specific antibodies were detected in 200 (32.52%) by the protein chip and in 176 (28.62%) by the IDEXX BTV ELISA kit. Comparison of the protein chip with the commercial IDEXX BTV ELISA kit yielded the following spot serum detection results: a total coincidence, a negative coincidence and a positive coincidence of 95.12%, 99.28% and 86.5%, respectively. With the protein chip, the BTV-specific serum antibody was detected in samples from all six provinces, and the positive rates ranged from 4.12 to 74.4%. These results indicate that this protein chip detection method based on iPDMS is useful for the serological diagnosis of BTV infection and for epidemiological investigation. Copyright © 2016. Published by Elsevier B.V.

  10. Terahertz microfluidic chips for detection of amino acids in aqueous solutions

    NASA Astrophysics Data System (ADS)

    Su, Bo; Zhang, Cong; Fan, Ning; Zhang, Cunlin

    2016-11-01

    Microfluidic technology can control the fluidic thickness accurately in less than 100 micrometers. So the combination of terahertz (THz) and microfluidic technology becomes one of the most interesting directions towards biological detection. We designed microfluidic chips for terahertz spectroscopy of biological samples in aqueous solutions. Using the terahertz time-domain spectroscopy (THz-TDS) system, we experimentally measured the transmittance of the chips and the THz absorption spectra of L-threonine and L-arginine, respectively. The results indicated the feasibility of performing high sensitivity THz spectroscopy of amino acids solutions. Therefore, the microfluidic chips can realize real-time and label-free measurement for biochemistry samples in THz-TDS system.

  11. Consumption study and identification of methyl salicylate in spicy cassava chips

    NASA Astrophysics Data System (ADS)

    Nirjana, Marlene; Anggadiredja, Kusnandar; Damayanti, Sophi

    2015-09-01

    Spicy cassava chips is a popular snack. However, some news in electronic media reported addition of balsam which is a banned food additives in that product to give extra spicy flavor. This study aimed to determine ITB students' pattern of consumption, health problems caused by spicy chips consumption, and knowledge about illicit use of food additives in that product, and identify the main content of balsam namely methyl salicylate in 10 samples of spicy cassava chips taken from inside and outside about ITB campus. A total of 300 questionnaires distributed to ITB students then data processing was performed. Spicy cassava chips sample macerated in 50 mL of methanol for 24 hours at room temperature, filtered and analyzed using gas chromatography capillary column with OV-1, nitrogen carrier gas and flame ionization detector. Based on questionnaires, 292 (97%) of 300 respondents had consumed spicy chips. A total of 247 (85%) from 292 respondents spicy chips consumed less than 3 times a week. A total of 195 respondents (67%) had experienced health problems after eating spicy chips. There were 137 (47%) of the 292 respondents who knew about the illicit addition of food additives into spicy chips; only 35 respondents (12%) who knew about balsam's addition. There were 126 respondents (43%) who did not pay attention to their health because they will keep eating spicy chips despite the addition of banned food additives. Through the verification of the standard addition method in gas chromatography system with a hydrogen pressure of 1.5 bar, injector temperature 200 °C, detector temperature 230 °C, oven temperature 60 °C for 2 minutes and then increased to 230 °C with rate 6 °C/menit; linearity, limit of detection, limit of quantitation, accuracy, precision, and specificity parameters met the acceptance limits. From 10 spicy cassava chips samples which were analyzed, they did not reveal any content of methyl salicylate. Methyl salicylate contained in the positive control was 1.273 mg/mL.

  12. An analog silicon retina with multichip configuration.

    PubMed

    Kameda, Seiji; Yagi, Tetsuya

    2006-01-01

    The neuromorphic silicon retina is a novel analog very large scale integrated circuit that emulates the structure and the function of the retinal neuronal circuit. We fabricated a neuromorphic silicon retina, in which sample/hold circuits were embedded to generate fluctuation-suppressed outputs in the previous study [1]. The applications of this silicon retina, however, are limited because of a low spatial resolution and computational variability. In this paper, we have fabricated a multichip silicon retina in which the functional network circuits are divided into two chips: the photoreceptor network chip (P chip) and the horizontal cell network chip (H chip). The output images of the P chip are transferred to the H chip with analog voltages through the line-parallel transfer bus. The sample/hold circuits embedded in the P and H chips compensate for the pattern noise generated on the circuits, including the analog communication pathway. Using the multichip silicon retina together with an off-chip differential amplifier, spatial filtering of the image with an odd- and an even-symmetric orientation selective receptive fields was carried out in real time. The analog data transfer method in the present multichip silicon retina is useful to design analog neuromorphic multichip systems that mimic the hierarchical structure of neuronal networks in the visual system.

  13. Trapping ultracold gases near cryogenic materials with rapid reconfigurability

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Naides, Matthew A.; Turner, Richard W.; Lai, Ruby A.

    We demonstrate an atom chip trapping system that allows the placement and high-resolution imaging of ultracold atoms within microns from any ≲100 μm-thin, UHV-compatible material, while also allowing sample exchange with minimal experimental downtime. The sample is not connected to the atom chip, allowing rapid exchange without perturbing the atom chip or laser cooling apparatus. Exchange of the sample and retrapping of atoms has been performed within a week turnaround, limited only by chamber baking. Moreover, the decoupling of sample and atom chip provides the ability to independently tune the sample temperature and its position with respect to the trapped ultracoldmore » gas, which itself may remain in the focus of a high-resolution imaging system. As a first demonstration of this system, we have confined a 700-nK cloud of 8 × 10{sup 4} {sup 87}Rb atoms within 100 μm of a gold-mirrored 100-μm-thick silicon substrate. The substrate was cooled to 35 K without use of a heat shield, while the atom chip, 120 μm away, remained at room temperature. Atoms may be imaged and retrapped every 16 s, allowing rapid data collection.« less

  14. A volumetric meter chip for point-of-care quantitative detection of bovine catalase for food safety control.

    PubMed

    Cui, Xingye; Hu, Jie; Choi, Jane Ru; Huang, Yalin; Wang, Xuemin; Lu, Tian Jian; Xu, Feng

    2016-09-07

    A volumetric meter chip was developed for quantitative point-of-care (POC) analysis of bovine catalase, a bioindicator of bovine mastitis, in milk samples. The meter chip displays multiplexed quantitative results by presenting the distance of ink bar advancement that is detectable by the naked eye. The meter chip comprises a poly(methyl methacrylate) (PMMA) layer, a double-sided adhesive (DSA) layer and a glass slide layer fabricated by the laser-etching method, which is typically simple, rapid (∼3 min per chip), and cost effective (∼$0.2 per chip). Specially designed "U shape" reaction cells are covered by an adhesive tape that serves as an on-off switch, enabling the simple operation of the assay. As a proof of concept, we employed the developed meter chip for the quantification of bovine catalase in raw milk samples to detect catalase concentrations as low as 20 μg/mL. The meter chip has great potential to detect various target analytes for a wide range of POC applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Electrokinetic focusing injection methods on microfluidic devices.

    PubMed

    Fu, Lung-Ming; Yang, Ruey-Jen; Lee, Gwo-Bin

    2003-04-15

    This paper presents an experimental and numerical investigation into electrokinetic focusing injection on microfluidic chips. The valving characteristics on microfluidic devices are controlled through appropriate manipulations of the electric potential strengths during the sample loading and dispensing steps. The present study also addresses the design and testing of various injection systems used to deliver a sample plug. A novel double-cross injection microfluidic chip is fabricated, which employs electrokinetic focusing to deliver sample plugs of variable volume. The proposed design combines several functions of traditional sample plug injection systems on a single microfluidic chip. The injection technique uses an unique sequence of loading steps with different electric potential distributions and magnitudes within the various channels to effectuate a virtual valve.

  16. Color measurement of methylene blue dye/clay mixtures and its application using economical methods

    NASA Astrophysics Data System (ADS)

    Milosevic, Maja; Kaludjerovic, Lazar; Logar, Mihovil

    2016-04-01

    Identifying the clay mineral components of clay materials by staining tests is rapid and simple, but their applicability is restricted because of the mutual interference of the common components of clay materials and difficulties in color determination. The change of color with concentration of the dye is related to the use of colorants as a field test for identifying clay minerals and has been improved over the years to assure the accuracy of the tests (Faust G. T., 1940). The problem of measurement and standardization of color may be solved by combination of colors observed in staining tests with prepared charts of color chips available in the Munsell Book of Color, published by Munsell Color Co. Under a particular set of illumination conditions, a human eye can achieve an approximate match between the color of the dyed clay sample and that of a standard color chip, even though they do have different spectral reflectance characteristics. Experiments were carried out with diffuse reflectance spectroscopy on selected clay samples (three montmorillonite, three kaolinite and one mix-layer clay samples) saturated with different concentration of methylene blue dye solution. Dominant wavelength and purity of the color was obtained on oriented dry samples and calculated by use of the I. C. I. (x, y) - diagram in the region of 400-700 nm (reflectance spectra) without MB and after saturation with different concentrations of MB solutions. Samples were carefully photographed in the natural light environment and processed with user friendly and easily accessible applications (Adobe color CC and ColorHexa encyclopedia) available for android phones or tablets. Obtained colors were compared with Munsell standard color chips, RGB and Hexa color standards. Changes in the color of clay samples in their interaction with different concentration of the applied dye together with application of economical methods can still be used as a rapid fieldwork test. Different types of clay minerals can be distinguished by application of at least three concentrations of the methylene blue dye on the same sample and observing the color change in comparison with standardized color chips that can be easily obtained and free of charge. If the color tests are properly used in conjunction with other more complex analytical procedures, they can be helpful addition in identification of different clay minerals, especially montmorillonite and kaolinite minerals. - Faust G. T., 1940, Staining of clay minerals as a rapid means of identification in natural and beneficiated products, U. S. Bur. Mines, Investigation Report. N0.3522 - Munsell Color, Munsell Book of Color, 1942. Macbeth Division of Kollmorgen Corporation, Maryland, U.S.A. - https://color.adobe.com/create/color-wheel/ - http://www.colorhexa.com/

  17. Tagging of Test Tubes with Electronic p-Chips for Use in Biorepositories.

    PubMed

    Mandecki, Wlodek; Kopacka, Wesley M; Qian, Ziye; Ertwine, Von; Gedzberg, Katie; Gruda, Maryann; Reinhardt, David; Rodriguez, Efrain

    2017-08-01

    A system has been developed to electronically tag and track test tubes used in biorepositories. The system is based on a light-activated microtransponder, also known as a "p-Chip." One of the pressing problems with storing and retrieving biological samples at low temperatures is the difficulty of reliably reading the identification (ID) number that links each storage tube with the database containing sample details. Commonly used barcodes are not always reliable at low temperatures because of poor adhesion of the label to the test tube and problems with reading under conditions of frost and ice accumulation. Traditional radio frequency identification (RFID) tags are not cost effective and are too large for this application. The system described herein consists of the p-Chip, p-Chip-tagged test tubes, two ID readers (for single tubes or for racks of tubes), and software. We also describe a robot that is configured for retrofitting legacy test tubes in biorepositories with p-Chips while maintaining the temperature of the sample below -50°C at all times. The main benefits of the p-Chip over other RFID devices are its small size (600 × 600 × 100 μm) that allows even very small tubes or vials to be tagged, low cost due to the chip's unitary construction, durability, and the ability to read the ID through frost and ice.

  18. Acrylamide in Romanian food using HPLC-UV and a health risk assessment.

    PubMed

    Oroian, Mircea; Amariei, Sonia; Gutt, Gheorghe

    2015-01-01

    The aim of this study was to investigate the level of acrylamide from coffee, potato chips and French fries in Romanian food. According to the European Food Safety Authority, coffee beans, potato chips and French fries have the highest levels of acrylamide. For this survey, 50 samples of coffee beans, 50 samples of potato chips and 25 samples of French fries were purchased from different producers from the Romanian market. Acrylamide levels have been quantified using high-performance liquid chromatography with a diode array detector (HPLC-DAD) method, using water as mobile phase. Health risk assessment was achieved by computing the average daily intake, hazard quotient, cumulative risk, carcinogenic risk and cancer risk. For coffee, potato chips and French fries, acrylamide was not shown to pose a health risk in Romanian food.

  19. Lab-on-a-Chip Application Development-Portable Test System (LOCAD) Phase 2

    NASA Image and Video Library

    2009-03-21

    ISS018-E-041370 (21 March 2009) --- Astronaut Sandra Magnus, STS-119 mission specialist, prepares to work with the Lab-on-a-Chip Application Development-Portable Test System (LOCAD-PTS) experiment in the Destiny laboratory while Space Shuttle Discovery remains docked with the International Space Station. LOCAD-PTS is a handheld device for rapid detection of biological and chemical substances onboard the station.

  20. Pest risk assessment of the importation into the United States of unprocessed Eucalyptus logs and chips from South America

    Treesearch

    John T. Kliejunas; Harold H. Burdsall; Gregg A. DeNitto; Andris Eglitis; Dennis A. Haugen; William E. Wallner

    2001-01-01

    In this report, we assess the unmitigated pest risk potential of importing Eucalyptus logs and chips from South America into the United States. To do this, we estimated the likelihood and consequences of introducing representative insects and pathogens of concern. Nineteen individual pest risk assessments were prepared, eleven dealing with insects and eight with...

  1. Lateral hydrogen microsensors prepared on-chip by local oxidation of platinum-decorated titanium films

    NASA Astrophysics Data System (ADS)

    Herbertz, S.; Welk, D.; Heinzel, T.

    2018-05-01

    Titanium microstripes on silicon dioxide substrates are oxidized locally by applying voltages on-chip to lateral electrodes under ambient conditions. This technique enables profound modifications of the electronic circuit. As an example, we transform Ti films decorated by a sub-monolayer of platinum into hydrogen gas microsensors in an otherwise completed device by a silicon-MOS compatible process.

  2. Programmable lab-on-a-chip system for single cell analysis

    NASA Astrophysics Data System (ADS)

    Thalhammer, S.

    2009-05-01

    The collection, selection, amplification and detection of minimum genetic samples became a part of everyday life in medical and biological laboratories, to analyze DNA-fragments of pathogens, patient samples and traces on crime scenes. About a decade ago, a handful of researchers began discussing an intriguing idea. Could the equipment needed for everyday chemistry and biology procedures be shrunk to fit on a chip in the size of a fingernail? Miniature devices for, say, analysing DNA and proteins should be faster and cheaper than conventional versions. Lab-on-a-chip is an advanced technology that integrates a microfluidic system on a microscale chip device. The "laboratory" is created by means of channels, mixers, reservoirs, diffusion chambers, integrated electrodes, pumps, valves and more. With lab-ona- chip technology, complete laboratories on a square centimetre can be created. Here, a multifunctional programmable Lab-on-a-Chip driven by nanofluidics and controlled by surface acoustic waves (SAW) is presented. This system combines serial DNA-isolation-, amplification- and array-detection-process on a modified glass-platform. The fluid actuation is controlled via SAW by interdigital transducers implemented in the chemical modified chip surface. The chemical surface modification allows fluid handling in the sub-microliter range. Minute amount of sample material is extracted by laser-based microdissection out of e.g. histological sections at the single cell level. A few picogram of genetic material are isolated and transferred via a low-pressure transfer system (SPATS) onto the chip. Subsequently the genetic material inside single droplets, which behave like "virtual" beaker, is transported to the reaction and analysis centers on the chip surface via surface acoustic waves, mainly known as noise dumping filters in mobile phones. At these "biological reactors" the genetic material is processed, e.g. amplified via polymerase chain reaction methods, and genetically characterized.

  3. The correlation of methylation levels measured using Illumina 450K and EPIC BeadChips in blood samples

    PubMed Central

    Logue, Mark W; Smith, Alicia K; Wolf, Erika J; Maniates, Hannah; Stone, Annjanette; Schichman, Steven A; McGlinchey, Regina E; Milberg, William; Miller, Mark W

    2017-01-01

    Aim: We examined concordance of methylation levels across the Illumina Infinium HumanMethylation450 BeadChip and the Infinium MethylationEPIC BeadChip. Methods: We computed the correlation for 145 whole blood DNA samples at each of the 422,524 CpG sites measured by both chips. Results: The correlation at some sites was high (up to r = 0.95), but many sites had low correlation (55% had r < 0.20). The low correspondence between 450K and EPIC measured methylation values at many loci was largely due to the low variability in methylation values for the majority of the CpG sites in blood. Conclusion: Filtering out probes based on the observed correlation or low variability may increase reproducibility of BeadChip-based epidemiological studies. PMID:28809127

  4. Application of an improved magnetic immunosorbent in an Ephesia chip designed for circulating tumor cell capture.

    PubMed

    Svobodova, Zuzana; Kucerova, Jana; Autebert, Julien; Horak, Daniel; Bruckova, Lenka; Viovy, Jean-Louis; Bilkova, Zuzana

    2014-02-01

    In this study, we describe a particular step in developing a microfluidic device for capture and detection of circulating tumor cells-specifically the preparation of an immunosorbent for implementation into the separation chip. We highlight some of the most important specifics connected with superparamegnetic microspheres for microfluidic purposes. Factors such as nonspecific adsorption on microfluidic channels, interactions with model cell lines, and tendency to aggregation were investigated. Poly(glycidyl methacrylate) microspheres with carboxyl groups were employed for this purpose. To address the aforementioned challenges, the microspheres were coated with hydrazide-PEG-hydrazide, and subsequently anti-epithelial cell adhesion molecule (EpCAM) antibody was immobilized. The prepared anti-EpCAM immunosorbent was pretested using model cell lines with differing EpCAM density (MCF7, SKBR3, A549, and Raji) in a batchwise arrangement. Finally, the entire system was implemented and studied in an Ephesia chip and an evaluation was performed by the MCF7 cell line. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. A 1 GHz sample rate, 256-channel, 1-bit quantization, CMOS, digital correlator chip

    NASA Technical Reports Server (NTRS)

    Timoc, C.; Tran, T.; Wongso, J.

    1992-01-01

    This paper describes the development of a digital correlator chip with the following features: 1 Giga-sample/second; 256 channels; 1-bit quantization; 32-bit counters providing up to 4 seconds integration time at 1 GHz; and very low power dissipation per channel. The improvements in the performance-to-cost ratio of the digital correlator chip are achieved with a combination of systolic architecture, novel pipelined differential logic circuits, and standard 1.0 micron CMOS process.

  6. A microfluidic thermometer: Precise temperature measurements in microliter- and nanoliter-scale volumes

    PubMed Central

    McKenzie, Brittney A.

    2017-01-01

    Measuring the temperature of a sample is a fundamental need in many biological and chemical processes. When the volume of the sample is on the microliter or nanoliter scale (e.g., cells, microorganisms, precious samples, or samples in microfluidic devices), accurate measurement of the sample temperature becomes challenging. In this work, we demonstrate a technique for accurately determining the temperature of microliter volumes using a simple 3D-printed microfluidic chip. We accomplish this by first filling “microfluidic thermometer” channels on the chip with substances with precisely known freezing/melting points. We then use a thermoelectric cooler to create a stable and linear temperature gradient along these channels within a measurement region on the chip. A custom software tool (available as online Supporting Information) is then used to find the locations of solid-liquid interfaces in the thermometer channels; these locations have known temperatures equal to the freezing/melting points of the substances in the channels. The software then uses the locations of these interfaces to calculate the temperature at any desired point within the measurement region. Using this approach, the temperature of any microliter-scale on-chip sample can be measured with an uncertainty of about a quarter of a degree Celsius. As a proof-of-concept, we use this technique to measure the unknown freezing point of a 50 microliter volume of solution and demonstrate its feasibility on a 400 nanoliter sample. Additionally, this technique can be used to measure the temperature of any on-chip sample, not just near-zero-Celsius freezing points. We demonstrate this by using an oil that solidifies near room temperature (coconut oil) in a microfluidic thermometer to measure on-chip temperatures well above zero Celsius. By providing a low-cost and simple way to accurately measure temperatures in small volumes, this technique should find applications in both research and educational laboratories. PMID:29284028

  7. Differential alternating current chip calorimeter for in situ investigation of vapor-deposited thin films

    NASA Astrophysics Data System (ADS)

    Ahrenberg, M.; Shoifet, E.; Whitaker, K. R.; Huth, H.; Ediger, M. D.; Schick, C.

    2012-03-01

    Physical vapor deposition can be used to produce thin films with interesting material properties including extraordinarily stable organic glasses. We describe an ac chip calorimeter for in situ heat capacity measurements of as-deposited nanometer thin films of organic glass formers. The calorimetric system is based on a differential ac chip calorimeter which is placed in the vacuum chamber for physical vapor deposition. The sample is directly deposited onto one calorimetric chip sensor while the other sensor is protected against deposition. The device and the temperature calibration procedure are described. The latter makes use of the phase transitions of cyclopentane and the frequency dependence of the dynamic glass transition of toluene and ethylbenzene. Sample thickness determination is based on a finite element modeling of the sensor sample arrangement. In the modeling, a layer of toluene was added to the sample sensor and its thickness was varied in an iterative way until the model fit the experimental data.

  8. Differential AC chip calorimeter for in situ investigation of vapor deposited thin films

    NASA Astrophysics Data System (ADS)

    Ahrenberg, Mathias; Schick, Christoph; Huth, Heiko; Schoifet, Evgeni; Ediger, Mark; Whitaker, Katie

    2012-02-01

    Physical vapor deposition (PVD) can be used to produce thin films with particular material properties like extraordinarily stable glasses of organic molecules. We describe an AC chip calorimeter for in-situ heat capacity measurements of as-deposited nanometer thin films of organic glass formers. The calorimetric system is based on a differential AC chip calorimeter which is placed in the vacuum chamber for physical vapor deposition. The sample is directly deposited onto one calorimetric chip sensor while the other sensor is protected against deposition. The device and the temperature calibration procedure are described. The latter makes use of the phase transitions of cyclopentane and the frequency dependence of the dynamic glass transition of toluene and ethylbenzene. Sample thickness determination is based on a finite element modeling (FEM) of the sensor sample arrangement. A layer of toluene was added to the sample sensor and its thickness was varied in an iterative way until the model fits the experimental data.

  9. On-chip purification and detection of hepatitis C virus RNA from human plasma.

    PubMed

    Vaghi, V; Potrich, C; Pasquardini, L; Lunelli, L; Vanzetti, L; Ebranati, E; Lai, A; Zehender, G; Mombello, D; Cocuzza, M; Pirri, C F; Pederzolli, C

    2016-01-01

    Hepatitis C virus (HCV) is one of the main causes of chronic liver disease worldwide. The diagnosis and monitoring of HCV infection is a crucial need in the clinical management. The conventional diagnostic technologies are challenged when trying to address molecular diagnostics, especially because they require a complex and time-consuming sample preparation phase. Here, a new concept based on surface functionalization was applied to viral RNA purification: first of all polydimethylsiloxane (PDMS) flat surfaces were modified to hold RNA adsorption. After a careful chemical and morphological analysis of the modified surfaces, the functionalization protocols giving the best RNA adsorbing surfaces were applied to PDMS microdevices. The functionalized microdevices were then used for RNA purification from HCV infected human plasma samples. RNA purification and RT were successfully performed in the same microdevice chamber, saving time of analysis, reagents, and labor. The PCR protocol for HCV cDNA amplification was also implemented in the microdevice, demonstrating that the entire process of HCV analysis, from plasma to molecular readout, could be performed on-chip. Not only HCV but also other microdevice-based viral RNA detection could therefore result in a successful Point-of-Care (POC) diagnostics for resource-limited settings. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Label free detection of phospholipids by infrared absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Ahmed, Tahsin; Foster, Erick; Vigil, Genevieve; Khan, Aamir A.; Bohn, Paul; Howard, Scott S.

    2014-08-01

    We present our study on compact, label-free dissolved lipid sensing by combining capillary electrophoresis separation in a PDMS microfluidic chip online with mid-infrared (MIR) absorption spectroscopy for biomarker detection. On-chip capillary electrophoresis is used to separate the biomarkers without introducing any extrinsic contrast agent, which reduces both cost and complexity. The label free biomarker detection could be done by interrogating separated biomarkers in the channel by MIR absorption spectroscopy. Phospholipids biomarkers of degenerative neurological, kidney, and bone diseases are detectable using this label free technique. These phospholipids exhibit strong absorption resonances in the MIR and are present in biofluids including urine, blood plasma, and cerebrospinal fluid. MIR spectroscopy of a 12-carbon chain phosphatidic acid (PA) (1,2-dilauroyl-snglycero- 3-phosphate (sodium salt)) dissolved in N-methylformamide, exhibits a strong amide peak near wavenumber 1660 cm-1 (wavelength 6 μm), arising from the phosphate headgroup vibrations within a low-loss window of the solvent. PA has a similar structure to many important phospholipids molecules like phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), and phosphatidylserine (PS), making it an ideal molecule for initial proof-of-concept studies. This newly proposed detection technique can lead us to minimal sample preparation and is capable of identifying several biomarkers from the same sample simultaneously.

  11. Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling

    NASA Astrophysics Data System (ADS)

    Shang, Yuqin; Zeng, Yun; Zeng, Yong

    2016-02-01

    Protein glycosylation is one of the key processes that play essential roles in biological functions and dysfunctions. However, progress in glycomics has considerably lagged behind genomics and proteomics, due in part to the enormous challenges in analysis of glycans. Here we present a new integrated and automated microfluidic lectin barcode platform to substantially improve the performance of lectin array for focused glycomic profiling. The chip design and flow control were optimized to promote the lectin-glycan binding kinetics and speed of lectin microarray. Moreover, we established an on-chip lectin assay which employs a very simple blocking method to effectively suppress the undesired background due to lectin binding of antibodies. Using this technology, we demonstrated focused differential profiling of tissue-specific glycosylation changes of a biomarker, CA125 protein purified from ovarian cancer cell line and different tissues from ovarian cancer patients in a fast, reproducible, and high-throughput fashion. Highly sensitive CA125 detection was also demonstrated with a detection limit much lower than the clinical cutoff value for cancer diagnosis. This microfluidic platform holds the potential to integrate with sample preparation functions to construct a fully integrated “sample-to-answer” microsystem for focused differential glycomic analysis. Thus, our technology should present a powerful tool in support of rapid advance in glycobiology and glyco-biomarker development.

  12. Double-helix micro-channels on microfluidic chips for enhanced continuous on-chip derivatization followed by electrophoretic separation.

    PubMed

    Peng, Xianglu; Zhao, Lei; Guo, Jinxiu; Yang, Shenghong; Ding, Hui; Wang, Xiayan; Pu, Qiaosheng

    2015-10-15

    Micro-channels that contain a special inner structure are critical for efficient mixing and chemical reactions. In this paper, we described the facile fabrication of an integrated microchip with double-helix type micro-channels to improve mixing efficiency and to facilitate multi-step derivatization reactions prior to electrophoretic separation. With a prepared microchip, reagents, samples and reaction products could be driven through micro-channels by siphon, and no other pumping device was necessary. To test its performance, reductive amination of aldehydes with 8-aminonaphthalene-1,3,6-trisulfonate acid disodium (ANTS) was attempted via microchip electrophoresis with laser induced fluorescence (LIF). The effect of the geometry of the reaction micro-channel on the reaction's efficiency was evaluated. Under the selected conditions, successful derivatization of five aldehydes was realized for highly reproducible analysis. The relative standard deviations of the peak areas for 30 consecutive injections were in the range of 0.28-1.61%. The method was applied for the determination of aldehydes in real samples with standard addition recoveries of 87.8-102.8%. Good tolerance of organic solvents was achieved, and the proposed method can potentially be employed for rapid screening of excessively added aldehyde food flavoring. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling

    PubMed Central

    Shang, Yuqin; Zeng, Yun; Zeng, Yong

    2016-01-01

    Protein glycosylation is one of the key processes that play essential roles in biological functions and dysfunctions. However, progress in glycomics has considerably lagged behind genomics and proteomics, due in part to the enormous challenges in analysis of glycans. Here we present a new integrated and automated microfluidic lectin barcode platform to substantially improve the performance of lectin array for focused glycomic profiling. The chip design and flow control were optimized to promote the lectin-glycan binding kinetics and speed of lectin microarray. Moreover, we established an on-chip lectin assay which employs a very simple blocking method to effectively suppress the undesired background due to lectin binding of antibodies. Using this technology, we demonstrated focused differential profiling of tissue-specific glycosylation changes of a biomarker, CA125 protein purified from ovarian cancer cell line and different tissues from ovarian cancer patients in a fast, reproducible, and high-throughput fashion. Highly sensitive CA125 detection was also demonstrated with a detection limit much lower than the clinical cutoff value for cancer diagnosis. This microfluidic platform holds the potential to integrate with sample preparation functions to construct a fully integrated “sample-to-answer” microsystem for focused differential glycomic analysis. Thus, our technology should present a powerful tool in support of rapid advance in glycobiology and glyco-biomarker development. PMID:26831207

  14. In vitro RNA release from a human rhinovirus monitored by means of a molecular beacon and chip electrophoresis.

    PubMed

    Weiss, Victor U; Bliem, Christina; Gösler, Irene; Fedosyuk, Sofiya; Kratzmeier, Martin; Blaas, Dieter; Allmaier, Günter

    2016-06-01

    Liquid-phase electrophoresis either in the classical capillary format or miniaturized (chip CE) is a valuable tool for quality control of virus preparations and for targeting questions related to conformational changes of viruses during infection. We present an in vitro assay to follow the release of the RNA genome from a human rhinovirus (common cold virus) by using a molecular beacon (MB) and chip CE. The MB, a probe that becomes fluorescent upon hybridization to a complementary sequence, was designed to bind close to the 3' end of the viral genome. Addition of Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), a well-known additive for reduction of bleaching and blinking of fluorophores in fluorescence microscopy, to the background electrolyte increased the sensitivity of our chip CE set-up. Hence, a fast, sensitive and straightforward method for the detection of viral RNA is introduced. Additionally, challenges of our assay will be discussed. In particular, we found that (i) desalting of virus preparations prior to analysis increased the recorded signal and (ii) the MB-RNA complex signal decreased with the time of virus storage at -70 °C. This suggests that 3'-proximal sequences of the viral RNA, if not the whole genome, underwent degradation during storage and/or freezing and thawing. In summary, we demonstrate, for two independent virus batches, that chip electrophoresis can be used to monitor MB hybridization to RNA released upon incubation of the native virus at 56 °C. Graphical Abstract Schematic of the study strategy: RNA released from HRV-A2 is detected by chip electrophoresis through the increase in fluorescence after genom complexation to a cognate molecular beacon.

  15. New immobilisation protocol for the template used in solid-phase synthesis of MIP nanoparticles

    NASA Astrophysics Data System (ADS)

    Chen, Lu; Muhammad, Turghun; Yakup, Burabiye; Piletsky, Sergey A.

    2017-06-01

    As a novel imprinting method, solid-phase synthesis has proven to be a promising approach to prepare polymer nanoparticles with specific recognition sites for a template molecule. In this method, imprinted polymer nanoparticles were synthesized using template immobilized on a solid support. Herein, preparation of immobilized templates on quartz chips through homogeneous route was reported as an efficient alternative strategy to heterogeneous one. The template molecule indole-3-butyric acid (IBA) was reacted with 3-aminopropyltriethoxysilane (APTES) to produce silylated template (IBA-APTES), and it was characterized by IR, 1H NMR and GC-MS. Then, the silylated template molecule was grafted onto the activated surfaces of quartz chip to prepare immobilized template (SiO2@IBA-APTES). The immobilization was confirmed by contact angle, XPS, UV and fluorescence measurement. Immobilization protocol has shown good reproducibility and stability of the immobilized template. MIP nanoparticles were prepared with high selectivity toward the molecule immobilized onto the solid surface. This provides a new approach for the development of molecularly imprinted nanoparticles.

  16. 3D imaging of optically cleared tissue using a simplified CLARITY method and on-chip microscopy

    PubMed Central

    Zhang, Yibo; Shin, Yoonjung; Sung, Kevin; Yang, Sam; Chen, Harrison; Wang, Hongda; Teng, Da; Rivenson, Yair; Kulkarni, Rajan P.; Ozcan, Aydogan

    2017-01-01

    High-throughput sectioning and optical imaging of tissue samples using traditional immunohistochemical techniques can be costly and inaccessible in resource-limited areas. We demonstrate three-dimensional (3D) imaging and phenotyping in optically transparent tissue using lens-free holographic on-chip microscopy as a low-cost, simple, and high-throughput alternative to conventional approaches. The tissue sample is passively cleared using a simplified CLARITY method and stained using 3,3′-diaminobenzidine to target cells of interest, enabling bright-field optical imaging and 3D sectioning of thick samples. The lens-free computational microscope uses pixel super-resolution and multi-height phase recovery algorithms to digitally refocus throughout the cleared tissue and obtain a 3D stack of complex-valued images of the sample, containing both phase and amplitude information. We optimized the tissue-clearing and imaging system by finding the optimal illumination wavelength, tissue thickness, sample preparation parameters, and the number of heights of the lens-free image acquisition and implemented a sparsity-based denoising algorithm to maximize the imaging volume and minimize the amount of the acquired data while also preserving the contrast-to-noise ratio of the reconstructed images. As a proof of concept, we achieved 3D imaging of neurons in a 200-μm-thick cleared mouse brain tissue over a wide field of view of 20.5 mm2. The lens-free microscope also achieved more than an order-of-magnitude reduction in raw data compared to a conventional scanning optical microscope imaging the same sample volume. Being low cost, simple, high-throughput, and data-efficient, we believe that this CLARITY-enabled computational tissue imaging technique could find numerous applications in biomedical diagnosis and research in low-resource settings. PMID:28819645

  17. Miniaturized Sample Preparation and Rapid Detection of Arsenite in Contaminated Soil Using a Smartphone.

    PubMed

    Siddiqui, Mohd Farhan; Kim, Soocheol; Jeon, Hyoil; Kim, Taeho; Joo, Chulmin; Park, Seungkyung

    2018-03-04

    Conventional methods for analyzing heavy metal contamination in soil and water generally require laboratory equipped instruments, complex procedures, skilled personnel and a significant amount of time. With the advancement in computing and multitasking performances, smartphone-based sensors potentially allow the transition of the laboratory-based analytical processes to field applicable, simple methods. In the present work, we demonstrate the novel miniaturized setup for simultaneous sample preparation and smartphone-based optical sensing of arsenic As(III) in the contaminated soil. Colorimetric detection protocol utilizing aptamers, gold nanoparticles and NaCl have been optimized and tested on the PDMS-chip to obtain the high sensitivity with the limit of detection of 0.71 ppm (in the sample) and a correlation coefficient of 0.98. The performance of the device is further demonstrated through the comparative analysis of arsenic-spiked soil samples with standard laboratory method, and a good agreement with a correlation coefficient of 0.9917 and the average difference of 0.37 ppm, are experimentally achieved. With the android application on the device to run the experiment, the whole process from sample preparation to detection is completed within 3 hours without the necessity of skilled personnel. The approximate cost of setup is estimated around 1 USD, weight 55 g. Therefore, the presented method offers the simple, rapid, portable and cost-effective means for onsite sensing of arsenic in soil. Combined with the geometric information inside the smartphones, the system will allow the monitoring of the contamination status of soils in a nation-wide manner.

  18. Specific capture, recovery and culture of cancer cells using oriented antibody-modified polystyrene chips coated with agarose film.

    PubMed

    Jeong, Jiyun; Lee, Yeolin; Yoo, Yeongeun; Lee, Myung Kyu

    2018-02-01

    Agarose gel can be used for three dimensional (3D) cell culture because it prevents cell attachment. The dried agarose film coated on a culture plate also protected cell attachment and allowed 3D growth of cancer cells. We developed an efficient method for agarose film coating on an oxygen-plasma treated micropost polystyrene chip prepared by an injection molding process. The agarose film was modified to maleimide or Ni-NTA groups for covalent or cleavable attachment of photoactivatable Fc-specific antibody binding proteins (PFcBPs) via their N-terminal cysteine residues or 6xHis tag, respectively. The antibodies photocrosslinked onto the PFcBP-modified chips specifically captured the target cells without nonspecific binding, and the captured cells grew 3D modes on the chips. The captured cells on the cleavable antibody-modified chips were easily recovered by treatment of commercial trypsin-EDTA solution. Under fluidic conditions using an antibody-modified micropost chip, the cells were mainly captured on the micropost walls of the chip rather than on the bottom of it. The presented method will also be applicable for immobilization of oriented antibodies on various microfluidic chips with different structures. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Floating Chip Mounting System Driven by Repulsive Force of Permanent Magnets for Multiple On-Site SPR Immunoassay Measurements

    PubMed Central

    Horiuchi, Tsutomu; Tobita, Tatsuya; Miura, Toru; Iwasaki, Yuzuru; Seyama, Michiko; Inoue, Suzuyo; Takahashi, Jun-ichi; Haga, Tsuneyuki; Tamechika, Emi

    2012-01-01

    We have developed a measurement chip installation/removal mechanism for a surface plasmon resonance (SPR) immunoassay analysis instrument designed for frequent testing, which requires a rapid and easy technique for changing chips. The key components of the mechanism are refractive index matching gel coated on the rear of the SPR chip and a float that presses the chip down. The refractive index matching gel made it possible to optically couple the chip and the prism of the SPR instrument easily via elastic deformation with no air bubbles. The float has an autonomous attitude control function that keeps the chip parallel in relation to the SPR instrument by employing the repulsive force of permanent magnets between the float and a float guide located in the SPR instrument. This function is realized by balancing the upward elastic force of the gel and the downward force of the float, which experiences a leveling force from the float guide. This system makes it possible to start an SPR measurement immediately after chip installation and to remove the chip immediately after the measurement with a simple and easy method that does not require any fine adjustment. Our sensor chip, which we installed using this mounting system, successfully performed an immunoassay measurement on a model antigen (spiked human-IgG) in a model real sample (non-homogenized milk) that included many kinds of interfering foreign substances without any sample pre-treatment. The ease of the chip installation/removal operation and simple measurement procedure are suitable for frequent on-site agricultural, environmental and medical testing. PMID:23202030

  20. X-ray transparent microfluidic chip for mesophase-based crystallization of membrane proteins and on-chip structure determination

    DOE PAGES

    Khvostichenko, Daria S.; Schieferstein, Jeremy M.; Pawate, Ashtamurthy S.; ...

    2014-08-21

    Crystallization from lipidic mesophase matrices is a promising route to diffraction-quality crystals and structures of membrane proteins. The microfluidic approach reported here eliminates two bottlenecks of the standard mesophase-based crystallization protocols: (i) manual preparation of viscous mesophases and (ii) manual harvesting of often small and fragile protein crystals. In the approach reported here, protein-loaded mesophases are formulated in an X-ray transparent microfluidic chip using only 60 nL of the protein solution per crystallization trial. The X-ray transparency of the chip enables diffraction data collection from multiple crystals residing in microfluidic wells, eliminating the normally required manual harvesting and mounting ofmore » individual crystals. In addition, we validated our approach by on-chip crystallization of photosynthetic reaction center, a membrane protein from Rhodobacter sphaeroides, followed by solving its structure to a resolution of 2.5 Å using X-ray diffraction data collected on-chip under ambient conditions. A moderate conformational change in hydrophilic chains of the protein was observed when comparing the on-chip, room temperature structure with known structures for which data were acquired under cryogenic conditions.« less

  1. X-ray Transparent Microfluidic Chip for Mesophase-Based Crystallization of Membrane Proteins and On-Chip Structure Determination

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Khvostichenko, Daria S.; Schieferstein, Jeremy M.; Pawate, Ashtamurthy S.

    2014-10-01

    Crystallization from lipidic mesophase matrices is a promising route to diffraction-quality crystals and structures of membrane proteins. The microfluidic approach reported here eliminates two bottlenecks of the standard mesophase-based crystallization protocols: (i) manual preparation of viscous mesophases and (ii) manual harvesting of often small and fragile protein crystals. In the approach reported here, protein-loaded mesophases are formulated in an X-ray transparent microfluidic chip using only 60 nL of the protein solution per crystallization trial. The X-ray transparency of the chip enables diffraction data collection from multiple crystals residing in microfluidic wells, eliminating the normally required manual harvesting and mounting ofmore » individual crystals. We validated our approach by on-chip crystallization of photosynthetic reaction center, a membrane protein from Rhodobacter sphaeroides, followed by solving its structure to a resolution of 2.5 Å using X-ray diffraction data collected on-chip under ambient conditions. A moderate conformational change in hydrophilic chains of the protein was observed when comparing the on-chip, room temperature structure with known structures for which data were acquired under cryogenic conditions.« less

  2. A reliability evaluation methodology for memory chips for space applications when sample size is small

    NASA Technical Reports Server (NTRS)

    Chen, Y.; Nguyen, D.; Guertin, S.; Berstein, J.; White, M.; Menke, R.; Kayali, S.

    2003-01-01

    This paper presents a reliability evaluation methodology to obtain the statistical reliability information of memory chips for space applications when the test sample size needs to be kept small because of the high cost of the radiation hardness memories.

  3. Rapid, automated, parallel quantitative immunoassays using highly integrated microfluidics and AlphaLISA

    PubMed Central

    Tak For Yu, Zeta; Guan, Huijiao; Ki Cheung, Mei; McHugh, Walker M.; Cornell, Timothy T.; Shanley, Thomas P.; Kurabayashi, Katsuo; Fu, Jianping

    2015-01-01

    Immunoassays represent one of the most popular analytical methods for detection and quantification of biomolecules. However, conventional immunoassays such as ELISA and flow cytometry, even though providing high sensitivity and specificity and multiplexing capability, can be labor-intensive and prone to human error, making them unsuitable for standardized clinical diagnoses. Using a commercialized no-wash, homogeneous immunoassay technology (‘AlphaLISA’) in conjunction with integrated microfluidics, herein we developed a microfluidic immunoassay chip capable of rapid, automated, parallel immunoassays of microliter quantities of samples. Operation of the microfluidic immunoassay chip entailed rapid mixing and conjugation of AlphaLISA components with target analytes before quantitative imaging for analyte detections in up to eight samples simultaneously. Aspects such as fluid handling and operation, surface passivation, imaging uniformity, and detection sensitivity of the microfluidic immunoassay chip using AlphaLISA were investigated. The microfluidic immunoassay chip could detect one target analyte simultaneously for up to eight samples in 45 min with a limit of detection down to 10 pg mL−1. The microfluidic immunoassay chip was further utilized for functional immunophenotyping to examine cytokine secretion from human immune cells stimulated ex vivo. Together, the microfluidic immunoassay chip provides a promising high-throughput, high-content platform for rapid, automated, parallel quantitative immunosensing applications. PMID:26074253

  4. Development of gold nanoparticle-aptamer-based LSPR sensing chips for the rapid detection of Salmonella typhimurium in pork meat.

    PubMed

    Oh, Seo Yeong; Heo, Nam Su; Shukla, Shruti; Cho, Hye-Jin; Vilian, A T Ezhil; Kim, Jinwoon; Lee, Sang Yup; Han, Young-Kyu; Yoo, Seung Min; Huh, Yun Suk

    2017-08-31

    A non-labeled, portable plasmonic biosensor-based device was developed to enable the ultra-sensitive and selective detection of Salmonella typhimurium in pork meat samples. Specifically, a plasmonic sensor, using the self-assembly of gold nanoparticles (AuNPs) to achieve a regulated diameter of 20 nm for the AuNP monolayers, was used to conduct high-density deposition on a transparent substrate, which produced longitudinal wavelength extinction shifts via a localized surface plasmon resonance (LSPR) signal. The developed aptamers conjugated to the LSPR sensing chips revealed an ultra-sensitive upper limit of detection (LOD) of approximately 10 4 cfu/mL for S. typhimurium in pure culture under the optimal assay conditions, with a total analysis time of 30-35 min. When the LSPR sensing chips were applied on artificially contaminated pork meat samples, S. typhimurium in the spiked pork meat samples was also detected at an LOD of 1.0 × 10 4 cfu/mL. The developed method could detect S. typhimurium in spiked pork meat samples without a pre-enrichment step. Additionally, the LSPR sensing chips developed against S. typhimurium were not susceptible to any effect of the food matrix or background contaminant microflora. These findings confirmed that the developed gold nanoparticle-aptamer-based LSPR sensing chips could facilitate sensitive detection of S. typhimurium in food samples.

  5. Next Generation Programmable Bio-Nano-Chip System for On-Site Detection in Oral Fluids.

    PubMed

    Christodoulides, Nicolaos; De La Garza, Richard; Simmons, Glennon W; McRae, Michael P; Wong, Jorge; Newton, Thomas F; Kosten, Thomas R; Haque, Ahmed; McDevitt, John T

    2015-11-23

    Current on-site drug of abuse detection methods involve invasive sampling of blood and urine specimens, or collection of oral fluid, followed by qualitative screening tests using immunochromatographic cartridges. Test confirmation and quantitative assessment of a presumptive positive are then provided by remote laboratories, an inefficient and costly process decoupled from the initial sampling. Recently, a new noninvasive oral fluid sampling approach that is integrated with the chip-based Programmable Bio-Nano-Chip (p-BNC) platform has been developed for the rapid (~ 10 minutes), sensitive detection (~ ng/ml) and quantitation of 12 drugs of abuse. Furthermore, the system can provide the time-course of select drug and metabolite profiles in oral fluids. For cocaine, we observed three slope components were correlated with cocaine-induced impairment using this chip-based p-BNC detection modality. Thus, this p-BNC has significant potential for roadside drug testing by law enforcement officers. Initial work reported on chip-based drug detection was completed using 'macro' or "chip in the lab" prototypes, that included metal encased "flow cells", external peristaltic pumps and a bench-top analyzer system instrumentation. We now describe the next generation miniaturized analyzer instrumentation along with customized disposables and sampling devices. These tools will offer real-time oral fluid drug monitoring capabilities, to be used for roadside drug testing as well as testing in clinical settings as a non-invasive, quantitative, accurate and sensitive tool to verify patient adherence to treatment.

  6. Improvement and Analysis of the Radiation Response of RADFET Dosimeters

    DTIC Science & Technology

    1992-06-15

    TLD ), silicon p-i-n diode responses and silicon calorimetry (AWE Dosimetry Service). Intensive preparations were made by REM and the experiments were...SUB-GROUP dose: RADFET : tactical dosimetry silicon : metal-oxide- 0705 emiconductor (MOS) field effect transistor (FET) : silicon Idioxide space...1.1 Principle of a dosimetry system, based on the RADFET (radiation-sensitive field-effect transistor) (a) microscopic cross-section of chip (b) chip

  7. Self-priming compartmentalization digital LAMP for point-of-care.

    PubMed

    Zhu, Qiangyuan; Gao, Yibo; Yu, Bingwen; Ren, Hao; Qiu, Lin; Han, Sihai; Jin, Wei; Jin, Qinhan; Mu, Ying

    2012-11-21

    Digital nucleic acid amplification provides unprecedented opportunities for absolute nucleic acid quantification by counting of single molecules. This technique is useful for molecular genetic analysis in cancer, stem cell, bacterial, non-invasive prenatal diagnosis in which many biologists are interested. This paper describes a self-priming compartmentalization (SPC) microfluidic chip platform for performing digital loop-mediated amplification (LAMP). The energy for the pumping is pre-stored in the degassed bulk PDMS by exploiting the high gas solubility of PDMS; therefore, no additional structures other than channels and reservoirs are required. The sample and oil are sequentially sucked into the channels, and the pressure difference of gas dissolved in PDMS allows sample self-compartmentalization without the need for further chip manipulation such as with pneumatic microvalves and control systems, and so on. The SPC digital LAMP chip can be used like a 384-well plate, so, the world-to-chip fluidic interconnections are avoided. The microfluidic chip contains 4 separate panels, each panel contains 1200 independent 6 nL chambers and can be used to detect 4 samples simultaneously. Digital LAMP on the microfluidic chip was tested quantitatively by using β-actin DNA from humans. The self-priming compartmentalization behavior is roughly predictable using a two-dimensional model. The uniformity of compartmentalization was analyzed by fluorescent intensity and fraction of volume. The results showed that the feasibility and flexibility of the microfluidic chip platform for amplifying single nucleic acid molecules in different chambers made by diluting and distributing sample solutions. The SPC chip has the potential to meet the requirements of a general laboratory: power-free, valve-free, operating at isothermal temperature, inexpensive, sensitive, economizing labour time and reagents. The disposable analytical devices with appropriate air-tight packaging should be useful for point-of-care, and enabling it to become one of the common tools for biology research, especially, in point-of-care testing.

  8. 42 CFR 431.954 - Basis and scope.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Improper Payments in Medicaid and CHIP § 431.954 Basis and scope. (a) Basis. The statutory bases for this... or child health assistance under CHIP; and State-specific sample sizes for application of the PERM... improper payments in the fee-for-service (FFS) and managed care components of the Medicaid and CHIP...

  9. 42 CFR 431.954 - Basis and scope.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Improper Payments in Medicaid and CHIP § 431.954 Basis and scope. (a) Basis. The statutory bases for this... or child health assistance under CHIP; and State-specific sample sizes for application of the PERM... improper payments in the fee-for-service (FFS) and managed care components of the Medicaid and CHIP...

  10. 42 CFR 431.954 - Basis and scope.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Improper Payments in Medicaid and CHIP § 431.954 Basis and scope. (a) Basis. The statutory bases for this... or child health assistance under CHIP; and State-specific sample sizes for application of the PERM... improper payments in the fee-for-service (FFS) and managed care components of the Medicaid and CHIP...

  11. 42 CFR 431.954 - Basis and scope.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Improper Payments in Medicaid and CHIP § 431.954 Basis and scope. (a) Basis. The statutory bases for this... or child health assistance under CHIP; and State-specific sample sizes for application of the PERM... improper payments in the fee-for-service (FFS) and managed care components of the Medicaid and CHIP...

  12. Microfluidic LC Device with Orthogonal Sample Extraction for On-Chip MALDI-MS Detection

    PubMed Central

    Lazar, Iulia M.; Kabulski, Jarod L.

    2013-01-01

    A microfluidic device that enables on-chip matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS) detection for liquid chromatography (LC) separations is described. The device comprises an array of functional elements to carry out LC separations, integrates a novel microchip-MS interface to facilitate the orthogonal transposition of the microfluidic LC channel into an array of reservoirs, and enables sensitive MALDI-MS detection directly from the chip. Essentially, the device provides a snapshot MALDI-MS map of the content of the separation channel present on the chip. The detection of proteins with biomarker potential from MCF10A breast epithelial cell extracts, and detection limits in the low fmol range, are demonstrated. In addition, the design of the novel LC-MALDI-MS chip entices the promotion of a new concept for performing sample separations within the limited time-frame that accompanies the dead-volume of a separation channel. PMID:23592150

  13. Engineering on-chip nanoporous gold material libraries via precision photothermal treatment

    NASA Astrophysics Data System (ADS)

    Chapman, Christopher A. R.; Wang, Ling; Biener, Juergen; Seker, Erkin; Biener, Monika M.; Matthews, Manyalibo J.

    2015-12-01

    Libraries of nanostructured materials on a single chip are a promising platform for high throughput and combinatorial studies of structure-property relationships in the fields of physics and biology. Nanoporous gold (np-Au), produced by an alloy corrosion process, is a nanostructured material specifically suited for such studies because of its self-similar thermally induced coarsening behavior. However, traditional heat application techniques for the modification of np-Au are bulk processes that cannot be used to generate a library of different pore sizes on a single chip. Here, laser micro-processing offers an attractive solution to this problem by providing a means to apply energy with high spatial and temporal resolution. In the present study we use finite element multiphysics simulations to predict the effects of laser mode (continuous-wave vs. pulsed) and thermal conductivity of the supporting substrate on the local np-Au film temperatures during photothermal annealing. Based on these results we discuss the mechanisms by which the np-Au network is coarsened. Thermal transport simulations predict that continuous-wave mode laser irradiation of np-Au thin films on a silicon substrate supports the widest range of morphologies that can be created through photothermal annealing of np-Au. Using the guidance provided by simulations, we successfully fabricate an on-chip material library consisting of 81 np-Au samples of 9 different morphologies for use in the parallel study of structure-property relationships.Libraries of nanostructured materials on a single chip are a promising platform for high throughput and combinatorial studies of structure-property relationships in the fields of physics and biology. Nanoporous gold (np-Au), produced by an alloy corrosion process, is a nanostructured material specifically suited for such studies because of its self-similar thermally induced coarsening behavior. However, traditional heat application techniques for the modification of np-Au are bulk processes that cannot be used to generate a library of different pore sizes on a single chip. Here, laser micro-processing offers an attractive solution to this problem by providing a means to apply energy with high spatial and temporal resolution. In the present study we use finite element multiphysics simulations to predict the effects of laser mode (continuous-wave vs. pulsed) and thermal conductivity of the supporting substrate on the local np-Au film temperatures during photothermal annealing. Based on these results we discuss the mechanisms by which the np-Au network is coarsened. Thermal transport simulations predict that continuous-wave mode laser irradiation of np-Au thin films on a silicon substrate supports the widest range of morphologies that can be created through photothermal annealing of np-Au. Using the guidance provided by simulations, we successfully fabricate an on-chip material library consisting of 81 np-Au samples of 9 different morphologies for use in the parallel study of structure-property relationships. Electronic supplementary information (ESI) available: Details of sample preparation, fabrication of material libraries, as well as further analysis and supporting scanning electron micrographs can be found in ESI. See DOI: 10.1039/c5nr04580k

  14. Microstructural characterization of Ti-6Al-4V metal chips by focused ion beam (FIB) and transmission electron microscopy (TEM)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Schneider, Judy; Dong, Lei; Howe, Jane Y

    2011-01-01

    The microstructure of the secondary deformation zone (SDZ) near the cutting surface in metal chips of Ti-6Al-4V formed during machining was investigated using focused ion beam (FIB) specimen preparation and transmission electron microscopy (TEM) imaging. Use of the FIB allowed precise extraction of the specimen across this region to reveal its inhomogeneous microstructure resulting from the non-uniform distribution of strain, strain rate, and temperature generated during the cutting process. Initial imaging from conventional TEM foil preparation revealed microstructures ranging from heavily textured to regions of fine grains. Using FIB preparation, the transverse microstructure could be interpreted as fine grains nearmore » the cutting surface which transitioned to coarse grains toward the free surface. At the cutting surface a 10 nm thick recrystallized layer was observed capping a 20 nm thick amorphous layer.« less

  15. Effect of neodymium substitution on the electric and dielectric properties of Mn-Ni-Zn ferrite

    NASA Astrophysics Data System (ADS)

    Agami, W. R.

    2018-04-01

    Ferrite samples of Mn0.5Ni0.1Zn0.4NdxFe2-xO4 (x = 0.0, 0.01, 0.02, 0.05, 0.075 and 0.1) have been prepared by usual ceramic method. The temperature and composition dependences of the dc electric resistivity (ρdc) were studied. The frequency and composition dependences of the ac electric resistivity (ρac) and dielectric parameters (dielectric constant ε' and dielectric loss ε'') have been investigated. ρdc was found to decrease with temperature for all samples while it increases with increasing Nd3+ concentration. On the other hand, ρac and the dielectric properties were found to decrease with increasing the frequency while ρac increases and both ε' and ε'' decrease with increasing Nd3+ concentration. These results were explained by the Maxwell-Wagner two-layer model and Koops's theory. The improvement in dc and ac electric resistivities shows that these prepared materials are valid for decreasing the eddy current losses at high frequencies, so they can be used in the fabrication of multilayer chip inductor (MLCI) devices.

  16. Digital LAMP in a sample self-digitization (SD) chip

    PubMed Central

    Herrick, Alison M.; Dimov, Ivan K.; Lee, Luke P.; Chiu, Daniel T.

    2012-01-01

    This paper describes the realization of digital loop-mediated DNA amplification (dLAMP) in a sample self-digitization (SD) chip. Digital DNA amplification has become an attractive technique to quantify absolute concentrations of DNA in a sample. While digital polymerase chain reaction is still the most widespread implementation, its use in resource—limited settings is impeded by the need for thermal cycling and robust temperature control. In such situations, isothermal protocols that can amplify DNA or RNA without thermal cycling are of great interest. Here, we showed the successful amplification of single DNA molecules in a stationary droplet array using isothermal digital loop-mediated DNA amplification. Unlike most (if not all) existing methods for sample discretization, our design allows for automated, loss-less digitization of sample volumes on-chip. We demonstrated accurate quantification of relative and absolute DNA concentrations with sample volumes of less than 2 μl. We assessed the homogeneity of droplet size during sample self-digitization in our device, and verified that the size variation was small enough such that straightforward counting of LAMP-active droplets sufficed for data analysis. We anticipate that the simplicity and robustness of our SD chip make it attractive as an inexpensive and easy-to-operate device for DNA amplification, for example in point-of-care settings. PMID:22399016

  17. Consumption study and identification of methyl salicylate in spicy cassava chips

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nirjana, Marlene, E-mail: marlenenirjana@gmail.com; Anggadiredja, Kusnandar; Damayanti, Sophi

    Spicy cassava chips is a popular snack. However, some news in electronic media reported addition of balsam which is a banned food additives in that product to give extra spicy flavor. This study aimed to determine ITB students’ pattern of consumption, health problems caused by spicy chips consumption, and knowledge about illicit use of food additives in that product, and identify the main content of balsam namely methyl salicylate in 10 samples of spicy cassava chips taken from inside and outside about ITB campus. A total of 300 questionnaires distributed to ITB students then data processing was performed. Spicy cassavamore » chips sample macerated in 50 mL of methanol for 24 hours at room temperature, filtered and analyzed using gas chromatography capillary column with OV-1, nitrogen carrier gas and flame ionization detector. Based on questionnaires, 292 (97%) of 300 respondents had consumed spicy chips. A total of 247 (85%) from 292 respondents spicy chips consumed less than 3 times a week. A total of 195 respondents (67%) had experienced health problems after eating spicy chips. There were 137 (47%) of the 292 respondents who knew about the illicit addition of food additives into spicy chips; only 35 respondents (12%) who knew about balsam’s addition. There were 126 respondents (43%) who did not pay attention to their health because they will keep eating spicy chips despite the addition of banned food additives. Through the verification of the standard addition method in gas chromatography system with a hydrogen pressure of 1.5 bar, injector temperature 200 °C, detector temperature 230 °C, oven temperature 60 °C for 2 minutes and then increased to 230 °C with rate 6 °C/menit; linearity, limit of detection, limit of quantitation, accuracy, precision, and specificity parameters met the acceptance limits. From 10 spicy cassava chips samples which were analyzed, they did not reveal any content of methyl salicylate. Methyl salicylate contained in the positive control was 1.273 mg/mL.« less

  18. Potentiometric chip-based multipumping flow system for the simultaneous determination of fluoride, chloride, pH, and redox potential in water samples.

    PubMed

    Chango, Gabriela; Palacio, Edwin; Cerdà, Víctor

    2018-08-15

    A simple potentiometric chip-based multipumping flow system (MPFS) has been developed for the simultaneous determination of fluoride, chloride, pH, and redox potential in water samples. The proposed system was developed by using a poly(methyl methacrylate) chip microfluidic-conductor using the advantages of flow techniques with potentiometric detection. For this purpose, an automatic system has been designed and built by optimizing the variables involved in the process, such as: pH, ionic strength, stirring and sample volume. This system was applied successfully to water samples getting a versatile system with an analysis frequency of 12 samples per hour. Good correlation between chloride and fluoride concentration measured with ISE and ionic chromatography technique suggests satisfactory reliability of the system. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Lensfree fluorescent on-chip imaging of transgenic Caenorhabditis elegans over an ultra-wide field-of-view.

    PubMed

    Coskun, Ahmet F; Sencan, Ikbal; Su, Ting-Wei; Ozcan, Aydogan

    2011-01-06

    We demonstrate lensfree on-chip fluorescent imaging of transgenic Caenorhabditis elegans (C. elegans) over an ultra-wide field-of-view (FOV) of e.g., >2-8 cm(2) with a spatial resolution of ∼10 µm. This is the first time that a lensfree on-chip platform has successfully imaged fluorescent C. elegans samples. In our wide-field lensfree imaging platform, the transgenic samples are excited using a prism interface from the side, where the pump light is rejected through total internal reflection occurring at the bottom facet of the substrate. The emitted fluorescent signal from C. elegans samples is then recorded on a large area opto-electronic sensor-array over an FOV of e.g., >2-8 cm(2), without the use of any lenses, thin-film interference filters or mechanical scanners. Because fluorescent emission rapidly diverges, such lensfree fluorescent images recorded on a chip look blurred due to broad point-spread-function of our platform. To combat this resolution challenge, we use a compressive sampling algorithm to uniquely decode the recorded lensfree fluorescent patterns into higher resolution images, demonstrating ∼10 µm resolution. We tested the efficacy of this compressive decoding approach with different types of opto-electronic sensors to achieve a similar resolution level, independent of the imaging chip. We further demonstrate that this wide FOV lensfree fluorescent imaging platform can also perform sequential bright-field imaging of the same samples using partially-coherent lensfree digital in-line holography that is coupled from the top facet of the same prism used in fluorescent excitation. This unique combination permits ultra-wide field dual-mode imaging of C. elegans on a chip which could especially provide a useful tool for high-throughput screening applications in biomedical research.

  20. A scalable self-priming fractal branching microchannel net chip for digital PCR.

    PubMed

    Zhu, Qiangyuan; Xu, Yanan; Qiu, Lin; Ma, Congcong; Yu, Bingwen; Song, Qi; Jin, Wei; Jin, Qinhan; Liu, Jinyu; Mu, Ying

    2017-05-02

    As an absolute quantification method at the single-molecule level, digital PCR has been widely used in many bioresearch fields, such as next generation sequencing, single cell analysis, gene editing detection and so on. However, existing digital PCR methods still have some disadvantages, including high cost, sample loss, and complicated operation. In this work, we develop an exquisite scalable self-priming fractal branching microchannel net digital PCR chip. This chip with a special design inspired by natural fractal-tree systems has an even distribution and 100% compartmentalization of the sample without any sample loss, which is not available in existing chip-based digital PCR methods. A special 10 nm nano-waterproof layer was created to prevent the solution from evaporating. A vacuum pre-packaging method called self-priming reagent introduction is used to passively drive the reagent flow into the microchannel nets, so that this chip can realize sequential reagent loading and isolation within a couple of minutes, which is very suitable for point-of-care detection. When the number of positive microwells stays in the range of 100 to 4000, the relative uncertainty is below 5%, which means that one panel can detect an average of 101 to 15 374 molecules by the Poisson distribution. This chip is proved to have an excellent ability for single molecule detection and quantification of low expression of hHF-MSC stem cell markers. Due to its potential for high throughput, high density, low cost, lack of sample and reagent loss, self-priming even compartmentalization and simple operation, we envision that this device will significantly expand and extend the application range of digital PCR involving rare samples, liquid biopsy detection and point-of-care detection with higher sensitivity and accuracy.

  1. Lensfree Fluorescent On-Chip Imaging of Transgenic Caenorhabditis elegans Over an Ultra-Wide Field-of-View

    PubMed Central

    Ozcan, Aydogan

    2011-01-01

    We demonstrate lensfree on-chip fluorescent imaging of transgenic Caenorhabditis elegans (C. elegans) over an ultra-wide field-of-view (FOV) of e.g., >2–8 cm2 with a spatial resolution of ∼10µm. This is the first time that a lensfree on-chip platform has successfully imaged fluorescent C. elegans samples. In our wide-field lensfree imaging platform, the transgenic samples are excited using a prism interface from the side, where the pump light is rejected through total internal reflection occurring at the bottom facet of the substrate. The emitted fluorescent signal from C. elegans samples is then recorded on a large area opto-electronic sensor-array over an FOV of e.g., >2–8 cm2, without the use of any lenses, thin-film interference filters or mechanical scanners. Because fluorescent emission rapidly diverges, such lensfree fluorescent images recorded on a chip look blurred due to broad point-spread-function of our platform. To combat this resolution challenge, we use a compressive sampling algorithm to uniquely decode the recorded lensfree fluorescent patterns into higher resolution images, demonstrating ∼10 µm resolution. We tested the efficacy of this compressive decoding approach with different types of opto-electronic sensors to achieve a similar resolution level, independent of the imaging chip. We further demonstrate that this wide FOV lensfree fluorescent imaging platform can also perform sequential bright-field imaging of the same samples using partially-coherent lensfree digital in-line holography that is coupled from the top facet of the same prism used in fluorescent excitation. This unique combination permits ultra-wide field dual-mode imaging of C. elegans on a chip which could especially provide a useful tool for high-throughput screening applications in biomedical research. PMID:21253611

  2. Novel Non-Intrusive Trans-Dermal Remote Wireless Micro-Fluidic Monitoring System Applied to Continuous Glucose and Lactate Assays for Casualty Care and Combat Readiness Assessment

    DTIC Science & Technology

    2004-09-01

    identification of the lettered features. 2.2 BFIT Sampling Chip The BFIT sampling chip is a flexible patch-like chip with a multilayer polymeric metal...PPy) and glucose oxidase (GOD). The BFIT fabrication process uses SU8 as a principal structural material consisting of five steps (Figure 2). This...process is a subset of an earlier technology developed for the polymer material PDMS.11,12,13,14,15 The first step was the deposition of a Teflon

  3. Multichannel microfluidic chip for rapid and reliable trapping and imaging plant-parasitic nematodes

    NASA Astrophysics Data System (ADS)

    Amrit, Ratthasart; Sripumkhai, Witsaroot; Porntheeraphat, Supanit; Jeamsaksiri, Wutthinan; Tangchitsomkid, Nuchanart; Sutapun, Boonsong

    2013-05-01

    Faster and reliable testing technique to count and identify nematode species resided in plant roots is therefore essential for export control and certification. This work proposes utilizing a multichannel microfluidic chip with an integrated flow-through microfilter to retain the nematodes in a trapping chamber. When trapped, it is rather simple and convenient to capture images of the nematodes and later identify their species by a trained technician. Multiple samples can be tested in parallel using the proposed microfluidic chip therefore increasing number of samples tested per day.

  4. Dentine chips produced by nickel-titanium rotary instruments.

    PubMed

    Guppy, D R; Curtis, R V; Ford, T R

    2000-12-01

    This study aimed to compare the cross-sectional shape of two nickel-titanium rotary instruments, namely ProFile and Quantec files, both ISO 25, 0.06 taper, and sought to relate this to the chips produced by cutting dentine. A limited comparison was made with stainless steel engine reamers. First, five files of each type were sectioned transversely at 12 mm, 8 mm and 4 mm from the tip and examined by scanning electron microscopy. The cutting angles were assessed by a direct measurement technique which allowed for the inclination of a cutting edge to the root canal. Second, eight samples of cutting debris were collected from instrumentation by each type of nickel-titanium file and four samples from the engine reamers. The major and minor axis, area and roundness of the dentine chips in each sample were measured using computerized particle analysis. The results demonstrated that all files had a negative cutting angle which varied at the different levels (ProFiles range 69.4 degrees to 58.4 degrees and Quantec range 74.8 degrees to 56.8 degrees). The consistency within files of the same type was good as demonstrated by low standard deviations, except for Quantec files at the 4 mm level where higher standard deviations of 4.1 degrees and 5.5 degrees for the two blades were found. The chip analysis showed significant differences between chips produced by ProFile and Quantec files (P < 0.05). The latter were larger and rounder. The chips from the ProFile and the engine reamer chips were similar in dimension (P > 0.05). No simple relationship existed between file geometry and the dentine chips produced during instrumentation.

  5. Hydrothermal carbonization for the preparation of hydrochars from glucose, cellulose, chitin, chitosan and wood chips via low-temperature and their characterization.

    PubMed

    Simsir, Hamza; Eltugral, Nurettin; Karagoz, Selhan

    2017-12-01

    In this work, the hydrothermal carbonization of glucose, cellulose, chitin, chitosan and wood chips at 200°C at processing times between 6 and 48h was studied. The carbonization degree of wood chips, cellulose and chitosan obviously increases as function of time. The heating value of glucose increases to 88% upon carbonization for 48h, while it is only 5% for chitin. It is calculated to be between 44 and 73% for wood chips, chitosan and cellulose. Glucose yielded complete formation of spherical hydrochar structures at a shorter processing time, as low as 12h. However, carbon spheres with narrow size (∼560nm) distribution were obtained upon 48h of residence time. Cellulose and wood chips yielded a similar morphology with an irregular size distribution. Chitin seemed not to undergo hydrothermal carbonization, whereas densely aggregated spheres of a uniform size around 42nm were obtained from chitosan after 18h. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. On-chip concentration of bacteria using a 3D dielectrophoretic chip and subsequent laser-based DNA extraction in the same chip

    NASA Astrophysics Data System (ADS)

    Cho, Yoon-Kyoung; Kim, Tae-hyeong; Lee, Jeong-Gun

    2010-06-01

    We report the on-chip concentration of bacteria using a dielectrophoretic (DEP) chip with 3D electrodes and subsequent laser-based DNA extraction in the same chip. The DEP chip has a set of interdigitated Au post electrodes with 50 µm height to generate a network of non-uniform electric fields for the efficient trapping by DEP. The metal post array was fabricated by photolithography and subsequent Ni and Au electroplating. Three model bacteria samples (Escherichia coli, Staphylococcus epidermidis, Streptococcus mutans) were tested and over 80-fold concentrations were achieved within 2 min. Subsequently, on-chip DNA extraction from the concentrated bacteria in the 3D DEP chip was performed by laser irradiation using the laser-irradiated magnetic bead system (LIMBS) in the same chip. The extracted DNA was analyzed with silicon chip-based real-time polymerase chain reaction (PCR). The total process of on-chip bacteria concentration and the subsequent DNA extraction can be completed within 10 min including the manual operation time.

  7. Sputtered Pd as hydrogen storage for a chip-integrated microenergy system.

    PubMed

    Slavcheva, E; Ganske, G; Schnakenberg, U

    2014-01-01

    The work presents a research on preparation and physical and electrochemical characterisation of dc magnetron sputtered Pd films envisaged for application as hydrogen storage in a chip-integrated hydrogen microenergy system. The influence of the changes in the sputtering pressure on the surface structure, morphology, and roughness was analysed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AMF). The electrochemical activity towards hydrogen adsorption/desorption and formation of PdH were investigated in 0.5 M H2SO4 using the methods of cyclic voltammetry and galvanostatic polarisation. The changes in the electrical properties of the films as a function of the sputtering pressure and the level of hydrogenation were evaluated before and immediately after the electrochemical charging tests, using a four-probe technique. The research resulted in establishment of optimal sputter regime, ensuring fully reproducible Pd layers with highly developed surface, moderate porosity, and mechanical stability. Selected samples were integrated as hydrogen storage in a newly developed unitized microenergy system and tested in charging (water electrolysis) and discharging (fuel cell) operative mode at ambient conditions demonstrating a stable recycling performance.

  8. Optical ammonia gas sensor based on a porous silicon rugate filter coated with polymer-supported dye.

    PubMed

    Shang, Yunling; Wang, Xiaobo; Xu, Erchao; Tong, Changlun; Wu, Jianmin

    2011-01-24

    An ammonia gas sensor chip was prepared by coating an electrochemically-etched porous Si rugate filter with a chitosan film that is crosslinked by glycidoxypropyltrimethoxysilane (GPTMS). The bromothylmol blue (BTB), a pH indicator, was loaded in the film as ammonia-sensing molecules. White light reflected from the porous Si has a narrow bandwidth spectrum with a peak at 610 nm. Monitoring reflective optical intensity at the peak position allows for direct, real-time observation of changes in the concentration of ammonia gas in air samples. The reflective optical intensity decreased linearly with increasing concentrations of ammonia gas over the range of 0-100 ppm. The lowest detection limit was 0.5 ppm for ammonia gas. At optimum conditions, the full response time of the ammonia gas sensor was less than 15s. The sensor chip also exhibited a good long-term stability over 1 year. Therefore, the simple sensor design has potential application in miniaturized optical measurement for online ammonia gas detection. Copyright © 2010 Elsevier B.V. All rights reserved.

  9. A lab-on-chip for biothreat detection using single-molecule DNA mapping.

    PubMed

    Meltzer, Robert H; Krogmeier, Jeffrey R; Kwok, Lisa W; Allen, Richard; Crane, Bryan; Griffis, Joshua W; Knaian, Linda; Kojanian, Nanor; Malkin, Gene; Nahas, Michelle K; Papkov, Vyacheslav; Shaikh, Saad; Vyavahare, Kedar; Zhong, Qun; Zhou, Yi; Larson, Jonathan W; Gilmanshin, Rudolf

    2011-03-07

    Rapid, specific, and sensitive detection of airborne bacteria, viruses, and toxins is critical for biodefense, yet the diverse nature of the threats poses a challenge for integrated surveillance, as each class of pathogens typically requires different detection strategies. Here, we present a laboratory-on-a-chip microfluidic device (LOC-DLA) that integrates two unique assays for the detection of airborne pathogens: direct linear analysis (DLA) with unsurpassed specificity for bacterial threats and Digital DNA for toxins and viruses. The LOC-DLA device also prepares samples for analysis, incorporating upstream functions for concentrating and fractionating DNA. Both DLA and Digital DNA assays are single molecule detection technologies, therefore the assay sensitivities depend on the throughput of individual molecules. The microfluidic device and its accompanying operation protocols have been heavily optimized to maximize throughput and minimize the loss of analyzable DNA. We present here the design and operation of the LOC-DLA device, demonstrate multiplex detection of rare bacterial targets in the presence of 100-fold excess complex bacterial mixture, and demonstrate detection of picogram quantities of botulinum toxoid.

  10. Microfluidic approaches to malaria detection

    PubMed Central

    Gascoyne, Peter; Satayavivad, Jutamaad; Ruchirawat, Mathuros

    2009-01-01

    Microfluidic systems are under development to address a variety of medical problems. Key advantages of micrototal analysis systems based on microfluidic technology are the promise of small size and the integration of sample handling and measurement functions within a single, automated device having low mass-production costs. Here, we review the spectrum of methods currently used to detect malaria, consider their advantages and disadvantages, and discuss their adaptability towards integration into small, automated micro total analysis systems. Molecular amplification methods emerge as leading candidates for chip-based systems because they offer extremely high sensitivity, the ability to recognize malaria species and strain, and they will be adaptable to the detection of new genotypic signatures that will emerge from current genomic-based research of the disease. Current approaches to the development of chip-based molecular amplification are considered with special emphasis on flow-through PCR, and we present for the first time the method of malaria specimen preparation by dielectrophoretic field-flow-fractionation. Although many challenges must be addressed to realize a micrototal analysis system for malaria diagnosis, it is concluded that the potential benefits of the approach are well worth pursuing. PMID:14744562

  11. Electrochemical pesticide detection with AutoDip--a portable platform for automation of crude sample analyses.

    PubMed

    Drechsel, Lisa; Schulz, Martin; von Stetten, Felix; Moldovan, Carmen; Zengerle, Roland; Paust, Nils

    2015-02-07

    Lab-on-a-chip devices hold promise for automation of complex workflows from sample to answer with minimal consumption of reagents in portable devices. However, complex, inhomogeneous samples as they occur in environmental or food analysis may block microchannels and thus often cause malfunction of the system. Here we present the novel AutoDip platform which is based on the movement of a solid phase through the reagents and sample instead of transporting a sequence of reagents through a fixed solid phase. A ball-pen mechanism operated by an external actuator automates unit operations such as incubation and washing by consecutively dipping the solid phase into the corresponding liquids. The platform is applied to electrochemical detection of organophosphorus pesticides in real food samples using an acetylcholinesterase (AChE) biosensor. Minimal sample preparation and an integrated reagent pre-storage module hold promise for easy handling of the assay. Detection of the pesticide chlorpyrifos-oxon (CPO) spiked into apple samples at concentrations of 10(-7) M has been demonstrated. This concentration is below the maximum residue level for chlorpyrifos in apples defined by the European Commission.

  12. Efforts to Improve School Lunch Programs--Are They Paying Off?

    DTIC Science & Technology

    1981-09-09

    ingredients than other districts that purchase more ready-made lunch components. --The amount of special equipment (warming lights, dough mixers, bun slicers...components such as burritos, pizza, or peanut butter cookies . Very few lunch components were significantly short weight and, in all cases, shortages were...fruit and/or juice; potato chips, corn chips, or cookie ; and milk. All food served in the lunch program is prepared in the cafeterias. Nutrient-based

  13. Hand drawing of pencil electrodes on paper platforms for contactless conductivity detection of inorganic cations in human tear samples using electrophoresis chips.

    PubMed

    Chagas, Cyro L S; Costa Duarte, Lucas; Lobo-Júnior, Eulício O; Piccin, Evandro; Dossi, Nicolò; Coltro, Wendell K T

    2015-08-01

    This paper describes for the first time the fabrication of pencil drawn electrodes (PDE) on paper platforms for capacitively coupled contactless conductivity detection (C(4) D) on electrophoresis microchips. PDE-C(4) D devices were attached on PMMA electrophoresis chips and used for detection of K(+) and Na(+) in human tear samples. PDE-C(4) D devices were produced on office paper and chromatographic paper platforms and their performance were thoroughly investigated using a model mixture containing K(+) , Na(+) , and Li(+) . In comparison with chromatographic paper, PDE-C(4) D fabricated on office paper has exhibited better performance due to its higher electrical conductivity. Furthermore, the detector response was similar to that recorded with electrodes prepared with copper adhesive tape. The fabrication of PDE-C(4) D on office paper has offered great advantages including extremely low cost (< $ 0.004 per unit), reduced fabrication time (< 5 min), and minimal instrumentation (pencil and paper). The proposed electrodes demonstrated excellent analytical performance with good reproducibility. For an inter-PDE comparison (n = 7), the RSD values for migration time, peak area, and separation efficiency were lower than 2.5, 10.5, and 14%, respectively. The LOD's achieved for K(+) , Na(+) , and Li(+) were 4.9, 6.8, and 9.0 μM, respectively. The clinical feasibility of the proposed approach was successfully demonstrated with the quantitative analysis of K(+) and Na(+) in tear samples. The concentration levels found for K(+) and Na(+) were, respectively, 20.8 ± 0.1 mM and 101.2 ± 0.1 mM for sample #1, and 20.4 ± 0.1 mM and 111.4 ± 0.1 mM for sample #2. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Development of size reduction equations for calculating power input for grinding pine wood chips using hammer mill

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Naimi, Ladan J.; Collard, Flavien; Bi, Xiaotao

    Size reduction is an unavoidable operation for preparing biomass for biofuels and bioproduct conversion. Yet, there is considerable uncertainty in power input requirement and the uniformity of ground biomass. Considerable gains are possible if the required power input for a size reduction ratio is estimated accurately. In this research three well-known mechanistic equations attributed to Rittinger, Kick, and Bond available for predicting energy input for grinding pine wood chips were tested against experimental grinding data. Prior to testing, samples of pine wood chips were conditioned to 11.7% wb, moisture content. The wood chips were successively ground in a hammer millmore » using screen sizes of 25.4 mm, 10 mm, 6.4 mm, and 3.2 mm. The input power and the flow of material into the grinder were recorded continuously. The recorded power input vs. mean particle size showed that the Rittinger equation had the best fit to the experimental data. The ground particle sizes were 4 to 7 times smaller than the size of installed screen. Geometric mean size of particles were calculated using two methods (1) Tyler sieves and using particle size analysis and (2) Sauter mean diameter calculated from the ratio of volume to surface that were estimated from measured length and width. The two mean diameters agreed well, pointing to the fact that either mechanical sieving or particle imaging can be used to characterize particle size. In conclusion, specific energy input to the hammer mill increased from 1.4 kWh t –1 (5.2 J g –1) for large 25.1-mm screen to 25 kWh t –1 (90.4 J g –1) for small 3.2-mm screen.« less

  15. Shape-anchored porous polymer monoliths for integrated online solid-phase extraction-microchip electrophoresis-electrospray ionization mass spectrometry.

    PubMed

    Nordman, Nina; Barrios-Lopez, Brianda; Laurén, Susanna; Suvanto, Pia; Kotiaho, Tapio; Franssila, Sami; Kostiainen, Risto; Sikanen, Tiina

    2015-02-01

    We report a simple protocol for fabrication of shape-anchored porous polymer monoliths (PPMs) for on-chip SPE prior to online microchip electrophoresis (ME) separation and on-chip (ESI/MS). The chip design comprises a standard ME separation channel with simple cross injector and a fully integrated ESI emitter featuring coaxial sheath liquid channel. The monolith zone was prepared in situ at the injection cross by laser-initiated photopolymerization through the microchip cover layer. The use of high-power laser allowed not only maskless patterning of a precisely defined monolith zone, but also faster exposure time (here, 7 min) compared with flood exposure UV lamps. The size of the monolith pattern was defined by the diameter of the laser output (∅500 μm) and the porosity was geared toward high through-flow to allow electrokinetic actuation and thus avoid coupling to external pumps. Placing the monolith at the injection cross enabled firm anchoring based on its cross-shape so that no surface premodification with anchoring linkers was needed. In addition, sample loading and subsequent injection (elution) to the separation channel could be performed similar to standard ME setup. As a result, 15- to 23-fold enrichment factors were obtained already at loading (preconcentration) times as short as 25 s without sacrificing the throughput of ME analysis. The performance of the SPE-ME-ESI/MS chip was repeatable within 3.1% and 11.5% RSD (n = 3) in terms of migration time and peak height, respectively, and linear correlation was observed between the loading time and peak area. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Development of size reduction equations for calculating power input for grinding pine wood chips using hammer mill

    DOE PAGES

    Naimi, Ladan J.; Collard, Flavien; Bi, Xiaotao; ...

    2016-01-05

    Size reduction is an unavoidable operation for preparing biomass for biofuels and bioproduct conversion. Yet, there is considerable uncertainty in power input requirement and the uniformity of ground biomass. Considerable gains are possible if the required power input for a size reduction ratio is estimated accurately. In this research three well-known mechanistic equations attributed to Rittinger, Kick, and Bond available for predicting energy input for grinding pine wood chips were tested against experimental grinding data. Prior to testing, samples of pine wood chips were conditioned to 11.7% wb, moisture content. The wood chips were successively ground in a hammer millmore » using screen sizes of 25.4 mm, 10 mm, 6.4 mm, and 3.2 mm. The input power and the flow of material into the grinder were recorded continuously. The recorded power input vs. mean particle size showed that the Rittinger equation had the best fit to the experimental data. The ground particle sizes were 4 to 7 times smaller than the size of installed screen. Geometric mean size of particles were calculated using two methods (1) Tyler sieves and using particle size analysis and (2) Sauter mean diameter calculated from the ratio of volume to surface that were estimated from measured length and width. The two mean diameters agreed well, pointing to the fact that either mechanical sieving or particle imaging can be used to characterize particle size. In conclusion, specific energy input to the hammer mill increased from 1.4 kWh t –1 (5.2 J g –1) for large 25.1-mm screen to 25 kWh t –1 (90.4 J g –1) for small 3.2-mm screen.« less

  17. Effects of calcium supplements on the quality and acrylamide content of puffed shrimp chips.

    PubMed

    Chen, Tai-Yuan; Luo, Hsuan-Min; Hsu, Pang-Hung; Sung, Wen-Chieh

    2016-01-01

    The quality and acrylamide content of deep-fried and microwave-puffed shrimp chips fortified with 0.1%, 0.5%, or 1.0% calcium salts (calcium lactate, calcium carbonate, calcium citrate, or calcium acetate) were investigated. Microwave-puffed shrimp chips contained higher amounts of acrylamide (130.43 ppb) than did deep-fried shrimp chips. The greatest mitigation of acrylamide formation in overfried chips was obtained with 0.1% calcium lactate. All browning indexes of fortified shrimp chips, whether deep-fried or microwave-puffed, were reduced. L* values of microwave-puffed shrimp chips were higher than those of deep-fried shrimp chips, whereas a* and b* values and browning indexes were lower. Color differences (ΔE) between deep-fried puffed shrimp chips fortified with calcium salts and a control sample were higher than 5, and the sensory scores of shrimp chips were significantly decreased by the addition of calcium lactate. Copyright © 2015. Published by Elsevier B.V.

  18. Comparison of edge chipping resistance of PFM and veneered zirconia specimens

    PubMed Central

    Quinn, Janet B.; Sundar, Veeraraghavan; Parry, Edward E.; Quinn, George D.

    2011-01-01

    Objectives To investigate the chipping resistance of veneered zirconia specimens and compare it to the chipping resistance of porcelain fused to metal (PFM) specimens. Methods Veneered zirconia and PFM bar specimens were prepared in clinically relevant thicknesses. The specimen edges were chipped with different magnitude forces, producing chips of various sizes. The range of sizes included small chips that did not penetrate all the way through the veneers to the substrates, and also chips that were very large and reached the zirconia or metal substrates. The relationship between force magnitude and chip size (edge distance) was graphed. The resulting curves were compared for the veneered zirconia and PFM specimens. Knoop hardness vs. force graphs for the veneers and substrates were also obtained. Results The zirconia and PFM veneer chipping data followed a power law (coefficient of determination, R2 > 0.93) as expected from the literature. The curves overlapped within the combined data scatter, indicating similar resistance to chipping. The chips made in both types of specimens detached and did not penetrate into the substrate when they reached the veneer/substrate intersections. The hardness–load curves for the veneers and substrates all exhibited an indentation size effect (ISE) at low loads. The Knoop hardness values with uncertainties of ±one standard deviation at 4 N loads for the metal, zirconia, and the metal and zirconia veneers are: (2.02 ± 0.08, 12.01 ± 0.39, 4.24 ± 0.16 and 4.36 ± 0.02 GPa), respectively, with no statistically significant difference between the veneers (Tukey pairwise comparison at 0.95 family confidence). Significance This work indicates that a similar resistance to chipping might be expected for veneered zirconia and PFM restorations, in spite of the large difference in substrate hardness. Differences in susceptibility to chip spalling were not detected, but the chips in both specimen types detached off the sides in a similar manner instead of extending into the substrates. PMID:19748115

  19. A piezo-ring-on-chip microfluidic device for simple and low-cost mass spectrometry interfacing.

    PubMed

    Tsao, Chia-Wen; Lei, I-Chao; Chen, Pi-Yu; Yang, Yu-Liang

    2018-02-12

    Mass spectrometry (MS) interfacing technology provides the means for incorporating microfluidic processing with post MS analysis. In this study, we propose a simple piezo-ring-on-chip microfluidic device for the controlled spraying of MALDI-MS targets. This device uses a low-cost, commercially-available ring-shaped piezoelectric acoustic atomizer (piezo-ring) directly integrated into a polydimethylsiloxane microfluidic device to spray the sample onto the MS target substrate. The piezo-ring-on-chip microfluidic device's design, fabrication, and actuation, and its pulsatile pumping effects were evaluated. The spraying performance was examined by depositing organic matrix samples onto the MS target substrate by using both an automatic linear motion motor, and manual deposition. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was performed to analyze the peptide samples on the MALDI target substrates. Using our technique, model peptides with 10 -6 M concentration can be successfully detected. The results also indicate that the piezo-ring-on-chip approach forms finer matrix crystals and presents better MS signal uniformity with little sample consumption compared to the conventional pipetting method.

  20. A single-chip 32-channel analog beamformer with 4-ns delay resolution and 768-ns maximum delay range for ultrasound medical imaging with a linear array transducer.

    PubMed

    Um, Ji-Yong; Kim, Yoon-Jee; Cho, Seong-Eun; Chae, Min-Kyun; Kim, Byungsub; Sim, Jae-Yoon; Park, Hong-June

    2015-02-01

    A single-chip 32-channel analog beamformer is proposed. It achieves a delay resolution of 4 ns and a maximum delay range of 768 ns. It has a focal-point based architecture, which consists of 7 sub-analog beamformers (sub-ABF). Each sub-ABF performs a RX focusing operation for a single focal point. Seven sub-ABFs perform a time-interleaving operation to achieve the maximum delay range of 768 ns. Phase interpolators are used in sub-ABFs to generate sampling clocks with the delay resolution of 4 ns from a low frequency system clock of 5 MHz. Each sub-ABF samples 32 echo signals at different times into sampling capacitors, which work as analog memory cells. The sampled 32 echo signals of each sub-ABF are originated from one target focal point at one instance. They are summed at one instance in a sub-ABF to perform the RX focusing for the target focal point. The proposed ABF chip has been fabricated in a 0.13- μ m CMOS process with an active area of 16 mm (2). The total power consumption is 287 mW. In measurement, the digital echo signals from a commercial ultrasound medical imaging machine were applied to the fabricated chip through commercial DAC chips. Due to the speed limitation of the DAC chips, the delay resolution was relaxed to 10 ns for the real-time measurement. A linear array transducer with no steering operation is used in this work.

  1. Diagnostic value of protein chips constructed by lung-cancer-associated markers selected by the T7 phage display library.

    PubMed

    Li, Hong-Mei; Guo, Kang; Yu, Zhuang; Feng, Rui; Xu, Ping

    2015-07-01

    Traditional diagnostic technology with tumor biomarkers is inefficient, expensive and requires a large number of serum samples. The purpose of this study was to construct human lung cancer protein chips with new lung cancer biomarkers screened by the T7-phage display library, and improve the early diagnosis rate of lung cancer. A T7-phage cDNA display library was constructed of fresh samples from 30 lung cancer patients. With biopanning and high-throughput screening, we gained the immunogenic phage clones from the cDNA library. The insert of selected phage was blasted at GeneBank for alignment to find the exact or the most similar known genes. Protein chips were then constructed and used to assay their expression level in lung cancer serum from 217 cases of lung cancer groups:80 cases of benign lung disease and 220 healthy controls. After four rounds of Biopanning and two rounds of enzyme-linked immunosorbent assay, 12 phage monoclonal samples were selected from 2880 phage monoclonal samples. After blasting at GeneBank, six similar genes were used to construct diagnostic protein chips. The protein chips were then used to assay expression level in lung cancer serum. The expression level of six genes in lung cancer groups was significantly higher than those in the other two groups (P < 0.05). In this study, we successfully constructed diagnostic protein chips with biomarkers selected from the lung cancer T7-phage cDNA library, which can be used for the early screening of lung cancer patients.

  2. Low-Cost Chemical-Responsive Adhesive Sensing Chips.

    PubMed

    Tan, Weirui; Zhang, Liyuan; Shen, Wei

    2017-12-06

    Chemical-responsive adhesive sensing chip is a new low-cost analytical platform that uses adhesive tape loaded with indicator reagents to detect or quantify the target analytes by directly sticking the tape to the samples of interest. The chemical-responsive adhesive sensing chips can be used with paper to analyze aqueous samples; they can also be used to detect and quantify solid, particulate, and powder analytes. The colorimetric indicators become immediately visible as the contact between the functionalized adhesives and target samples is made. The chemical-responsive adhesive sensing chip expands the capability of paper-based analytical devices to analyze solid, particulate, or powder materials via one-step operation. It is also a simpler alternative way, to the covalent chemical modification of paper, to eliminate indicator leaching from the dipstick-style paper sensors. Chemical-responsive adhesive chips can display analytical results in the form of colorimetric dot patterns, symbols, and texts, enabling clear understanding of assay results by even nonprofessional users. In this work, we demonstrate the analyses of heavy metal salts in silica powder matrix, heavy metal ions in water, and bovine serum albumin in an aqueous solution. The detection is one-step, specific, sensitive, and easy-to-operate.

  3. Focusing analytes from 50 μL into 500 pL: On-chip focusing from large sample volumes using isotachophoresis.

    PubMed

    van Kooten, Xander F; Truman-Rosentsvit, Marianna; Kaigala, Govind V; Bercovici, Moran

    2017-09-05

    The use of on-chip isotachophoresis assays for diagnostic applications is often limited by the small volumes of standard microfluidic channels. Overcoming this limitation is particularly important for detection of 'discrete' biological targets (such as bacteria) at low concentrations, where the volume of processed liquid in a standard microchannel might not contain any targets. We present a novel microfluidic chip that enables ITP focusing of target analytes from initial sample volumes of 50 μL into a concentrated zone with a volume of 500 pL, corresponding to a 100,000-fold increase in mean concentration, and a 300,000-fold increase in peak concentration. We present design considerations for limiting sample dispersion in such large-volume focusing (LVF) chips and discuss the trade-off between assay time and Joule heating, which ultimately governs the scalability of LVF designs. Finally, we demonstrate a 100-fold improvement of ITP focusing performance in the LVF chip as compared to conventional microchannels, and apply this enhancement to achieve highly sensitive detection of both molecular targets (DNA, down to 10 fM) and whole bacteria (down to 100 cfu/mL).

  4. Enrichment and Detection of Escherichia coli O157:H7 from Water Samples Using an Antibody Modified Microfluidic Chip

    PubMed Central

    Dharmasiri, Udara; Witek, Małgorzata A.; Adams, Andre A.; Osiri, John K.; Hupert, Mateusz L.; Bianchi, Thomas S.; Roelke, Daniel L.; Soper, Steven A.

    2010-01-01

    Low abundant (<100 cells mL-1) E. coli O157:H7 cells were isolated and enriched from environmental water samples using a microfluidic chip. The poly(methylmethacrylate), PMMA, chip contained 8 devices each equipped with 16 curvilinear high aspect ratio channels that were covalently decorated with polyclonal anti-O157 antibodies (pAb) and could search for rare cells through a pAb mediated process. The chip could process independently 8 different samples or one sample using 8 different parallel inputs to increase volume processing throughput. After cell enrichment, cells were released and enumerated using bench top real-time quantitative PCR, targeting genes which effectively discriminated the O157:H7 serotype from other non-pathogenic bacteria. The recovery of target cells from water samples was determined to be ~72%, and the limit-of-detection was found to be 6 colony forming units (cfu) using the slt1 gene as a reporter. We subsequently performed analysis of lake and waste water samples. The simplicity in manufacturing and ease of operation makes this device attractive for the selection of pathogenic species from a variety of water supplies suspected of containing bacterial pathogens at extremely low frequencies. PMID:20218574

  5. 7 CFR 953.322 - Handling regulation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Agreements and Orders; Fruits, Vegetables, Nuts), DEPARTMENT OF AGRICULTURE IRISH POTATOES GROWN IN..., potatoes for dehydration, chips, shoestrings, starch, and flour. It includes only that preparation of...

  6. Preparation of a YAG:Ce phosphor glass by screen-printing technology and its application in LED packaging.

    PubMed

    Yang, Liang; Chen, Mingxiang; Lv, Zhicheng; Wang, Simin; Liu, Xiaogang; Liu, Sheng

    2013-07-01

    A simple and practical method for preparing phosphor glass is proposed. Phosphor distribution and element analysis are investigated by optical microscope and field emission scanning electron microscope (FE-SEM). The phosphor particles dispersed in the matrix are vividly observed, and their distributions are uniform. Spectrum distribution and color coordinates dependent on the thickness of the screen-printed phosphor layer coupled with a blue light emitting diode (LED) chip are studied. The luminous efficacy of the 75 μm printed phosphor-layer phosphor glass packaged white LED is 81.24 lm/W at 350 mA. This study opens up many possibilities for applications using the phosphor glass on a selected chip in which emission is well absorbed by all phosphors. The screen-printing technique also offers possibilities for the design and engineering of complex phosphor layers on glass substrates. Phosphor screen-printing technology allows the realization of high stability and thermal conductivity for the phosphor layer. This phosphor glass method provides many possibilities for LED packing, including thin-film flip chip and remote phosphor technology.

  7. InGaN-Based Light-Emitting Diodes Grown on a Micro/Nanoscale Hybrid Patterned Sapphire Substrate.

    PubMed

    Ke, Wen-Cheng; Lee, Fang-Wei; Chiang, Chih-Yung; Liang, Zhong-Yi; Chen, Wei-Kuo; Seong, Tae-Yeon

    2016-12-21

    A hybrid patterned sapphire substrate (hybrid-PSS) was prepared using an anodic aluminum oxide etching mask to transfer nanopatterns onto a conventional patterned sapphire substrate with microscale patterns (bare-PSS). The threading dislocation (TD) suppression of light-emitting diodes (LEDs) grown on a hybrid-PSS (HP-LED) exhibits a smaller reverse leakage current compared with that of LEDs grown on a bare-PSS (BP-LED). The strain-free GaN buffer layer and fully strained InGaN active layer were evidenced by cross-sectional Raman spectra and reciprocal space mapping of the X-ray diffraction intensity for both samples. The calculated piezoelectric fields for both samples are close, implying that the quantum-confined Stark effect was not a dominant mechanism influencing the electroluminescence (EL) peak wavelength under a high injection current. The bandgap shrinkage effect of the InGaN well layer was considered to explain the large red-shifted EL peak wavelength under high injection currents. The estimated LED chip temperatures rise from room temperature to 150 °C and 75 °C for BP-LED and HP-LED, respectively, at a 600-mA injection current. This smaller temperature rise of the LED chip is attributed to the increased contact area between the sapphire and the LED structural layer because of the embedded nanopattern. Although the chip generates more heat at high injection currents, the accumulated heat can be removed to outside the chip effectively. The high diffuse reflection (DR) rate of hybrid-PSS increases the escape probability of photons, resulting in an increase in the viewing angle of the LEDs from 130° to 145°. The efficiency droop was reduced from 46% to 35%, effects which can be attributed to the elimination of TDs and strain relaxation by embedded nanopatterns. In addition, the light output power of HP-LED at 360-mA injection currents exhibits a ∼ 22.3% enhancement, demonstrating that hybrid-PSSs are beneficial to apply in high-power LEDs.

  8. Towards an integrated optofluidic system for highly sensitive detection of antibiotics in seawater incorporating bimodal waveguide photonic biosensors and complex, active microfluidics

    NASA Astrophysics Data System (ADS)

    Szydzik, C.; Gavela, A. F.; Roccisano, J.; Herranz de Andrés, S.; Mitchell, A.; Lechuga, L. M.

    2016-12-01

    We present recent results on the realisation and demonstration of an integrated optofluidic lab-on-a-chip measurement system. The system consists of an integrated on-chip automated microfluidic fluid handling subsystem, coupled with bimodal nano-interferometer waveguide technology, and is applied in the context of detection of antibiotics in seawater. The bimodal waveguide (BMWG) is a highly sensitive label-free biosensor. Integration of complex microfluidic systems with bimodal waveguide technology enables on-chip sample handling and fluid processing capabilities and allows for significant automation of experimental processes. The on-chip fluid-handling subsystem is realised through the integration of pneumatically actuated elastomer pumps and valves, enabling high temporal resolution sample and reagent delivery and facilitating multiplexed detection processes.

  9. Actuation of digital micro drops by electrowetting on open microfluidic chips fabricated in photolithography.

    PubMed

    Ko, Hyojin; Lee, Jeong Soo; Jung, Chan-Hee; Choi, Jae-Hak; Kwon, Oh-Sun; Shin, Kwanwoo

    2014-08-01

    Basic manipulations of discrete liquid drops on opened microfluidic chips based on electrowetting on dielectrics were described. While most developed microfluidic chips are closed systems equipped with a top plate to cover mechanically and to contact electrically to drop samples, our chips are opened systems with a single plate without any electric contact to drops directly. The chips consist of a linear array of patterned electrodes at 1.8 mm pitch was fabricated on a glass plate coated with thin hydrophobic and dielectric layers by using various methods including photolithography, spin coating and ion sputtering. Several actuations such as lateral oscillation, colliding mergence and translational motion for 3-10 μL water drops have been demonstrated satisfactory. All these kinetic performances of opened chips were similar to those of closed chip systems, indicating superiority of a none-contact method for the transport of drops on opened microfluidic chips actuated by using electrowetting technique.

  10. Numerical simulation of isolation of cancer cells in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Djukic, T.; Topalovic, M.; Filipovic, N.

    2015-08-01

    Cancer is a disease that is characterized by the uncontrolled increase of numbers of cells. Circulating tumour cells (CTCs) are separated from the primary tumor, circulate in the bloodstream and form metastases. Circulating tumor cells can be identified in the blood of a patient by taking a blood sample. Microfluidic chips are a new technique that is used to isolate these cells from the blood sample. In this paper a numerical model is presented that is able to simulate the motion of individual cells through a microfluidic chip. The proposed numerical model gives very valuable insight into the processes happening within a microfluidic chip. The accuracy of the proposed model is compared with experimental results. The experimental setup that is described in literature is used to create identical geometrical domains and define simulation parameters. A good agreement of experimental and numerical results demonstrates that the proposed model can be successfully used to simulate complex behaviour of CTCs inside microfluidic chips.

  11. Flexible packaging of solid-state integrated circuit chips with elastomeric microfluidics

    PubMed Central

    Zhang, Bowei; Dong, Quan; Korman, Can E.; Li, Zhenyu; Zaghloul, Mona E.

    2013-01-01

    A flexible technology is proposed to integrate smart electronics and microfluidics all embedded in an elastomer package. The microfluidic channels are used to deliver both liquid samples and liquid metals to the integrated circuits (ICs). The liquid metals are used to realize electrical interconnects to the IC chip. This avoids the traditional IC packaging challenges, such as wire-bonding and flip-chip bonding, which are not compatible with current microfluidic technologies. As a demonstration we integrated a CMOS magnetic sensor chip and associate microfluidic channels on a polydimethylsiloxane (PDMS) substrate that allows precise delivery of small liquid samples to the sensor. Furthermore, the packaged system is fully functional under bending curvature radius of one centimetre and uniaxial strain of 15%. The flexible integration of solid-state ICs with microfluidics enables compact flexible electronic and lab-on-a-chip systems, which hold great potential for wearable health monitoring, point-of-care diagnostics and environmental sensing among many other applications.

  12. Chip-based wide field-of-view nanoscopy

    NASA Astrophysics Data System (ADS)

    Diekmann, Robin; Helle, Øystein I.; Øie, Cristina I.; McCourt, Peter; Huser, Thomas R.; Schüttpelz, Mark; Ahluwalia, Balpreet S.

    2017-04-01

    Present optical nanoscopy techniques use a complex microscope for imaging and a simple glass slide to hold the sample. Here, we demonstrate the inverse: the use of a complex, but mass-producible optical chip, which hosts the sample and provides a waveguide for the illumination source, and a standard low-cost microscope to acquire super-resolved images via two different approaches. Waveguides composed of a material with high refractive-index contrast provide a strong evanescent field that is used for single-molecule switching and fluorescence excitation, thus enabling chip-based single-molecule localization microscopy. Additionally, multimode interference patterns induce spatial fluorescence intensity variations that enable fluctuation-based super-resolution imaging. As chip-based nanoscopy separates the illumination and detection light paths, total-internal-reflection fluorescence excitation is possible over a large field of view, with up to 0.5 mm × 0.5 mm being demonstrated. Using multicolour chip-based nanoscopy, we visualize fenestrations in liver sinusoidal endothelial cells.

  13. Single chip camera device having double sampling operation

    NASA Technical Reports Server (NTRS)

    Fossum, Eric R. (Inventor); Nixon, Robert (Inventor)

    2002-01-01

    A single chip camera device is formed on a single substrate including an image acquisition portion for control portion and the timing circuit formed on the substrate. The timing circuit also controls the photoreceptors in a double sampling mode in which are reset level is first read and then after an integration time a charged level is read.

  14. Chip-to-chip optical link by using optical wiring method

    NASA Astrophysics Data System (ADS)

    Cho, In-Kui; Ahn, Seoung Ho; Jeong, Myung-Yung; Rho, Byung Sup; Park, Hyo Hoon

    2008-01-01

    A practical optical link system was prepared with a transmitter (Tx) and receiver (Rx). The optical TRx module consisted of a metal optical bench, a module printed circuit board (PCB), a driver/receiver IC, a VCSEL/PD array, and an optical link block composed of plastic optical fiber (POF). For the optical interconnection between the light-sources and detectors, an optical wiring method has been proposed to enable easy assembly. This paper provides a method for optical interconnection between an optical Tx and an optical Rx, comprising the following steps: (a) forming a light source device, an optical detection device, and an optical transmission unit on a substrate (metal optical bench (MOB)); (b) preparing a flexible optical transmission-connection medium (optical wiring link) to optically connect the light source device formed on the substrate with the optical detection device; and (c) directly connecting one end of the surface-finished optical transmission connection medium with the light source device and the other end with the optical detection device. A chip-to-chip optical link system constructed with TRx modules was fabricated and the optical characteristics were measured. The results clearly demonstrate that the use of an optical wiring method can provide robust and cost-effective assembly for vertical-cavity surface-emitting lasers (VCSELs) and photodiodes (PDs). We successfully achieved a 5 Gb/s data transmission rate with this optical link.

  15. Thermal processing of bone: in vitro response of mesenchymal cells to bone-conditioned medium.

    PubMed

    Sawada, K; Caballé-Serrano, J; Schuldt Filho, G; Bosshardt, D D; Schaller, B; Buser, D; Gruber, R

    2015-08-01

    The autoclaving, pasteurization, and freezing of bone grafts to remove bacteria and viruses, and for preservation, respectively, is considered to alter biological properties during graft consolidation. Fresh bone grafts release paracrine-like signals that are considered to support tissue regeneration. However, the impact of the autoclaving, pasteurization, and freezing of bone grafts on paracrine signals remains unknown. Therefore, conditioned medium was prepared from porcine cortical bone chips that had undergone thermal processing. The biological properties of the bone-conditioned medium were assessed by examining the changes in expression of target genes in oral fibroblasts. The data showed that conditioned medium obtained from bone chips that had undergone pasteurization and freezing changed the expression of adrenomedullin, pentraxin 3, BTB/POZ domain-containing protein 11, interleukin 11, NADPH oxidase 4, and proteoglycan 4 by at least five-fold in oral fibroblasts. Bone-conditioned medium obtained from autoclaved bone chips, however, failed to change the expression of the respective genes. Also, when bone-conditioned medium was prepared from fresh bone chips, autoclaving blocked the capacity of bone-conditioned medium to modulate gene expression. These in vitro results suggest that pasteurization and freezing of bone grafts preserve the release of biologically active paracrine signals, but autoclaving does not. Copyright © 2015 International Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  16. Nanoporous Glass Integrated in Volumetric Bar-Chart Chip for Point-of-Care Diagnostics of Non-Small Cell Lung Cancer.

    PubMed

    Li, Ying; Xuan, Jie; Song, Yujun; Qi, Wenjin; He, Bangshun; Wang, Ping; Qin, Lidong

    2016-01-26

    Point-of-care (POC) testing has the potential to enable rapid, low-cost, and large-scale screening. POC detection of a multiplexed biomarker panel can facilitate the early diagnosis of non-small cell lung cancer (NSCLC) and, thus, may allow for more timely surgical intervention for life-saving treatment. Herein, we report the nanoporous glass (NPG) integrated volumetric bar-chart chip (V-Chip) for POC detection of the three NSCLC biomarkers CEA, CYFRA 21-1, and SCCA, by the naked eye. The 3D nanostructures in the NPG membrane efficiently increase the number of binding sites for antibodies and decrease the diffusion distance between antibody and antigen, enabling the low detection limit and rapid analysis time of the NPG-V-Chip. We utilized the NPG-V-Chip to test the NSCLC biomarker panel and found that the limit of detection can reach 50 pg/mL (10-fold improvement over the original V-Chip), and the total assay time can be decreased from 4 to 0.5 h. We then detected CEA in 21 serum samples from patients with common cancers, and the on-chip results showed good correlation with the clinical results. We further assayed 10 lung cancer samples using the device and confirmed the results obtained using conventional ELISA methods. In summary, the NPG-V-Chip platform has the ability of multiplex, low detection limit, low cost, lack of need for accessory equipment, and rapid analysis time, which may render the V-Chip a useful platform for quantitative POC detection in resource-limited settings and personalized diagnostics.

  17. Flip-chip assembly and reliability using gold/tin solder bumps

    NASA Astrophysics Data System (ADS)

    Oppermann, Hermann; Hutter, Matthias; Klein, Matthias; Reichl, Herbert

    2004-09-01

    Au/Sn solder bumps are commonly used for flip chip assembly of optoelectronic and RF devices. They allow a fluxless assembly which is required to avoid contamination at optical interfaces. Flip chip assembly experiments were carried out using as plated Au/Sn bumps without prior bump reflow. An RF and reliability test vehicles comprise a GaAs chip which was flip chip soldered on a silicon substrate. Temperature cycling tests with and without underfiller were performed and the results are presented. The different failure modes for underfilled and non-underfilled samples were discussed and compared. Additional reliability tests were performed with flip chip bonding by gold thermocompression for comparison. The test results and the failure modes are discussed in detail.

  18. HPLC-DAD method development and validation for the quantification of hydroxymethylfurfural in corn chips by means of response surface optimisation.

    PubMed

    Salvatierra Virgen, Sara; Ceballos-Magaña, Silvia Guillermina; Salvatierra-Stamp, Vilma Del Carmen; Sumaya-Martínez, Maria Teresa; Martínez-Martínez, Francisco Javier; Muñiz-Valencia, Roberto

    2017-12-01

    In recent years, there has been an increased concern about the presence of toxic compounds derived from the Maillard reaction produced during food cooking at high temperatures. The main toxic compounds derived from this reaction are acrylamide and hydroxymethylfurfural (HMF). The majority of analytical methods require sample treatments using solvents which are highly polluting for the environment. The difficulty of quantifying HMF in complex fried food matrices encourages the development of new analytical methods. This paper provides a rapid, sensitive and environmentally-friendly analytical method for the quantification of HMF in corn chips using HPLC-DAD. Chromatographic separation resulted in a baseline separation for HMF in 3.7 min. Sample treatment for corn chip samples first involved a leaching process using water and afterwards a solid-phase extraction (SPE) using HLB-Oasis polymeric cartridges. Sample treatment optimisation was carried out by means of Box-Behnken fractional factorial design and Response Surface Methodolog y to examine the effects of four variables (sample weight, pH, sonication time and elution volume) on HMF extraction from corn chips. The SPE-HPLC-DAD method was validated. The limits of detection and quantification were 0.82 and 2.20 mg kg -1 , respectively. Method precision was evaluated in terms of repeatability and reproducibility as relative standard deviations (RSDs) using three concentration levels. For repeatability, RSD values were 6.9, 3.6 and 2.0%; and for reproducibility 18.8, 7.9 and 2.9%. For a ruggedness study the Yuden test was applied and the result demonstrated the method as robust. The method was successfully applied to different corn chip samples.

  19. On-chip clearing of arrays of 3-D cell cultures and micro-tissues.

    PubMed

    Grist, S M; Nasseri, S S; Poon, T; Roskelley, C; Cheung, K C

    2016-07-01

    Three-dimensional (3-D) cell cultures are beneficial models for mimicking the complexities of in vivo tissues, especially in tumour studies where transport limitations can complicate response to cancer drugs. 3-D optical microscopy techniques are less involved than traditional embedding and sectioning, but are impeded by optical scattering properties of the tissues. Confocal and even two-photon microscopy limit sample imaging to approximately 100-200 μm depth, which is insufficient to image hypoxic spheroid cores. Optical clearing methods have permitted high-depth imaging of tissues without physical sectioning, but they are difficult to implement for smaller 3-D cultures due to sample loss in solution exchange. In this work, we demonstrate a microfluidic platform for high-throughput on-chip optical clearing of breast cancer spheroids using the SeeDB, Clear(T2), and ScaleSQ clearing methods. Although all three methods are able to effectively clear the spheroids, we find that SeeDB and ScaleSQ more effectively clear the sample than Clear(T2); however, SeeDB induces green autofluorescence while ScaleS causes sample expansion. Our unique on-chip implementation permits clearing arrays of 3-D cultures using perfusion while monitoring the 3-D cultures throughout the process, enabling visualization of the clearing endpoint as well as monitoring of transient changes that could induce image artefacts. Our microfluidic device is compatible with on-chip 3-D cell culture, permitting the use of on-chip clearing at the endpoint after monitoring the same spheroids during their culture. This on-chip method has the potential to improve readout from 3-D cultures, facilitating their use in cell-based assays for high-content drug screening and other applications.

  20. A 1-channel 3-band wide dynamic range compression chip for vibration transducer of implantable hearing aids.

    PubMed

    Kim, Dongwook; Seong, Kiwoong; Kim, Myoungnam; Cho, Jinho; Lee, Jyunghyun

    2014-01-01

    In this paper, a digital audio processing chip which uses a wide dynamic range compression (WDRC) algorithm is designed and implemented for implantable hearing aids system. The designed chip operates at a single voltage of 3.3V and drives a 16 bit parallel input and output at 32 kHz sample. The designed chip has 1-channel 3-band WDRC composed of a FIR filter bank, a level detector, and a compression part. To verify the performance of the designed chip, we measured the frequency separations of bands and compression gain control to reflect the hearing threshold level.

  1. Hybridization of Environmental Microbial Community Nucleic Acids by GeoChip.

    PubMed

    Van Nostrand, Joy D; Yin, Huaqin; Wu, Liyou; Yuan, Tong; Zhou, Jizhong

    2016-01-01

    Functional gene arrays, like the GeoChip, allow for the study of tens of thousands of genes in a single assay. The GeoChip array (5.0) contains probes for genes involved in geochemical cycling (N, C, S, and P), metal homeostasis, stress response, organic contaminant degradation, antibiotic resistance, secondary metabolism, and virulence factors as well as genes specific for fungi, protists, and viruses. Here, we briefly describe GeoChip design strategies (gene selection and probe design) and discuss minimum quantity and quality requirements for nucleic acids. We then provide detailed protocols for amplification, labeling, and hybridization of samples to the GeoChip.

  2. Tyrosine sulfation in N-terminal domain of human C5a receptor is necessary for binding of chemotaxis inhibitory protein of Staphylococcus aureus

    PubMed Central

    Liu, Zhen-jia; Yang, Yan-juan; Jiang, Lei; Xu, Ying-chun; Wang, Ai-xia; Du, Guan-hua; Gao, Jin-ming

    2011-01-01

    Aim: Staphylococcus aureus evades host defense through releasing several virulence proteins, such as chemotaxis inhibitory protein of staphylococcus aureus (CHIPS). It has been shown that extracellular N terminus of C5a receptor (C5aR) forms the binding domain for CHIPS, and tyrosine sulfation is emerging as a key factor in determining protein-protein interaction. The aim of this study was to evaluate the role of tyrosine sulfation of N-terminal of C5aR in its binding with CHIPS. Methods: Expression plasmids encoding C5aR and its mutants were prepared using PCR and site-directed mutagenesis and were used to transfect HEK 293T cells using calcium phosphate. Recombinant CHIPS protein was purified. Western blotting was used to examine the binding efficiency of CHIPS to C5aR or its mutants. Results: CHIPS exclusively binds to C5aR, but not to C5L2 or C3aR. A nonspecific sulfation inhibitor, sodium chlorate (50 nmol/L), diminishes the binding ability of C5aR with CHIPS. Blocking sulfation by mutation of tyrosine to phenylalanine at positions 11 and 14 of C5aR N terminus, which blocked sulfation, completely abrogates CHIPS binding. When tyrosine 14 alone was mutated to phenylalanine, the binding efficiency of recombinant CHIPS was substantially decreased. Conclusion: The results demonstrate a structural basis of C5aR-CHIPS association, in which tyrosine sulfation of N-terminal C5aR plays an important role. Our data may have potential significance in development of novel drugs for therapeutic intervention. PMID:21706042

  3. Cassava chip (Manihot esculenta Crantz) as an energy source for ruminant feeding.

    PubMed

    Wanapat, Metha; Kang, Sungchhang

    2015-12-01

    Cassava ( Manihot esculenta Crantz) is widely grown in sub-tropical and tropical areas, producing roots as an energy source while the top biomass including leaves and immature stems can be sun-dried and used as cassava hay. Cassava roots can be processed as dried chip or pellet. It is rich in soluble carbohydrate (75 to 85%) but low in crude protein (2 to 3%). Its energy value is comparable to corn meal but has a relatively higher rate of rumen degradation. Higher levels of non-protein nitrogen especially urea (1 to 4%) can be successfully incorporated in concentrates containing cassava chip as an energy source. Cassava chip can also be processed with urea and other ingredients (tallow, sulfur, raw banana meal, cassava hay, and soybean meal) to make products such as cassarea, cassa-ban, and cassaya. Various studies have been conducted in ruminants using cassava chip to replace corn meal in the concentrate mixtures and have revealed satisfactory results in rumen fermentation efficiency and the subsequent production of meat and milk. In addition, it was advantageous when used in combination with rice bran in the concentrate supplement. Practical home-made-concentrate using cassava chip can be easily prepared for use on farms. A recent development has involved enriching protein in cassava chips, yielding yeast fermented cassava chip protein (YEFECAP) of up to 47.5% crude protein, which can be used to replace soybean meal. It is therefore, recommended to use cassava chip as an alternative source of energy to corn meal when the price is economical and it is locally available.

  4. Experiences with Lab-on-a-chip Technology in Support of NASA Supported Research

    NASA Technical Reports Server (NTRS)

    Monaco, Lisa

    2003-01-01

    Under the auspices of the Microgravity Sciences and Application Department at Marshall Space Flight Center, we have custom designed and fabricated a lab-on-a-chip (LOC) device, along with Caliper Technologies, for macromolecular crystal growth. The chip has been designed to deliver specified proportions of up-to five various constituents to one of two growth wells (on-chip) for crystal growth. To date, we have grown crystals of thaumatin, glucose isomerase and appoferitin on the chip. The LOC approach offered many advantages that rendered it highly suitable for space based hardware to perform crystal growth on the International Space Station. The same hardware that was utilized for the crystal growth investigations, has also been used by researchers at Glenn Research Center to investigate aspects of microfluidic phenomenon associated with two-phase flow. Additionally, our LOCAD (Lab-on-a-chip Application Development) team has lent its support to Johnson Space Center s Modular Assay for Solar System Exploration project. At present, the LOCAD team is working on the design and build of a unique lab-on-a-chip breadboard control unit whose function is not commercially available. The breadboard can be used as a test bed for the development of chip size labs for environmental monitoring, crew health monitoring assays, extended flight pharmacological preparations, and many more areas. This unique control unit will be configured for local use and/or remote operation, via the Internet, by other NASA centers. The lab-on-a-chip control unit is being developed with the primary goal of meeting Agency level strategic goals.

  5. Gold-coated polydimethylsiloxane microwells for high-throughput electrochemiluminescence analysis of intracellular glucose at single cells.

    PubMed

    Xia, Juan; Zhou, Junyu; Zhang, Ronggui; Jiang, Dechen; Jiang, Depeng

    2018-06-04

    In this communication, a gold-coated polydimethylsiloxane (PDMS) chip with cell-sized microwells was prepared through a stamping and spraying process that was applied directly for high-throughput electrochemiluminescence (ECL) analysis of intracellular glucose at single cells. As compared with the previous multiple-step fabrication of photoresist-based microwells on the electrode, the preparation process is simple and offers fresh electrode surface for higher luminescence intensity. More luminescence intensity was recorded from cell-retained microwells than that at the planar region among the microwells that was correlated with the content of intracellular glucose. The successful monitoring of intracellular glucose at single cells using this PDMS chip will provide an alternative strategy for high-throughput single-cell analysis. Graphical abstract ᅟ.

  6. Development and evaluation of a real-time fluorogenic loop-mediated isothermal amplification assay integrated on a microfluidic disc chip (on-chip LAMP) for rapid and simultaneous detection of ten pathogenic bacteria in aquatic animals.

    PubMed

    Zhou, Qian-Jin; Wang, Lei; Chen, Jiong; Wang, Rui-Na; Shi, Yu-Hong; Li, Chang-Hong; Zhang, De-Min; Yan, Xiao-Jun; Zhang, Yan-Jun

    2014-09-01

    Rapid, low-cost, and user-friendly strategies are urgently needed for early disease diagnosis and timely treatment, particularly for on-site screening of pathogens in aquaculture. In this study, we successfully developed a real-time fluorogenic loop-mediated isothermal amplification assay integrated on a microfluidic disc chip (on-chip LAMP), which was capable of simultaneously detecting 10 pathogenic bacteria in aquatic animals, i.e., Nocardia seriolae, Pseudomonas putida, Streptococcus iniae, Vibrio alginolyticus, Vibrio anguillarum, Vibrio fluvialis, Vibrio harveyi, Vibrio parahaemolyticus, Vibrio rotiferianus, and Vibrio vulnificus. The assay provided a nearly-automated approach, with only a single pipetting step per chip for sample dispensing. This technique could achieve limits of detection (LOD) ranging from 0.40 to 6.42pg per 1.414μL reaction in less than 30 min. The robust reproducibility was demonstrated by a little variation among duplications for each bacterium with the coefficient of variation (CV) for time to positive (Tp) value less than 0.10. The clinical sensitivity and specificity of this on-chip LAMP assay in detecting field samples were 96.2% and 93.8% by comparison with conventional microbiological methods. Compared with other well-known techniques, on-chip LAMP assay provides low sample and reagent consumption, ease-of-use, accelerated analysis, multiple bacteria and on-site detection, and high reproducibility, indicating that such a technique would be applicable for on-site detection and routine monitoring of multiple pathogens in aquaculture. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Selective isolation of magnetic nanoparticle-mediated heterogeneity subpopulation of circulating tumor cells using magnetic gradient based microfluidic system.

    PubMed

    Kwak, Bongseop; Lee, Jaehun; Lee, Dongkyu; Lee, Kangho; Kwon, Ohwon; Kang, Shinwon; Kim, Youngwoo

    2017-02-15

    Relocation mechanisms of the circulating tumor cells (CTCs) from the primary site to the secondary site through the blood vessel network cause tumor metastasis. Despite of the importance to diagnose the cancer metastasis by CTCs, still it is formidable challenge to use in the clinical purpose because of the rarity and the heterogeneity of CTCs in the cancer patient's peripheral blood sample. In this study we have developed magnetic force gradient based microfluidic chip (Mag-Gradient Chip) for isolating the total number of CTCs in the sample and characterizing the state of CTCs simultaneously with respect to the epithelial cell adhesion molecule (EpCAM) expression level. We have synthesized magnetic nanoparticles (MNPs) using hydrothermal method and functionalized anti-EpCAM on their surface for the specific binding with CTCs. The Mag-Gradient Chip designed to isolate and classify the CTCs by isolating at the different location in the chip using magnetic force differences depending on the EpCAM expression level. We observed 95.7% of EpCAM positive and 79.3% of EpCAM negative CTCs isolated in the Mag-Gradient Chip. At the same time, the 71.3% of isolated EpCAM positive CTCs were isolated at the first half area whereas the 76.9% of EpCAM negative CTCs were collected at the latter half area. The Mag-Gradient Chip can isolate the 3ml of heterogeneous CTCs sample in 1h with high isolating yield. The EpCAM expression level dose not means essential condition of the metastatic CTCs, but the Mag-Gradient Chip can shorten the date to diagnose the cancer metastasis in clinic. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Studies on spectroscopy of glycerol in THz range using microfluidic chip-integrated micropump

    NASA Astrophysics Data System (ADS)

    Su, Bo; Han, Xue; Wu, Ying; Zhang, Cunlin

    2014-11-01

    Terahertz time-domain spectroscopy (THz-TDS) is a detection method of biological molecules with label-free, non-ionizing, non-intrusive, no pollution and real-time monitoring. But owing to the strong THz absorption by water, it is mainly used in the solid state detection of biological molecules. In this paper, we present a microfluidic chip technique for detecting biological liquid samples using the transmission type of THz-TDS system. The microfluidic channel of the microfluidic chip is fabricated in the quartz glass using Micro-Electro-Mechanical System (MEMS) technology and sealed with polydimethylsiloxane (PDMS) diaphragm. The length, width and depth of the microfluidic channel are 25mm, 100μm and 50μm, respectively. The diameter of THz detection zone in the microfluidic channel is 4mm. The thicknesses of quartz glass and PDMS diaphragm are 1mm and 250μm, individually. Another one of the same quartz glass is used to bond with the PDMS for the rigidity and air tightness of the microfluidic chip. In order to realize the automation of sampling and improve the control precise of fluid, a micropump, which comprises PDMS diaphragm, pump chamber, diffuser and nozzle and flat vibration motor, is integrated on the microfluidic chip. The diffuser and nozzle are fabricated on both sides of the pump chamber, which is covered with PDMS diaphragm. The flat vibration motor is stuck on the PDMS diaphragm as the actuator. We study the terahertz absorption spectroscopy characteristics of glycerol with the concentration of 98% in the microfluidic chip by the aid of the THz-TDS system, and the feasibility of the microfluidic chip for the detection of liquid samples is proved.

  9. [Development of molecular detection of food-borne pathogenic bacteria using miniaturized microfluidic devices].

    PubMed

    Iván, Kristóf; Maráz, Anna

    2015-12-20

    Detection and identification of food-borne pathogenic bacteria are key points for the assurance of microbiological food safety. Traditional culture-based methods are more and more replaced by or supplemented with nucleic acid based molecular techniques, targeting specific (preferably virulence) genes in the genomes. Internationally validated DNA amplification - most frequently real-time polymerase chain reaction - methods are applied by the food microbiological testing laboratories for routine analysis, which will result not only in shortening the time for results but they also improve the performance characteristics (e.g. sensitivity, specificity) of the methods. Beside numerous advantages of the polymerase chain reaction based techniques for routine microbiological analysis certain drawbacks have to be mentioned, such as the high cost of the equipment and reagents, as well as the risk of contamination of the laboratory environment by the polymerase chain reaction amplicons, which require construction of an isolated laboratory system. Lab-on-a-chip systems can integrate most of these laboratory processes within a miniaturized device that delivers the same specificity and reliability as the standard protocols. The benefits of miniaturized devices are: simple - often automated - use, small overall size, portability, sterility due to single use possibility. These miniaturized rapid diagnostic tests are being researched and developed at the best research centers around the globe implementing various sample preparation and molecular DNA amplification methods on-chip. In parallel, the aim of the authors' research is to develop microfluidic Lab-on-a-chip devices for the detection and identification of food-borne pathogenic bacteria.

  10. An accessible micro-capillary electrophoresis device using surface-tension-driven flow

    PubMed Central

    Mohanty, Swomitra K.; Warrick, Jay; Gorski, Jack; Beebe, David J.

    2010-01-01

    We present a rapidly fabricated micro-capillary electrophoresis chip that utilizes surface-tension-driven flow for sample injection and extraction of DNA. Surface-tension-driven flow (i.e. passive pumping) injects a fixed volume of sample that can be predicted mathematically. Passive pumping eliminates the need for tubing, valves, syringe pumps, and other equipment typically needed for interfacing with microelectrophoresis chips. This method requires a standard micropipette to load samples before separation, and remove the resulting bands after analysis. The device was made using liquid phase photopolymerization to rapidly fabricate the chip without the need of special equipment typically associated with the construction of microelectrophoresis chips (e.g. cleanroom). Batch fabrication time for the device presented here was 1.5 h including channel coating time to suppress electroosmotic flow. Devices were constructed out of poly-isobornyl acrylate and glass. A standard microscope with a UV source was used for sample detection. Separations were demonstrated using Promega BenchTop 100 bp ladder in hydroxyl ethyl cellulose (HEC) and oligonucleotides of 91 and 118 bp were used to characterize sample injection and extraction of DNA bands. The end result was an inexpensive micro-capillary electrophoresis device that uses tools (e.g. micropipette, electrophoretic power supplies, and microscopes) already present in most labs for sample manipulation and detection, making it more accessible for potential end users. PMID:19425002

  11. Reusable and Mediator-Free Cholesterol Biosensor Based on Cholesterol Oxidase Immobilized onto TGA-SAM Modified Smart Bio-Chips

    PubMed Central

    Rahman, Mohammed M.

    2014-01-01

    A reusable and mediator-free cholesterol biosensor based on cholesterol oxidase (ChOx) was fabricated based on self-assembled monolayer (SAM) of thioglycolic acid (TGA) (covalent enzyme immobilization by dropping method) using bio-chips. Cholesterol was detected with modified bio-chip (Gold/Thioglycolic-acid/Cholesterol-oxidase i.e., Au/TGA/ChOx) by reliable cyclic voltammetric (CV) technique at room conditions. The Au/TGA/ChOx modified bio-chip sensor demonstrates good linearity (1.0 nM to 1.0 mM; R = 0.9935), low-detection limit (∼0.42 nM, SNR∼3), and higher sensitivity (∼74.3 µAµM−1cm−2), lowest-small sample volume (50.0 μL), good stability, and reproducibility. To the best of our knowledge, this is the first statement with a very high sensitivity, low-detection limit, and low-sample volumes are required for cholesterol biosensor using Au/TGA/ChOx-chips assembly. The result of this facile approach was investigated for the biomedical applications for real samples at room conditions with significant assembly (Au/TGA/ChOx) towards the development of selected cholesterol biosensors, which can offer analytical access to a large group of enzymes for wide range of biomedical applications in health-care fields. PMID:24949733

  12. Overview of Considerations in Assessing the Biomass Potential of Army Installations.

    DTIC Science & Technology

    1981-08-01

    stage. Will the species grow well in poor soils and on harsh, open sites? Trees that met these standards were then grouped according to their...frequency of fire, (2) reduces the need of fire control methods such as controlled burns, and (3) makes site preparation easier.21 Whole-tree chipping...the "aesthetic" value of the stand is increased.22 The negative effect most often thought to occur with whole-tree chipping is loss of soil nutrients

  13. First all-in-one diagnostic tool for DNA intelligence: genome-wide inference of biogeographic ancestry, appearance, relatedness, and sex with the Identitas v1 Forensic Chip.

    PubMed

    Keating, Brendan; Bansal, Aruna T; Walsh, Susan; Millman, Jonathan; Newman, Jonathan; Kidd, Kenneth; Budowle, Bruce; Eisenberg, Arthur; Donfack, Joseph; Gasparini, Paolo; Budimlija, Zoran; Henders, Anjali K; Chandrupatla, Hareesh; Duffy, David L; Gordon, Scott D; Hysi, Pirro; Liu, Fan; Medland, Sarah E; Rubin, Laurence; Martin, Nicholas G; Spector, Timothy D; Kayser, Manfred

    2013-05-01

    When a forensic DNA sample cannot be associated directly with a previously genotyped reference sample by standard short tandem repeat profiling, the investigation required for identifying perpetrators, victims, or missing persons can be both costly and time consuming. Here, we describe the outcome of a collaborative study using the Identitas Version 1 (v1) Forensic Chip, the first commercially available all-in-one tool dedicated to the concept of developing intelligence leads based on DNA. The chip allows parallel interrogation of 201,173 genome-wide autosomal, X-chromosomal, Y-chromosomal, and mitochondrial single nucleotide polymorphisms for inference of biogeographic ancestry, appearance, relatedness, and sex. The first assessment of the chip's performance was carried out on 3,196 blinded DNA samples of varying quantities and qualities, covering a wide range of biogeographic origin and eye/hair coloration as well as variation in relatedness and sex. Overall, 95 % of the samples (N = 3,034) passed quality checks with an overall genotype call rate >90 % on variable numbers of available recorded trait information. Predictions of sex, direct match, and first to third degree relatedness were highly accurate. Chip-based predictions of biparental continental ancestry were on average ~94 % correct (further support provided by separately inferred patrilineal and matrilineal ancestry). Predictions of eye color were 85 % correct for brown and 70 % correct for blue eyes, and predictions of hair color were 72 % for brown, 63 % for blond, 58 % for black, and 48 % for red hair. From the 5 % of samples (N = 162) with <90 % call rate, 56 % yielded correct continental ancestry predictions while 7 % yielded sufficient genotypes to allow hair and eye color prediction. Our results demonstrate that the Identitas v1 Forensic Chip holds great promise for a wide range of applications including criminal investigations, missing person investigations, and for national security purposes.

  14. Microfluidic-Based Robotic Sampling System for Radioactive Solutions

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jack D. Law; Julia L. Tripp; Tara E. Smith

    A novel microfluidic based robotic sampling system has been developed for sampling and analysis of liquid solutions in nuclear processes. This system couples the use of a microfluidic sample chip with a robotic system designed to allow remote, automated sampling of process solutions in-cell and facilitates direct coupling of the microfluidic sample chip with analytical instrumentation. This system provides the capability for near real time analysis, reduces analytical waste, and minimizes the potential for personnel exposure associated with traditional sampling methods. A prototype sampling system was designed, built and tested. System testing demonstrated operability of the microfluidic based sample systemmore » and identified system modifications to optimize performance.« less

  15. Solution-based circuits enable rapid and multiplexed pathogen detection.

    PubMed

    Lam, Brian; Das, Jagotamoy; Holmes, Richard D; Live, Ludovic; Sage, Andrew; Sargent, Edward H; Kelley, Shana O

    2013-01-01

    Electronic readout of markers of disease provides compelling simplicity, sensitivity and specificity in the detection of small panels of biomarkers in clinical samples; however, the most important emerging tests for disease, such as infectious disease speciation and antibiotic-resistance profiling, will need to interrogate samples for many dozens of biomarkers. Electronic readout of large panels of markers has been hampered by the difficulty of addressing large arrays of electrode-based sensors on inexpensive platforms. Here we report a new concept--solution-based circuits formed on chip--that makes highly multiplexed electrochemical sensing feasible on passive chips. The solution-based circuits switch the information-carrying signal readout channels and eliminate all measurable crosstalk from adjacent, biomolecule-specific microsensors. We build chips that feature this advance and prove that they analyse unpurified samples successfully, and accurately classify pathogens at clinically relevant concentrations. We also show that signature molecules can be accurately read 2  minutes after sample introduction.

  16. 3D on-chip microscopy of optically cleared tissue

    NASA Astrophysics Data System (ADS)

    Zhang, Yibo; Shin, Yoonjung; Sung, Kevin; Yang, Sam; Chen, Harrison; Wang, Hongda; Teng, Da; Rivenson, Yair; Kulkarni, Rajan P.; Ozcan, Aydogan

    2018-02-01

    Traditional pathology relies on tissue biopsy, micro-sectioning, immunohistochemistry and microscopic imaging, which are relatively expensive and labor-intensive, and therefore are less accessible in resource-limited areas. Low-cost tissue clearing techniques, such as the simplified CLARITY method (SCM), are promising to potentially reduce the cost of disease diagnosis by providing 3D imaging and phenotyping of thicker tissue samples with simpler preparation steps. However, the mainstream imaging approach for cleared tissue, fluorescence microscopy, suffers from high-cost, photobleaching and signal fading. As an alternative approach to fluorescence, here we demonstrate 3D imaging of SCMcleared tissue using on-chip holography, which is based on pixel-super-resolution and multi-height phase recovery algorithms to digitally compute the sample's amplitude and phase images at various z-slices/depths through the sample. The tissue clearing procedures and the lens-free imaging system were jointly optimized to find the best illumination wavelength, tissue thickness, staining solution pH, and the number of hologram heights to maximize the imaged tissue volume, minimize the amount of acquired data, while maintaining a high contrast-to-noise ratio for the imaged cells. After this optimization, we achieved 3D imaging of a 200-μm thick cleared mouse brain tissue over a field-of-view of <20mm2 , and the resulting 3D z-stack agrees well with the images acquired with a scanning lens-based microscope (20× 0.75NA). Moreover, the lens-free microscope achieves an order-of-magnitude better data efficiency compared to its lens-based counterparts for volumetric imaging of samples. The presented low-cost and high-throughput lens-free tissue imaging technique enabled by CLARITY can be used in various biomedical applications in low-resource-settings.

  17. A portable x-ray fluorescence instrument for analyzing dust wipe samples for lead: evaluation with field samples.

    PubMed

    Sterling, D A; Lewis, R D; Luke, D A; Shadel, B N

    2000-06-01

    Dust wipe samples collected in the field were tested by nondestructive X-ray fluorescence (XRF) followed by laboratory analysis with flame atomic absorption spectrophotometry (FAAS). Data were analyzed for precision and accuracy of measurement. Replicate samples with the XRF show high precision with an intraclass correlation coefficient (ICC) of 0.97 (P<0.0001) and an overall coefficient of variation of 11.6%. Paired comparison indicates no statistical difference (P=0.272) between XRF and FAAS analysis. Paired samples are highly correlated with an R(2) ranging between 0.89 for samples that contain paint chips and 0.93 for samples that do not contain paint chips. The ICC for absolute agreement between XRF and laboratory results was 0.95 (P<0.0001). The relative error over the concentration range of 25 to 14,200 microgram Pb is -12% (95% CI, -18 to -5). The XRF appears to be an excellent method for rapid on-site evaluation of dust wipes for clearance and risk assessment purposes, although there are indications of some confounding when paint chips are present. Copyright 2000 Academic Press.

  18. [A heart function measuring and analyzing instrument based on single-chip microcomputer].

    PubMed

    Rong, Z; Liang, H; Wang, S

    1999-05-01

    An Introduction a measuring and analyzing instrument, based on the single-chip microcomputer, which provides sample gathering, processing, controlling, adjusting, keyboard and printing. All informations are provided and displayed in Chinese.

  19. An electrochemical albumin-sensing system utilizing microfluidic technology

    NASA Astrophysics Data System (ADS)

    Huang, Chao-June; Lu, Chiu-Chun; Lin, Thong-Yueh; Chou, Tse-Chuan; Lee, Gwo-Bin

    2007-04-01

    This paper reports an integrated microfluidic chip capable of detecting the concentration of albumin in urine by using an electrochemical method in an automatic format. The integrated microfluidic chip was fabricated by using microelectromechanical system techniques. The albumin detection was conducted by using the electrochemical sensing method, in which the albumin in urine was detected by measuring the difference of peak currents between a bare reference electrode and an albumin-adsorption electrode. To perform the detection of the albumin in an automatic format, pneumatic microvalves and micropumps were integrated onto the microfluidic chip. The albumin sample and interference mixture solutions such as homovanillic acid, dopamine, norepinephrine and epinephrine were first stored in one of the three reservoirs. Then the solution comprising the albumin sample and interference solutions was transported to pass through the detection zone utilizing the pneumatic micropump. Experimental data showed that the developed system can successfully detect the concentration of the albumin in the existence of interference materials. When compared with the traditional albumin-sensing method, smaller amounts of samples were required to perform faster detection by using the integrated microfluidic chip. Additionally, the microfluidic chip integrated with pneumatic micropumps and microvalves facilitates the transportation of the samples in an automatic mode with lesser human intervention. The development of the integrated microfluidic albumin-sensing system may be promising for biomedical applications. Preliminary results of the current paper were presented at the 2nd International Meeting on Microsensors and Microsystems 2006 (National Cheng Kung University, Tainan, Taiwan, 15-18 January).

  20. Trace analysis of glyphosate in water by capillary electrophoresis on a chip with high sample volume loadability.

    PubMed

    Horčičiak, Michal; Masár, Marián; Bodor, Róbert; Danč, Ladislav; Bel, Peter

    2012-03-01

    A new method for the determination of trace glyphosate (GLYP), non-selective pesticide, by CZE with online ITP pre-treatment of drinking waters on a column-coupling (CC) chip has been developed. CC chip was equipped with two injection channels of 0.9 and 9.9 μL volumes, two separation channels of 9.3 μL total volume and a pair of conductivity detectors. A very effective ITP sample clean-up performed in the first channel at low pH (3.2) was introduced for quick CZE resolution and detection of GLYP carried out at higher pH (6.1) in the second channel on the CC chip. The LOD for GLYP was estimated at 2.5 μg/L (15 nmol/L) using a 9.9 |mL volume of the injection channel. ITP-CZE analyses of model and real samples have provided very favorable intra-day (0.1-1.2% RSD) and inter-day (2.9% RSD) repeatabilities of the migration time for GLYP while 0.2-6.9% RSD values were typical for the peak area data. Recoveries of GLYP in spiked drinking water varied in the range of 99-109%. A minimum pre-treatment of drinking water (degassing and dilution) and a short analysis time (ca. 10 min) were distinctive features of ITP-CZE determinations of GLYP on the CC chip with high sample volume loaded, as well. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Design, processing and testing of LSI arrays, hybrid microelectronics task

    NASA Technical Reports Server (NTRS)

    Himmel, R. P.; Stuhlbarg, S. M.; Ravetti, R. G.; Zulueta, P. J.; Rothrock, C. W.

    1979-01-01

    Mathematical cost models previously developed for hybrid microelectronic subsystems were refined and expanded. Rework terms related to substrate fabrication, nonrecurring developmental and manufacturing operations, and prototype production are included. Sample computer programs were written to demonstrate hybrid microelectric applications of these cost models. Computer programs were generated to calculate and analyze values for the total microelectronics costs. Large scale integrated (LST) chips utilizing tape chip carrier technology were studied. The feasibility of interconnecting arrays of LSU chips utilizing tape chip carrier and semiautomatic wire bonding technology was demonstrated.

  2. Integrated potentiometric detector for use in chip-based flow cells

    PubMed

    Tantra; Manz

    2000-07-01

    A new kind of potentiometric chip sensor for ion-selective electrodes (ISE) based on a solvent polymeric membrane is described. The chip sensor is designed to trap the organic cocktail inside the chip and to permit sample solution to flow past the membrane. The design allows the sensor to overcome technical problems of ruggedness and would therefore be ideal for industrial processes. The sensor performance for a Ba2+-ISE membrane based on a Vogtle ionophore showed electrochemical behavior similar to that observed in conventional electrodes and microelectrode arrangements.

  3. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Murray, E.; Floether, F. F.; Cavendish Laboratory, University of Cambridge, J.J. Thomson Avenue, Cambridge CB3 0HE

    Fundamental to integrated photonic quantum computing is an on-chip method for routing and modulating quantum light emission. We demonstrate a hybrid integration platform consisting of arbitrarily designed waveguide circuits and single-photon sources. InAs quantum dots (QD) embedded in GaAs are bonded to a SiON waveguide chip such that the QD emission is coupled to the waveguide mode. The waveguides are SiON core embedded in a SiO{sub 2} cladding. A tuneable Mach Zehnder interferometer (MZI) modulates the emission between two output ports and can act as a path-encoded qubit preparation device. The single-photon nature of the emission was verified using themore » on-chip MZI as a beamsplitter in a Hanbury Brown and Twiss measurement.« less

  4. Monitoring moisture content, temperature, and humidity in whole-tree pine chip piles

    Treesearch

    John Klepac; Dana Mitchell; Jason Thompson

    2015-01-01

    Two whole-tree chip piles were monitored for moisture content, temperature, and relative humidity from October 8th, 2010 to March 16th, 2011 at a location in south Alabama. Initial moisture content samples were collected immediately after chips were delivered to the study location on October 8th for Pile 1 and October 22nd for Pile 2. During pile construction, Lascar...

  5. Nanoliter-Scale Oil-Air-Droplet Chip-Based Single Cell Proteomic Analysis.

    PubMed

    Li, Zi-Yi; Huang, Min; Wang, Xiu-Kun; Zhu, Ying; Li, Jin-Song; Wong, Catherine C L; Fang, Qun

    2018-04-17

    Single cell proteomic analysis provides crucial information on cellular heterogeneity in biological systems. Herein, we describe a nanoliter-scale oil-air-droplet (OAD) chip for achieving multistep complex sample pretreatment and injection for single cell proteomic analysis in the shotgun mode. By using miniaturized stationary droplet microreaction and manipulation techniques, our system allows all sample pretreatment and injection procedures to be performed in a nanoliter-scale droplet with minimum sample loss and a high sample injection efficiency (>99%), thus substantially increasing the analytical sensitivity for single cell samples. We applied the present system in the proteomic analysis of 100 ± 10, 50 ± 5, 10, and 1 HeLa cell(s), and protein IDs of 1360, 612, 192, and 51 were identified, respectively. The OAD chip-based system was further applied in single mouse oocyte analysis, with 355 protein IDs identified at the single oocyte level, which demonstrated its special advantages of high enrichment of sequence coverage, hydrophobic proteins, and enzymatic digestion efficiency over the traditional in-tube system.

  6. A one-step strategy for ultra-fast and low-cost mass production of plastic membrane microfluidic chips.

    PubMed

    Hu, Chong; Lin, Sheng; Li, Wanbo; Sun, Han; Chen, Yangfan; Chan, Chiu-Wing; Leung, Chung-Hang; Ma, Dik-Lung; Wu, Hongkai; Ren, Kangning

    2016-10-05

    An ultra-fast, extremely cost-effective, and environmentally friendly method was developed for fabricating flexible microfluidic chips with plastic membranes. With this method, we could fabricate plastic microfluidic chips rapidly (within 12 seconds per piece) at an extremely low cost (less than $0.02 per piece). We used a heated perfluoropolymer perfluoroalkoxy (often called Teflon PFA) solid stamp to press a pile of two pieces of plastic membranes, low density polyethylene (LDPE) and polyethylene terephthalate (PET) coated with an ethylene-vinyl acetate copolymer (EVA). During the short period of contact with the heated PFA stamp, the pressed area of the membranes permanently bonded, while the LDPE membrane spontaneously rose up at the area not pressed, forming microchannels automatically. These two regions were clearly distinguishable even at the micrometer scale so we were able to fabricate microchannels with widths down to 50 microns. This method combines the two steps in the conventional strategy for microchannel fabrication, generating microchannels and sealing channels, into a single step. The production is a green process without using any solvent or generating any waste. Also, the chips showed good resistance against the absorption of Rhodamine 6G, oligonucleotides, and green fluorescent protein (GFP). We demonstrated some typical microfluidic manipulations with the flexible plastic membrane chips, including droplet formation, on-chip capillary electrophoresis, and peristaltic pumping for quantitative injection of samples and reagents. In addition, we demonstrated convenient on-chip detection of lead ions in water samples by a peristaltic-pumping design, as an example of the application of the plastic membrane chips in a resource-limited environment. Due to the high speed and low cost of the fabrication process, this single-step method will facilitate the mass production of microfluidic chips and commercialization of microfluidic technologies.

  7. Clonal haemopoiesis and therapy-related myeloid malignancies in elderly patients: a proof-of-concept, case-control study.

    PubMed

    Gillis, Nancy K; Ball, Markus; Zhang, Qing; Ma, Zhenjun; Zhao, YuLong; Yoder, Sean J; Balasis, Maria E; Mesa, Tania E; Sallman, David A; Lancet, Jeffrey E; Komrokji, Rami S; List, Alan F; McLeod, Howard L; Alsina, Melissa; Baz, Rachid; Shain, Kenneth H; Rollison, Dana E; Padron, Eric

    2017-01-01

    Clonal haemopoiesis of indeterminate potential (CHIP) is an age-associated genetic event linked to increased risk of primary haematological malignancies and increased all-cause mortality, but the prevalence of CHIP in patients who develop therapy-related myeloid neoplasms is unknown. We did this study to investigate whether chemotherapy-treated patients with cancer who have CHIP are at increased risk of developing therapy-related myeloid neoplasms. We did a nested, case-control, proof-of-concept study to compare the prevalence of CHIP between patients with cancer who later developed therapy-related myeloid neoplasms (cases) and patients who did not develop these neoplasms (controls). We identified cases from our internal biorepository of 123 357 patients who consented to participate in the Total Cancer Care biobanking protocol at Moffitt Cancer Center (Tampa, FL, USA) between Jan 1, 2006, and June 1, 2016. We included all individuals who were diagnosed with a primary malignancy, were treated with chemotherapy, subsequently developed a therapy-related myeloid neoplasm, and were 70 years or older at either diagnosis. For inclusion in this study, individuals must have had a peripheral blood or mononuclear cell sample collected before the diagnosis of therapy-related myeloid neoplasm. Controls were individuals who were diagnosed with a primary malignancy at age 70 years or older and were treated with chemotherapy but did not develop therapy-related myeloid neoplasms. Controls were matched to cases in at least a 4:1 ratio on the basis of sex, primary tumour type, age at diagnosis, smoking status, chemotherapy drug class, and duration of follow-up. We used sequential targeted and whole-exome sequencing and described clonal evolution in cases for whom paired CHIP and therapy-related myeloid neoplasm samples were available. The primary endpoint of this study was the development of therapy-related myeloid neoplasm and the primary exposure was CHIP. We identified 13 cases and 56 case-matched controls. The prevalence of CHIP in all patients (23 [33%] of 69 patients) was higher than has previously been reported in elderly individuals without cancer (about 10%). Cases had a significantly higher prevalence of CHIP than did matched controls (eight [62%] of 13 cases vs 15 [27%] of 56 controls, p=0·024; odds ratio 5·75, 95% CI 1·52-25·09, p=0·013). The most commonly mutated genes in cases with CHIP were TET2 (three [38%] of eight patients) and TP53(three [38%] of eight patients), whereas controls most often had TET2 mutations (six [40%] of 15 patients). In most (four [67%] of six patients) cases for whom paired CHIP and therapy-related myeloid neoplasm samples were available, the mean allele frequency of CHIP mutations had expanded by the time of the therapy-related myeloid neoplasm diagnosis. However, a subset of paired samples (two [33%] of six patients) had CHIP mutations that decreased in allele frequency, giving way to expansion of a distinct mutant clone. Patients with cancer who have CHIP are at increased risk of developing therapy-related myeloid neoplasms. The distribution of CHIP-related gene mutations differs between individuals with therapy-related myeloid neoplasm and those without, suggesting that mutation-specific differences might exist in therapy-related myeloid neoplasm risk. Moffitt Cancer Center. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Impact of pre-drying and frying time on physical properties and sensorial acceptability of fried potato chips.

    PubMed

    Cruz, Giorman; Cruz-Tirado, J P; Delgado, Kevin; Guzman, Yorvin; Castro, Franco; Rojas, Meliza Lindsay; Linares, Guillermo

    2018-01-01

    In this work the effects of pre-drying and frying time on colour, oil, texture and sensorial acceptability (overall liking) of potato chips were evaluated. Potato chips were pre-dried for 0, 10, 20 and 30 min at 60 °C and fried in soybean oil at 190 °C for 60, 70 and 80 s. The colour parameters (L*, a* and b*) increased or decreased depending on the pre-drying and frying time. Hardness increased as the pre-drying and frying time increased. On the other hand, the water initially removed by pre-drying decrease the gradient of mass transfer (water-oil). The oil content reduced to (about 21%) in pre-dried samples when compared to control sample. Finally, sensorial evaluation showed that samples without pre-drying and/or fried for very short or very long times had low acceptance levels. The pre-drying and frying times influenced the colour, texture, water and oil content, and resulted into fried potato chips with better acceptance scores.

  9. Infinium HumanMethylation450 BeadChip

    Cancer.gov

    The HumanMethylation450 BeadChip offers a unique combination of comprehensive, expert-selected coverage and high throughput at a low price, making it ideal for screening large sample populations such as those used in genome-wide association study cohorts. By providing quantitative methylation measurement at the single-CpG–site level for normal and FFPE samples, this assay offers powerful resolution for understanding epigenetic changes.

  10. Rapid antibiotic efficacy screening with aluminum oxide nanoporous membrane filter-chip and optical detection system.

    PubMed

    Tsou, Pei-Hsiang; Sreenivasappa, Harini; Hong, Sungmin; Yasuike, Masayuki; Miyamoto, Hiroshi; Nakano, Keiyo; Misawa, Takeyuki; Kameoka, Jun

    2010-09-15

    We have developed a filter-chip and optical detection system for rapid antibiotic efficacy screening. The filter-chip consisted of a 1-mL reservoir and an anodic aluminum oxide (AAO) nanoporous membrane. Sample solution with liquid growth media, bacteria, and antibiotics was incubated in the reservoir for a specific period of time. The number of live bacteria on the surface of membrane was counted after the incubation with antibiotics and filtration. Using this biosensing system, we have demonstrated a 1-h antibiotic screening for patients' clinical samples, significantly faster than the conventional antibiotic susceptibility tests that typically take more than 24h. This rapid screening nature makes the filter-chip and detection system ideal for tailoring antibiotic treatment to individual patients by reducing the microbial antibiotic resistance, and improving the survival rate for patients suffering from postoperative infections. Published by Elsevier B.V.

  11. Microfluidic Devices for Forensic DNA Analysis: A Review.

    PubMed

    Bruijns, Brigitte; van Asten, Arian; Tiggelaar, Roald; Gardeniers, Han

    2016-08-05

    Microfluidic devices may offer various advantages for forensic DNA analysis, such as reduced risk of contamination, shorter analysis time and direct application at the crime scene. Microfluidic chip technology has already proven to be functional and effective within medical applications, such as for point-of-care use. In the forensic field, one may expect microfluidic technology to become particularly relevant for the analysis of biological traces containing human DNA. This would require a number of consecutive steps, including sample work up, DNA amplification and detection, as well as secure storage of the sample. This article provides an extensive overview of microfluidic devices for cell lysis, DNA extraction and purification, DNA amplification and detection and analysis techniques for DNA. Topics to be discussed are polymerase chain reaction (PCR) on-chip, digital PCR (dPCR), isothermal amplification on-chip, chip materials, integrated devices and commercially available techniques. A critical overview of the opportunities and challenges of the use of chips is discussed, and developments made in forensic DNA analysis over the past 10-20 years with microfluidic systems are described. Areas in which further research is needed are indicated in a future outlook.

  12. A sensitive molecularly imprinted polymer based quartz crystal microbalance nanosensor for selective determination of lovastatin in red yeast rice.

    PubMed

    Eren, Tanju; Atar, Necip; Yola, Mehmet Lütfi; Karimi-Maleh, Hassan

    2015-10-15

    Lovastatin (LOV) is a statin, used to lower cholesterol which has been found as a hypolipidemic agent in commercial red yeast rice. In present study, a sensitive molecular imprinted quartz crystal microbalance (QCM) sensor was prepared by fabricating a self-assembling monolayer formation of allylmercaptane on QCM chip surface for selective determination of lovastatin (LOV) in red yeast rice. To prepare molecular imprinted quartz crystal microbalance (QCM) nanosensor, LOV imprinted poly(2-hydroxyethyl methacrylate-methacryloylamidoaspartic acid) [p(HEMA-MAAsp)] nanofilm was attached on the modified gold surface of QCM chip. The non-modified and improved surfaces were characterized by using contact angle, atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectroscopy. The imprinted QCM sensor was validated according to the ICH guideline (International Conference on Harmonisation). The linearity range was obtained as 0.10-1.25 nM. The detection limit of the prepared material was calculated as 0.030 nM. The developed QCM nanosensor was successfully used to examine red yeast rice. Furthermore, the stability and repeatability of the prepared QCM nanosensor were studied. The spectacular long-term stability and repeatability of the prepared LOV-imprinted QCM nanosensor make them intriguing for use in QCM sensors. Copyright © 2015 Elsevier Ltd. All rights reserved.

  13. Smartphone-enabled optofluidic exosome diagnostic for concussion recovery

    NASA Astrophysics Data System (ADS)

    Ko, Jina; Hemphill, Matthew A.; Gabrieli, David; Wu, Leon; Yelleswarapu, Venkata; Lawrence, Gladys; Pennycooke, Wesley; Singh, Anup; Meaney, Dave F.; Issadore, David

    2016-08-01

    A major impediment to improving the treatment of concussion is our current inability to identify patients that will experience persistent problems after the injury. Recently, brain-derived exosomes, which cross the blood-brain barrier and circulate following injury, have shown great potential as a noninvasive biomarker of brain recovery. However, clinical use of exosomes has been constrained by their small size (30-100 nm) and the extensive sample preparation (>24 hr) needed for traditional exosome measurements. To address these challenges, we developed a smartphone-enabled optofluidic platform to measure brain-derived exosomes. Sample-to-answer on our chip is 1 hour, 10x faster than conventional techniques. The key innovation is an optofluidic device that can detect enzyme amplified exosome biomarkers, and is read out using a smartphone camera. Using this approach, we detected and profiled GluR2+ exosomes in the post-injury state using both in vitro and murine models of concussion.

  14. Smartphone-enabled optofluidic exosome diagnostic for concussion recovery.

    PubMed

    Ko, Jina; Hemphill, Matthew A; Gabrieli, David; Wu, Leon; Yelleswarapu, Venkata; Lawrence, Gladys; Pennycooke, Wesley; Singh, Anup; Meaney, Dave F; Issadore, David

    2016-08-08

    A major impediment to improving the treatment of concussion is our current inability to identify patients that will experience persistent problems after the injury. Recently, brain-derived exosomes, which cross the blood-brain barrier and circulate following injury, have shown great potential as a noninvasive biomarker of brain recovery. However, clinical use of exosomes has been constrained by their small size (30-100 nm) and the extensive sample preparation (>24 hr) needed for traditional exosome measurements. To address these challenges, we developed a smartphone-enabled optofluidic platform to measure brain-derived exosomes. Sample-to-answer on our chip is 1 hour, 10x faster than conventional techniques. The key innovation is an optofluidic device that can detect enzyme amplified exosome biomarkers, and is read out using a smartphone camera. Using this approach, we detected and profiled GluR2+ exosomes in the post-injury state using both in vitro and murine models of concussion.

  15. Automated payload and instruments for astrobiology research developed and studied by German medium-sized space industry in cooperation with European academia

    NASA Astrophysics Data System (ADS)

    Schulte, Wolfgang; Hofer, Stefan; Hofmann, Peter; Thiele, Hans; von Heise-Rotenburg, Ralf; Toporski, Jan; Rettberg, Petra

    2007-06-01

    For more than a decade Kayser-Threde, a medium-sized enterprise of the German space industry, has been involved in astrobiology research in partnership with a variety of scientific institutes from all over Europe. Previous projects include exobiology research platforms in low Earth orbit on retrievable carriers and onboard the Space Station. More recently, exobiology payloads for in situ experimentation on Mars have been studied by Kayser-Threde under ESA contracts, specifically the ExoMars Pasteur Payload. These studies included work on a sample preparation and distribution systems for Martian rock/regolith samples, instrument concepts such as Raman spectroscopy and a Life Marker Chip, advanced microscope systems as well as robotic tools for astrobiology missions. The status of the funded technical studies and major results are presented. The reported industrial work was funded by ESA and the German Aerospace Center (DLR).

  16. Fabrication of an inexpensive, implantable cooling device for reversible brain deactivation in animals ranging from rodents to primates

    PubMed Central

    Cooke, Dylan F.; Goldring, Adam B.; Yamayoshi, Itsukyo; Tsourkas, Phillippos; Recanzone, Gregg H.; Tiriac, Alex; Pan, Tingrui; Simon, Scott I.

    2012-01-01

    We have developed a compact and lightweight microfluidic cooling device to reversibly deactivate one or more areas of the neocortex to examine its functional macrocircuitry as well as behavioral and cortical plasticity. The device, which we term the “cooling chip,” consists of thin silicone tubing (through which chilled ethanol is circulated) embedded in mechanically compliant polydimethylsiloxane (PDMS). PDMS is tailored to compact device dimensions (as small as 21 mm3) that precisely accommodate the geometry of the targeted cortical area. The biocompatible design makes it suitable for both acute preparations and chronic implantation for long-term behavioral studies. The cooling chip accommodates an in-cortex microthermocouple measuring local cortical temperature. A microelectrode may be used to record simultaneous neural responses at the same location. Cortex temperature is controlled by computer regulation of the coolant flow, which can achieve a localized cortical temperature drop from 37 to 20°C in less than 3 min and maintain target temperature to within ±0.3°C indefinitely. Here we describe cooling chip fabrication and performance in mediating cessation of neural signaling in acute preparations of rodents, ferrets, and primates. PMID:22402651

  17. Handheld analyzer with on-chip molecularly-imprinted biosensors for electrical detection of propofol in plasma samples.

    PubMed

    Hong, Chien-Chong; Lin, Chih-Chung; Hong, Chian-Lang; Lin, Zi-Xiang; Chung, Meng-Hua; Hsieh, Pei-Wen

    2016-12-15

    This paper proposes a novel handheld analyzer with disposable lab-on-a-chip technology for the electrical detection of the anesthetic propofol in human plasma samples for clinical diagnoses. The developed on-chip biosensors are based on the conduction of molecularly imprinted polymers (MIPs) that employ label-free electrical detection techniques. Propofol in total intravenous anesthesia is widely used with a target-controlled infusion system. At present, the methods employed for detecting blood propofol concentrations in hospitals comprise high-performance liquid chromatography and ion mobility spectrometry. These conventional instruments are bulky, expensive, and difficult to access. In this study, we developed a novel plastic microfluidic biochip with an on-chip anesthetic biosensor that was characterized for the rapid detection of propofol concentrations. The experimental results revealed that the response time of the developed propofol biosensors was 25s. The specific binding of an MIP to a nonimprinted polymer (NIP) reached up to 560%. Moreover, the detection limit of the biosensors was 0.1μg/mL, with a linear detection range of 0.1-30μg/mL. The proposed disposable microfluidic biochip with an on-chip anesthetic biosensor using MIPs exhibited excellent performance in the separation and sensing of propofol molecules in the human plasma samples. Compared with large-scale conventional instruments, the developed microfluidic biochips with on-chip MIP biosensors present the advantages of a compact size, high selectivity, low cost, rapid response, and single-step detection. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Dry matter losses and quality changes during short rotation coppice willow storage in chip or rod form.

    PubMed

    Whittaker, Carly; Yates, Nicola E; Powers, Stephen J; Misselbrook, Tom; Shield, Ian

    2018-05-01

    This study compares dry matter losses and quality changes during the storage of SRC willow as chips and as rods. A wood chip stack consisting of approximately 74 tonnes of fresh biomass, or 31 tonnes dry matter (DM) was built after harvesting in the spring. Three weeks later, four smaller stacks of rods with an average weight of 0.8 tonnes, or 0.4 tonnes DM were built. During the course of the experiment temperature recorders placed in the stacks found that the wood chip pile reached 60 °C within 10 days of construction, but the piles of rods remained mostly at ambient temperatures. Dry matter losses were calculated by using pre-weighed independent samples within the stacks and by weighing the whole stack before and after storage. After 6 months the wood chip stack showed a DM loss of between 19.8 and 22.6%, and mean losses of 23.1% were measured from the 17 independent samples. In comparison, the rod stacks showed an average stack DM loss of between 0 and 9%, and between 1.4% and 10.6% loss from the independent samples. Analysis of the stored material suggests that storing willow in small piles of rods produces a higher quality fuel in terms of lower moisture and ash content; however, it has a higher fine content compared to storage in chip form. Therefore, according to the two storage methods tested here, there may be a compromise between maximising the net dry matter yield from SRC willow and the final fine content of the fuel.

  19. [Fabrications of a poly (methyl methacrylate) (PMMA) microfluidic chip-based DNA analysis device].

    PubMed

    Du, Xiao-Guang

    2009-12-01

    A DNA analysis device based on poly(methyl methacrylate) (PMMA) microfluidic chips was developed. A PMMA chip with cross microchannels was fabricated by a simple hot embossing. Microchannels were modified with a static adsorptive coating method using 2% hydroxyethyl cellulose. A high-voltage power unit, variable in the range 0-1 800 V, was used for on-chip DNA sample injection and gel electrophoretic separation. High speed, high resolution DNA analysis was obtained with the home-built PMMA chip in a sieving matrix containing 2% hydroxyethyl cellulose with a blue intercalating dye, TO-PRO-3 (TP3), by using diode laser induced fluorescence detection based on optical fibers with a 670 nm long-pass filter. The DNA analysis device was applied for the separation of phiX-174/HaeIII DNA digest sample with 11 fragments ranging from 72 to 1 353 bp. A separation efficiency of 1.14 x 10(6) plates/m was obtained for the 603 bp fragments, while the R of 271/281 bp fragments was 1.2. The device was characterized by simple design, low cost for fabrication and operation, reusable PMMA chips, and good reproducibility. A portable microfluidic device for DNA analysis can be developed for clinical diagnosis and disease screening.

  20. Sputtered Pd as Hydrogen Storage for a Chip-Integrated Microenergy System

    PubMed Central

    Slavcheva, E.; Ganske, G.; Schnakenberg, U.

    2014-01-01

    The work presents a research on preparation and physical and electrochemical characterisation of dc magnetron sputtered Pd films envisaged for application as hydrogen storage in a chip-integrated hydrogen microenergy system. The influence of the changes in the sputtering pressure on the surface structure, morphology, and roughness was analysed by X-ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AMF). The electrochemical activity towards hydrogen adsorption/desorption and formation of PdH were investigated in 0.5 M H2SO4 using the methods of cyclic voltammetry and galvanostatic polarisation. The changes in the electrical properties of the films as a function of the sputtering pressure and the level of hydrogenation were evaluated before and immediately after the electrochemical charging tests, using a four-probe technique. The research resulted in establishment of optimal sputter regime, ensuring fully reproducible Pd layers with highly developed surface, moderate porosity, and mechanical stability. Selected samples were integrated as hydrogen storage in a newly developed unitized microenergy system and tested in charging (water electrolysis) and discharging (fuel cell) operative mode at ambient conditions demonstrating a stable recycling performance. PMID:24516356

  1. Mode and polarization state selected guided wave spectroscopy of orientational anisotrophy in model membrane cellulosic polymer films: relevance to lab-on-a-chip

    NASA Astrophysics Data System (ADS)

    Andrews, Mark P.; Kanigan, Tanya

    2007-06-01

    Orientation anisotropies in structural properties relevant to the use of cellulosic polymers as membranes for lab-on-chips were investigated for cellulose acetate (CA) and regenerated cellulose (RC) films deposited as slab waveguides. Anisotropy was probed with mode and polarization state selected guided wave Raman spectroscopy. CA exhibits partial chain orientation in the plane of the film, and this orientation is independent of sample substrate and film preparation conditions. RC films also show in-plane anisotropy, where the hexose sugar rings lie roughly in the plane of the film. Explanations are given of the role of artifacts in interpreting waveguide Raman spectra, including anomalous contributions to Raman spectra that arise from deviations from right angle scattering geometry, mode-dependent contributions to longitudinal electric field components and TE<-->TM mode conversion. We explore diffusion profiles of small molecules in cellulosic films by adaptations of an inverse-Wentzel-Kramers-Brillouin (iWKB) recursive, noninteger virtual mode index algorithm. Perturbations in the refractive index distribution, n(z), are recovered from the measured relative propagation constants, neffective,m, of the planar waveguide. The refractive index distribution then yields the diffusion profile.

  2. Lack of mutagens in deep-fat-fried foods obtained at the retail level.

    PubMed

    Taylor, S L; Berg, C M; Shoptaugh, N H; Scott, V N

    1982-04-01

    The basic methylene chloride extract from 20 of 30 samples of foods fried in deep fat failed to elicit any mutagenic response that could be detected in the Salmonella typhimurium/mammalian microsome assay. The basic extracts of the remaining ten samples (all three chicken samples studied, two of the four potato-chip samples, one of four corn-chip samples, the sample of onion rings, two of six doughnuts, and one of three samples of french-fried potato) showed evidence of weak mutagenic activity. In these samples, amounts of the basic extract equivalent to 28.5-57 g of the original food sample were required to produce revertants at levels of 2.6-4.8 times the background level. Only two of the acidic methylene chloride extracts from the 30 samples exhibited mutagenic activity greater than 2.5 times the background reversion level, and in both cases (one corn-chip and one shrimp sample) the mutagenic response was quite weak. The basic extract of hamburgers fried in deep fat in a home-style fryer possessed higher levels of mutagenic activity (13 times the background reversion level). However, the mutagenic activity of deep-fried hamburgers is some four times lower than that of pan-fried hamburgers.

  3. RUNNER BOX MAINTENANCE. FRANK FEHER USES A COMPRESSED AIRPOWERED CHISEL ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    RUNNER BOX MAINTENANCE. FRANK FEHER USES A COMPRESSED AIR-POWERED CHISEL TO CHIP OUT CONGEALED METAL IN PREPARATION FOR ANOTHER HEAT. - American Brass Foundry, 70 Sayre Street, Buffalo, Erie County, NY

  4. On-chip wavelength multiplexed detection of cancer DNA biomarkers in blood

    PubMed Central

    Cai, H.; Stott, M. A.; Ozcelik, D.; Parks, J. W.; Hawkins, A. R.; Schmidt, H.

    2016-01-01

    We have developed an optofluidic analysis system that processes biomolecular samples starting from whole blood and then analyzes and identifies multiple targets on a silicon-based molecular detection platform. We demonstrate blood filtration, sample extraction, target enrichment, and fluorescent labeling using programmable microfluidic circuits. We detect and identify multiple targets using a spectral multiplexing technique based on wavelength-dependent multi-spot excitation on an antiresonant reflecting optical waveguide chip. Specifically, we extract two types of melanoma biomarkers, mutated cell-free nucleic acids —BRAFV600E and NRAS, from whole blood. We detect and identify these two targets simultaneously using the spectral multiplexing approach with up to a 96% success rate. These results point the way toward a full front-to-back chip-based optofluidic compact system for high-performance analysis of complex biological samples. PMID:28058082

  5. Determination of ammonium in wastewaters by capillary electrophoresis on a column-coupling chip with conductivity detection.

    PubMed

    Luc, Milan; Kruk, Pavol; Masár, Marián

    2011-07-01

    Analytical potentialities of a chip-based CE in determination of ammonium in wastewaters were investigated. CZE with the electric field and/or ITP sample stacking was performed on a column-coupling (CC) chip with integrated conductivity detectors. Acetate background electrolytes (pH ∼3) including 18-crown-6-ether (18-crown-6) and tartaric acid were developed to reach rapid (in 7-8 min) CZE and ITP-CZE resolutions of ammonium from other cations (sodium, potassium, calcium and magnesium) present in wastewater samples. Under preferred working conditions (suppressed hydrodynamic flow (HDF) and EOF on the column-coupling chip), both the employed methods did provide very good repeatabilities of the migration (RSD of 0.2-0.8% for the migration time) and quantitative (RSD of 0.3-4.9% for the peak area) parameters in the model and wastewater samples. Using a 900-nL sample injection volume, LOD for ammonium were obtained at 20 and 40 μg/L concentrations in CZE and ITP-CZE separations, respectively. Very good agreements of the CZE and ITP-CZE determinations of ammonium in six untreated wastewater samples (only filtration and dilution) with the results obtained by a reference spectrometric method indicate a very good accuracy of both the CE methods presented. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Integrated chemical/biochemical sample collection, pre-concentration, and analysis on a digital microfluidic lab-on-a-chip platform

    NASA Astrophysics Data System (ADS)

    Fair, Richard B.; Khlystov, A.; Srinivasan, Vijay; Pamula, Vamsee K.; Weaver, Kathryn N.

    2004-12-01

    An ideal on-site chemical/biochemical analysis system must be inexpensive, sensitive, fully automated and integrated, reliable, and compatible with a broad range of samples. The advent of digital microfluidic lab-on-a-chip (LoC) technology offers such a detection system due to the advantages in portability, reduction of the volumes of the sample and reagents, faster analysis times, increased automation, low power consumption, compatibility with mass manufacturing, and high throughput. We describe progress towards integrating sample collection onto a digital microfluidic LoC that is a component of a cascade impactor device. The sample collection is performed by impacting airborne particles directly onto the surface of the chip. After the collection phase, the surface of the chip is washed with a micro-droplet of solvent. The droplet will be digitally directed across the impaction surface, dissolving sample constituents. Because of the very small droplet volume used for extraction of the sample from a wide colection area, the resulting solution is realatively concentrated and the analytes can be detected after a very short sampling time (1 min) due to such pre-concentration. After the washing phase, the droplet is mixed with specific reagents that produce colored reaction products. The concentration of the analyte is quantitatively determined by measuring absorption at target wavelengths using a simple light emitting diode and photodiode setup. Specific applications include automatic measurements of major inorganic ions in aerosols, such as sulfate, nitrate and ammonium, with a time resolution of 1 min and a detection limit of 30 nm/m3. We have already demonstrated the detection and quantification of nitroaromatic explosives without integrating the sample collection. Other applications being developed include airborne bioagent detection.

  7. Digital PCR on an integrated self-priming compartmentalization chip.

    PubMed

    Zhu, Qiangyuan; Qiu, Lin; Yu, Bingwen; Xu, Yanan; Gao, Yibo; Pan, Tingting; Tian, Qingchang; Song, Qi; Jin, Wei; Jin, Qinhan; Mu, Ying

    2014-03-21

    An integrated on-chip valve-free and power-free microfluidic digital PCR device is for the first time developed by making use of a novel self-priming compartmentalization and simple dehydration control to realize 'divide and conquer' for single DNA molecule detection. The high gas solubility of PDMS is exploited to provide the built-in power of self-priming so that the sample and oil are sequentially sucked into the device to realize sample self-compartmentalization based on surface tension. The lifespan of its self-priming capability was about two weeks tested using an air-tight packaging bottle sealed with a small amount of petroleum jelly, which is significant for a practical platform. The SPC chip contains 5120 independent 5 nL microchambers, allowing the samples to be compartmentalized completely. Using this platform, three different abundances of lung cancer related genes are detected to demonstrate the feasibility and flexibility of the microchip for amplifying a single nucleic acid molecule. For maximal accuracy, within less than 5% of the measurement deviation, the optimal number of positive chambers is between 400 and 1250 evaluated by the Poisson distribution, which means one panel can detect an average of 480 to 4804 template molecules. This device without world-to-chip connections eliminates the constraint of the complex pipeline control, and is an integrated on-chip platform, which would be a significant improvement to digital PCR automation and more user-friendly.

  8. Gastrointestinal Illness Associated with Rancid Tortilla Chips at a Correctional Facility - Wyoming, 2015.

    PubMed

    Lupcho, Tiffany; Harrist, Alexia; Van Houten, Clay

    2016-10-28

    On October 12, 2015, a county health department notified the Wyoming Department of Health of an outbreak of gastrointestinal illness among residents and staff members at a local correctional facility. The majority of ill persons reported onset of symptoms within 1-3 hours after eating lunch served at the facility cafeteria at noon on October 11. Residents and staff members reported that tortilla chips served at the lunch tasted and smelled like chemicals. The Wyoming Department of Health and county health department personnel conducted case-control studies to identify the outbreak source. Consuming lunch at the facility on October 11 was highly associated with illness; multivariate logistic regression analysis found that tortilla chips were the only food item associated with illness. Hexanal and peroxide, markers for rancidity, were detected in tortilla chips and composite food samples from the lunch. No infectious agent was detected in human stool specimens or food samples. Extensive testing of lunch items did not identify any unusual chemical. Epidemiologic and laboratory evidence implicated rancid tortilla chips as the most likely source of illness. This outbreak serves as a reminder to consider alternative food testing methods during outbreaks of unusual gastrointestinal illness when typical foodborne pathogens are not identified. For interpretation of alternative food testing results, samples of each type of food not suspected to be contaminated are needed to serve as controls.

  9. Acrylamide formation in plantain (Musa paradisiaca) chips influenced by different ripening stages: A correlation study with respect to reducing sugars, amino acids and phenolic content.

    PubMed

    Shamla, L; Nisha, P

    2017-05-01

    The effect of ripening on the formation of acrylamide in deep fried plantain chips made from Nendran variety (Musa paradisiaca) was investigated. The precursors of acrylamide formation, reducing sugars (glucose and fructose) and ten major amino acids, were quantified during different stages of ripening using HPLC and correlated with acrylamide formation. The total phenolic content and total flavonoid content were also estimated and correlated with acrylamide formation. Both glucose and fructose increased during ripening and demonstrated a positive correlation on formation of acrylamide (correlation coefficient of r=0.95 and 0.94 respectively (p<0.05), whereas asparagine, was poorly correlated (p>0.05). The decreased levels of phenolic content during ripening of plantain were negatively correlated with acrylamide formation in the deep fried chips prepared. Thus the selection of proper ripening stage renders reduced formation of acrylamide in plantain chips to a reasonable extend. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. A sensitive electrochemical immunosensor for label-free detection of Zika-virus protein.

    PubMed

    Kaushik, Ajeet; Yndart, Adriana; Kumar, Sanjeev; Jayant, Rahul Dev; Vashist, Arti; Brown, Ashley N; Li, Chen-Zhong; Nair, Madhavan

    2018-06-26

    This work, as a proof of principle, presents a sensitive and selective electrochemical immunosensor for Zika-virus (ZIKV)-protein detection using a functionalized interdigitated micro-electrode of gold (IDE-Au) array. A miniaturized IDE-Au immunosensing chip was prepared via immobilization of ZIKV specific envelop protein antibody (Zev-Abs) onto dithiobis(succinimidyl propionate) i.e., (DTSP) functionalized IDE-Au (electrode gap/width of 10 µm). Electrochemical impedance spectroscopy (EIS) was performed to measure the electrical response of developed sensing chip as a function of ZIKV-protein concentrations. The results of EIS studies confirmed that sensing chip detected ZIKV-protein selectively and exhibited a detection range from 10 pM to 1 nM and a detection limit of 10 pM along with a high sensitivity of 12 kΩM -1 . Such developed ZIKV immune-sensing chip can be integrated with a miniaturized potentiostat (MP)-interfaced with a smartphone for rapid ZIKV-infection detection required for early stage diagnostics at point-of-care application.

  11. Considerations on Experimental Design and Data Analysis of Chromatin Immunoprecipitation Experiments.

    PubMed

    Jordán-Pla, Antonio; Visa, Neus

    2018-01-01

    Arguably one of the most valuable techniques to study chromatin organization, ChIP is the method of choice to map the contacts established between proteins and genomic DNA. Ever since its inception, more than 30 years ago, ChIP has been constantly evolving, improving, and expanding its capabilities and reach. Despite its widespread use by many laboratories across a wide variety of disciplines, ChIP assays can be sometimes challenging to design, and are often sensitive to variations in practical implementation.In this chapter, we provide a general overview of the ChIP method and its most common variations, with a special focus on ChIP-seq. We try to address some of the most important aspects that need to be taken into account in order to design and perform experiments that generate the most reproducible, high-quality data. Some of the main topics covered include the use of properly characterized antibodies, alternatives to chromatin preparation, the need for proper controls, and some recommendations about ChIP-seq data analysis.

  12. Application of infrared reflection and Raman spectroscopy for quantitative determination of fat in potato chips

    NASA Astrophysics Data System (ADS)

    Mazurek, Sylwester; Szostak, Roman; Kita, Agnieszka

    2016-12-01

    Potato chips are important products in the snack industry. The most significant parameter monitored during their quality control process is fat content. The Soxhlet method, which is applied for this purpose, is time consuming and expensive. We demonstrate that both infrared and Raman spectroscopy can effectively replace the extraction method. Raman, mid-infrared (MIR) and near-infrared (NIR) spectra of the homogenised laboratory-prepared chips were recorded. On the basis of obtained spectra, partial least squares (PLS) calibration models were constructed. They were characterised by the values of relative standard errors of prediction (RSEP) in the 1.0-1.9% range for both calibration and validation data sets. Using the developed models, six commercial products were successfully quantified with recovery in the 98.5-102.3% range against the AOAC extraction method. The proposed method for fat quantification in potato chips based on Raman spectroscopy can be easily adopted for on-line product analysis.

  13. Quantitative aspects of microchip isotachophoresis for high precision determination of main components in pharmaceuticals.

    PubMed

    Hradski, Jasna; Chorváthová, Mária Drusková; Bodor, Róbert; Sabo, Martin; Matejčík, Štefan; Masár, Marián

    2016-12-01

    Although microchip electrophoresis (MCE) is intended to provide reliable quantitative data, so far there is only limited attention paid to these important aspects. This study gives a general overview of key aspects to be followed to reach high-precise determination using isotachophoresis (ITP) on the microchip with conductivity detection. From the application point of view, the procedure for the determination of acetate, a main component in the pharmaceutical preparation buserelin acetate, was developed. Our results document that run-to-run fluctuations in the sample injection volume limit the reproducibility of quantitation based on the external calibration. The use of a suitable internal standard (succinate in this study) improved the repeatability of the precision of acetate determination from six to eight times. The robustness of the procedure was studied in terms of impact of fluctuations in various experimental parameters (driving current, concentration of the leading ions, pH of the leading electrolyte and buffer impurities) on the precision of the ITP determination. The use of computer simulation programs provided means to assess the ITP experiments using well-defined theoretical models. A long-term validity of the calibration curves on two microchips and two MCE equipments was verified. This favors ITP over other microchip electrophoresis techniques, when chip-to-chip or equipment-to-equipment transfer of the analytical method is required. The recovery values in the range of 98-101 % indicate very accurate determination of acetate in buserelin acetate, which is used in the treatment of hormone-dependent tumors. This study showed that microchip ITP is suitable for reliable determination of main components in pharmaceutical preparations.

  14. miRNA detection at single-cell resolution using microfluidic LNA flow-FISH

    DOE PAGES

    Wu, Meiye; Piccini, Matthew Ernest; Koh, Chung -Yan; ...

    2014-08-20

    Flow cytometry in combination with fluorescent in situ hybridization (flow-FISH) is a powerful technique that can be utilized to rapidly detect nucleic acids at single-cell resolution without the need for homogenization or nucleic acid extraction. Here, we describe a microfluidic-based method which enables the detection of microRNAs or miRNAs in single intact cells by flow-FISH using locked nucleic acid (LNA)-containing probes. Our method can be applied to all RNA species including mRNA and small noncoding RNA and is suitable for multiplexing with protein immunostaining in the same cell. For demonstration of our method, this chapter details the detection of miR155more » and CD69 protein in PMA and ionomycin-stimulated Jurkat cells. Here, we also include instructions on how to set up a microfluidic chip sample preparation station to prepare cells for imaging and analysis on a commercial flow cytometer or a custom-built micro-flow cytometer.« less

  15. Embellishment of microfluidic devices via femtosecond laser micronanofabrication for chip functionalization.

    PubMed

    Wang, Juan; He, Yan; Xia, Hong; Niu, Li-Gang; Zhang, Ran; Chen, Qi-Dai; Zhang, Yong-Lai; Li, Yan-Feng; Zeng, Shao-Jiang; Qin, Jian-Hua; Lin, Bing-Cheng; Sun, Hong-Bo

    2010-08-07

    This paper demonstrates the embellishment of existing microfluidic devices with integrated three dimensional (3D) micronanostructures via femtosecond laser micronanofabrication, which, for the first time, proves two-photon photopolymerization (TPP) to be a powerful technology for chip functionalization. As representative examples, microsieves with various pore shape and adjustable pore size were successfully fabricated inside a conventional glass-based microfluidic channel prepared by wet etching for microparticle separation. Moreover, a fish scale like microfilter was also fabricated and appointed as a one-way valve, which showed excellent performance as we expected. These results indicate that such embellishment of microfluidic devices is simple, low cost, flexible and easy to access. We believe that, combined with TPP, the application of lab-on-chip devices would be further extended.

  16. Modeling and Simulation of Lab-on-a-Chip Systems

    DTIC Science & Technology

    2005-08-12

    complex chip geometries (including multiple turns). Variations of sample concentration profiles in laminar diffusion-based micromixers are also derived...CHAPTER 6 MODELING OF LAMINAR DIFFUSION-BASED COMPLEX ELECTROKINETIC PASSIVE MICROMIXERS ...140 6.4.4 Multi-Stream (Inter-Digital) Micromixers

  17. Biostability of an implantable glucose sensor chip

    NASA Astrophysics Data System (ADS)

    Fröhlich, M.; Birkholz, M.; Ehwald, K. E.; Kulse, P.; Fursenko, O.; Katzer, J.

    2012-12-01

    Surface materials of an implantable microelectronic chip intended for medical applications were evaluated with respect to their long-term stability in bio-environments. The sensor chip shall apply in a glucose monitor by operating as a microviscosimeter according to the principle of affinity viscosimetry. A monolithic integration of a microelectromechanical system (MEMS) into the sensor chip was successfully performed in a combined 0.25 μm CMOS/BiCMOS technology. In order to study material durability and biostability of the surfaces, sensor chips were exposed to various in vitro and in vivo tests. Corrosional damage of SiON, SiO2 and TiN surfaces was investigated by optical microscopy, ellipsometry and AFM. The results served for optimizing the Back-end-of-Line (BEoL) stack, from which the MEMS was prepared. Corrosion of metal lines could significantly be reduced by improving the topmost passivation layer. The experiments revealed no visible damage of the actuator or other functionally important MEMS elements. Sensor chips were also exposed to human body fluid for three month by implantation into the abdomen of a volunteer. Only small effects were observed for layer thickness and Ra roughness after explantation. In particular, TiN as used for the actuator beam showed no degradation by biocorrosion. The highest degradation rate of about 50 nm per month was revealed for the SiON passivation layer. These results suggest that the sensor chip may safely operate in subcutaneous tissue for a period of several months.

  18. Size, shape and flow characterization of ground wood chip and ground wood pellet particles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rezaei, Hamid; Lim, C. Jim; Lau, Anthony

    Size, shape and density of biomass particles influence their transportation, fluidization, rates of drying and thermal decomposition. Pelleting wood particles increases the particle density and reduces the variability of physical properties among biomass particles. In this study, pine chips prepared for pulping and commercially produced pine pellets were ground in a hammer mill using grinder screens of 3.2, 6.3, 12.7 and 25.4mmperforations. Pellets consumed about 7 times lower specific grinding energy than chips to produce the same size of particles. Grinding pellets produced the smaller particles with narrower size distribution than grinding chips. Derived shape factors in digital image analysismore » showed that chip particles were rectangular and had the aspect ratios about one third of pellet particles. Pellet particles were more circular shape. The mechanical sieving underestimated the actual particle size and did not represent the size of particles correctly. Instead, digital imaging is preferred. Angle of repose and compressibility tests represented the flow properties of ground particles. Pellet particles made a less compacted bulk, had lower cohesion and did flow easier in a pile of particles. In conclusion, particle shape affected the flow properties more than particle size« less

  19. Size, shape and flow characterization of ground wood chip and ground wood pellet particles

    DOE PAGES

    Rezaei, Hamid; Lim, C. Jim; Lau, Anthony; ...

    2016-07-11

    Size, shape and density of biomass particles influence their transportation, fluidization, rates of drying and thermal decomposition. Pelleting wood particles increases the particle density and reduces the variability of physical properties among biomass particles. In this study, pine chips prepared for pulping and commercially produced pine pellets were ground in a hammer mill using grinder screens of 3.2, 6.3, 12.7 and 25.4mmperforations. Pellets consumed about 7 times lower specific grinding energy than chips to produce the same size of particles. Grinding pellets produced the smaller particles with narrower size distribution than grinding chips. Derived shape factors in digital image analysismore » showed that chip particles were rectangular and had the aspect ratios about one third of pellet particles. Pellet particles were more circular shape. The mechanical sieving underestimated the actual particle size and did not represent the size of particles correctly. Instead, digital imaging is preferred. Angle of repose and compressibility tests represented the flow properties of ground particles. Pellet particles made a less compacted bulk, had lower cohesion and did flow easier in a pile of particles. In conclusion, particle shape affected the flow properties more than particle size« less

  20. Advanced platform for the in-plane ZT measurement of thin films

    NASA Astrophysics Data System (ADS)

    Linseis, V.; Völklein, F.; Reith, H.; Nielsch, K.; Woias, P.

    2018-01-01

    The characterization of nanostructured samples with at least one restricted dimension like thin films or nanowires is challenging, but important to understand their structure and transport mechanism, and to improve current industrial products and production processes. We report on the 2nd generation of a measurement chip, which allows for a simplified sample preparation process, and the measurement of samples deposited from the liquid phase using techniques like spin coating and drop casting. The new design enables us to apply much higher temperature gradients for the Seebeck coefficient measurement in a shorter time, without influencing the sample holder's temperature distribution. Furthermore, a two membrane correction method for the 3ω thermal conductivity measurement will be presented, which takes the heat loss due to radiation into account and increases the accuracy of the measurement results significantly. Errors caused by different sample compositions, varying sample geometries, and different heat profiles are avoided with the presented measurement method. As a showcase study displaying the validity and accuracy of our platform, we present temperature-dependent measurements of the thermoelectric properties of an 84 nm Bi87Sb13 thin film and a 15 μm PEDOT:PSS thin film.

  1. Estimation and partitioning of (co)heritability of inflammatory bowel disease from GWAS and immunochip data.

    PubMed

    Chen, Guo-Bo; Lee, Sang Hong; Brion, Marie-Jo A; Montgomery, Grant W; Wray, Naomi R; Radford-Smith, Graham L; Visscher, Peter M

    2014-09-01

    As custom arrays are cheaper than generic GWAS arrays, larger sample size is achievable for gene discovery. Custom arrays can tag more variants through denser genotyping of SNPs at associated loci, but at the cost of losing genome-wide coverage. Balancing this trade-off is important for maximizing experimental designs. We quantified both the gain in captured SNP-heritability at known candidate regions and the loss due to imperfect genome-wide coverage for inflammatory bowel disease using immunochip (iChip) and imputed GWAS data on 61,251 and 38.550 samples, respectively. For Crohn's disease (CD), the iChip and GWAS data explained 19 and 26% of variation in liability, respectively, and SNPs in the densely genotyped iChip regions explained 13% of the SNP-heritability for both the iChip and GWAS data. For ulcerative colitis (UC), the iChip and GWAS data explained 15 and 19% of variation in liability, respectively, and the dense iChip regions explained 10 and 9% of the SNP-heritability in the iChip and the GWAS data. From bivariate analyses, estimates of the genetic correlation in risk between CD and UC were 0.75 (SE 0.017) and 0.62 (SE 0.042) for the iChip and GWAS data, respectively. We also quantified the SNP-heritability of genomic regions that did or did not contain the previous 163 GWAS hits for CD and UC, and SNP-heritability of the overlapping loci between the densely genotyped iChip regions and the 163 GWAS hits. For both diseases, over different genomic partitioning, the densely genotyped regions on the iChip tagged at least as much variation in liability as in the corresponding regions in the GWAS data, however a certain amount of tagged SNP-heritability in the GWAS data was lost using the iChip due to the low coverage at unselected regions. These results imply that custom arrays with a GWAS backbone will facilitate more gene discovery, both at associated and novel loci. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  2. Method of mechanical holding of cantilever chip for tip-scan high-speed atomic force microscope

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fukuda, Shingo; Uchihashi, Takayuki; Ando, Toshio

    In tip-scan atomic force microscopy (AFM) that scans a cantilever chip in the three dimensions, the chip body is held on the Z-scanner with a holder. However, this holding is not easy for high-speed (HS) AFM because the holder that should have a small mass has to be able to clamp the cantilever chip firmly without deteriorating the Z-scanner’s fast performance, and because repeated exchange of cantilever chips should not damage the Z-scanner. This is one of the reasons that tip-scan HS-AFM has not been established, despite its advantages over sample stage-scan HS-AFM. Here, we present a novel method ofmore » cantilever chip holding which meets all conditions required for tip-scan HS-AFM. The superior performance of this novel chip holding mechanism is demonstrated by imaging of the α{sub 3}β{sub 3} subcomplex of F{sub 1}-ATPase in dynamic action at ∼7 frames/s.« less

  3. Integrated Blood Barcode Chips

    PubMed Central

    Fan, Rong; Vermesh, Ophir; Srivastava, Alok; Yen, Brian K.H.; Qin, Lidong; Ahmad, Habib; Kwong, Gabriel A.; Liu, Chao-Chao; Gould, Juliane; Hood, Leroy; Heath, James R.

    2008-01-01

    Blood comprises the largest version of the human proteome1. Changes of plasma protein profiles can reflect physiological or pathological conditions associated with many human diseases, making blood the most important fluid for clinical diagnostics2-4. Nevertheless, only a handful of plasma proteins are utilized in routine clinical tests. This is due to a host of reasons, including the intrinsic complexity of the plasma proteome1, the heterogeneity of human diseases and the fast kinetics associated with protein degradation in sampled blood5. Simple technologies that can sensitively sample large numbers of proteins over broad concentration ranges, from small amounts of blood, and within minutes of sample collection, would assist in solving these problems. Herein, we report on an integrated microfluidic system, called the Integrated Blood Barcode Chip (IBBC). It enables on-chip blood separation and the rapid measurement of a panel of plasma proteins from small quantities of blood samples including a fingerprick of whole blood. This platform holds potential for inexpensive, non-invasive, and informative clinical diagnoses, particularly, for point-of-care. PMID:19029914

  4. High-contrast X-ray micro-tomography of low attenuation samples using large area hybrid semiconductor pixel detector array of 10 × 5 Timepix chips

    NASA Astrophysics Data System (ADS)

    Karch, J.; Krejci, F.; Bartl, B.; Dudak, J.; Kuba, J.; Kvacek, J.; Zemlicka, J.

    2016-01-01

    State-of-the-art hybrid pixel semiconductor detectors provide excellent imaging properties such as unlimited dynamic range, high spatial resolution, high frame rate and energy sensitivity. Nevertheless, a limitation in the use of these devices for imaging has been the small sensitive area of a few square centimetres. In the field of microtomography we make use of a large area pixel detector assembled from 50 Timepix edgeless chips providing fully sensitive area of 14.3 × 7.15 cm2. We have successfully demonstrated that the enlargement of the sensitive area enables high-quality tomographic measurements of whole objects with high geometrical magnification without any significant degradation in resulting reconstructions related to the chip tilling and edgeless sensor technology properties. The technique of micro-tomography with the newly developed large area detector is applied for samples formed by low attenuation, low contrast materials such a seed from Phacelia tanacetifolia, a charcoalified wood sample and a beeswax seal sample.

  5. Using the developed cross-flow filtration chip for collecting blood plasma under high flow rate condition and applying the immunoglobulin E detection

    NASA Astrophysics Data System (ADS)

    Yeh, Chia-Hsien; Hung, Chia-Wei; Wu, Chun-Han; Lin, Yu-Cheng

    2014-09-01

    This paper presents a cross-flow filtration chip for separating blood cells (white blood cells, red blood cells, and platelets) and obtaining blood plasma from human blood. Our strategy is to flow the sample solution in parallel to the membrane, which can generate a parallel shear stress to remove the clogging microparticles on the membrane, so the pure sample solution is obtained in the reservoir. The cross-flow filtration chip includes a cross-flow layer, a Ni-Pd alloy micro-porous membrane, and a reservoir layer. The three layers are packaged in a polymethylmethacrylate (PMMA) frame to create the cross-flow filtration chip. Various dilutions of the blood sample (original, 2 × , 3 × , 5 × , and 10×), pore sizes with different diameters (1 µm, 2 µm, 4 µm, 7 µm, and 10 µm), and different flow rates (1 mL/min, 3 mL/min, 5 mL/min, 7 mL/min, and 10 mL/min) are tested to determine their effects on filtration percentage. The best filtration percentage is 96.2% when the dilution of the blood sample is 10 × , the diameter of pore size of a Ni-Pd alloy micro-porous membrane is 2 µm, and the flow rate is 10 mL/min. Finally, for the clinical tests of the immunoglobulin E (IgE) concentration, the cross-flow filtration chip is used to filter the blood of the allergy patients to obtain the blood plasma. This filtered blood plasma is compared with that obtained using the conventional centrifugation based on the enzyme-linked immunosorbent assay. The results reveal that these two blood separation methods have similar detection trends. The proposed filtration chip has the advantages of low cost, short filtration time, and easy operation and thus can be applied to the separation of microparticles, cells, bacteria, and blood.

  6. Electrochemical detection of methylated DNA on a microfluidic chip with nanoelectrokinetic pre-concentration.

    PubMed

    Hong, Sung A; Kim, Yong-June; Kim, Sung Jae; Yang, Sung

    2018-06-01

    DNA methylation is considered to be a promising marker for the early diagnosis and prognosis of cancer. However, direct detection of the methylated DNAs in clinically relevant samples is still challenging because of its extremely low concentration (~fM). Here, an integrated microfluidic chip is reported, which is capable of pre-concentrating the methylated DNAs using ion concentration polarization (ICP) and electrochemically detecting the pre-concentrated DNAs on a single chip. The proposed chip is the first demonstration of an electrochemical detection of both level and concentration of the methylated DNAs by integrating a DNA pre-concentration unit without gene amplification. Using the proposed chip, 500 fM to 500 nM of methylated DNAs is pre-concentrated by almost 100-fold in 10 min, resulting in a drastic improvement of the electrochemical detection threshold down to the fM level. The proposed chip is able to measure not only the DNA concentration, but also the level of methylation using human urine sample by performing a consecutive electrochemical sensing on a chip. For clinical application, the level as well as the concentration of methylation of glutathione-S transferase-P1 (GSTP1) and EGF-containing fibulin-like extracellular matrix protein 1 (EFEMP1), which are known to be closely associated with prostate cancer diagnosis, are electrochemically detected in human urine spiked with these genes. The developed chip shows a limit of detection (LoD) of 7.9 pM for GSTP1 and 11.8 pM for EFEMP1 and is able to detect the level of methylation in a wide range from 10% to 100% with the concentration variation from 50 pM to 500 nM. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Effect of Vacuum Frying on Changes in Quality Attributes of Jackfruit (Artocarpus heterophyllus) Bulb Slices.

    PubMed

    Maity, Tanushree; Bawa, A S; Raju, P S

    2014-01-01

    The effect of frying temperatures and durations on the quality of vacuum fried jackfruit (JF) chips was evaluated. Moisture content and breaking force of JF chips decreased with increase in frying temperature and time during vacuum frying whereas the oil content increased. The frying time for JF chips was found to be 30, 25, and 20 minutes at 80, 90, and 100°C, respectively. JF chips fried at higher temperature resulted in maximum shrinkage (48%). The lightness in terms of hunter L (*) value decreased significantly (P < 0.05) during frying. Sensory evaluation showed maximum acceptability for JF chips fried at 90°C for 25 min. Frying under vacuum at lower temperatures was found to retain bioactive compounds such as total phenolics, total flavonoids, and total carotenoids in JF chips. Almost 90% of carotenoids were lost from the samples after 30 min of frying at 100°C.

  8. Analysis of aggregates and binders used for the ODOT chip seal program.

    DOT National Transportation Integrated Search

    2010-11-30

    This project compared the results of laboratory characterization of chip seal aggregate samples for Oklahoma DOT Divisions 1,2,3,5 and 6 with performance data from the Pavement Management System (PMS) database. Binder evaluation was limited to identi...

  9. Tracking Silent Hypersensitivity Reactions to Asparaginase during Leukemia Therapy Using Single-Chip Indirect Plasmonic and Fluorescence Immunosensing.

    PubMed

    Charbonneau, David M; Breault-Turcot, Julien; Sinnett, Daniel; Krajinovic, Maja; Leclerc, Jean-Marie; Masson, Jean-François; Pelletier, Joelle N

    2017-12-22

    Microbial asparaginase is an essential component of chemotherapy for the treatment of childhood acute lymphoblastic leukemia (cALL). Silent hypersensitivity reactions to this microbial enzyme need to be monitored accurately during treatment to avoid adverse effects of the drug and its silent inactivation. Here, we present a dual-response anti-asparaginase sensor that combines indirect SPR and fluorescence on a single chip to perform ELISA-type immunosensing, and correlate measurements with classical ELISA. Analysis of serum samples from children undergoing cALL therapy revealed a clear correlation between single-chip indirect SPR/fluorescence immunosensing and ELISA used in clinical settings (R 2 > 0.9). We also report that the portable SPR/fluorescence system had a better sensitivity than classical ELISA to detect antibodies in clinical samples with low antigenicity. This work demonstrates the reliability of dual sensing for monitoring clinically relevant antibody titers in clinical serum samples.

  10. Detection of retromer assembly in Plasmodium falciparum by immunosensing coupled to Surface Plasmon Resonance.

    PubMed

    Iqbal, Mohd Shameel; Siddiqui, Asim Azhar; Banerjee, Chinmoy; Nag, Shiladitya; Mazumder, Somnath; De, Rudranil; Saha, Shubhra Jyoti; Karri, Suresh Kumar; Bandyopadhyay, Uday

    Retromer complex plays a crucial role in intracellular protein trafficking and is conserved throughout the eukaryotes including malaria parasite, Plasmodium falciparum, where it is partially conserved. The assembly of retromer complex in RBC stages of malarial parasite is extremely difficult to explore because of its complicated physiology, small size, and intra-erythrocytic location. Nonetheless, understanding of retromer assembly may pave new ways for the development of novel antimalarials targeting parasite-specific protein trafficking pathways. Here, we investigated the assembly of retromer complex in P. falciparum, by an immunosensing method through highly sensitive Surface Plasmon Resonance (SPR) technique. After taking leads from the bioinformatics search and literature, different interacting proteins were identified and specific antibodies were raised against them. The sensor chip was prepared by covalently linking antibody specific to one component and the whole cell lysate was passed through it in order to trap the interacting complex. Antibodies raised against other interacting components were used to detect them in the trapped complex on the SPR chip. We were able to detect three different components in the retromer complex trapped by the immobilized antibody specific against a different component on a sensor chip. The assay was reproduced and validated in a different two-component CD74-MIF system in mammalian cells. We, thus, illustrate the assembly of retromer complex in P. falciparum through a bio-sensing approach that combines SPR with immunosensing requiring a very small amount of sample from the native source. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Characterization of aqueous two phase systems by combining lab-on-a-chip technology with robotic liquid handling stations.

    PubMed

    Amrhein, Sven; Schwab, Marie-Luise; Hoffmann, Marc; Hubbuch, Jürgen

    2014-11-07

    Over the last decade, the use of design of experiment approaches in combination with fully automated high throughput (HTP) compatible screenings supported by robotic liquid handling stations (LHS), adequate fast analytics and data processing has been developed in the biopharmaceutical industry into a strategy of high throughput process development (HTPD) resulting in lower experimental effort, sample reduction and an overall higher degree of process optimization. Apart from HTP technologies, lab-on-a-chip technology has experienced an enormous growth in the last years and allows further reduction of sample consumption. A combination of LHS and lab-on-a-chip technology is highly desirable and realized in the present work to characterize aqueous two phase systems with respect to tie lines. In particular, a new high throughput compatible approach for the characterization of aqueous two phase systems regarding tie lines by exploiting differences in phase densities is presented. Densities were measured by a standalone micro fluidic liquid density sensor, which was integrated into a liquid handling station by means of a developed generic Tip2World interface. This combination of liquid handling stations and lab-on-a-chip technology enables fast, fully automated, and highly accurate density measurements. The presented approach was used to determine the phase diagram of ATPSs composed of potassium phosphate (pH 7) and polyethylene glycol (PEG) with a molecular weight of 300, 400, 600 and 1000 Da respectively in the presence and in the absence of 3% (w/w) sodium chloride. Considering the whole ATPS characterization process, two complete ATPSs could be characterized within 24h, including four runs per ATPS for binodal curve determination (less than 45 min/run), and tie line determination (less than 45 min/run for ATPS preparation and 8h for density determination), which can be performed fully automated over night without requiring man power. The presented methodology provides a cost, time and material effective approach for characterization of ATPS phase diagram on base on highly accurate and comprehensive data. By this means the derived data opens the door for a more detailed description of ATPS towards generating mechanistic based models, since molecular approaches such as MD simulations or molecular descriptions along the line of QSAR heavily rely on accurate and comprehensive data. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. In-situ X-ray CT results of damage evolution in L6 ordinary chondrite meteorites

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cuadra, Jefferson A.; Hazeli, Kavan; Ramesh, K. T.

    2016-06-17

    These are slides about in-situ X-ray CT results of damage evolution in L6 ordinary chondrite meteorites. The following topics are covered: mechanical and thermal damage characterization, list of Grosvenor Mountain (GRO) meteorite samples, in-situ x-ray compression test setup, GRO-chipped reference at 0 N - existing cracks, GRO-chipped loaded at 1580 N, in-situ x-ray thermal fatigue test setup, GRO-B14 room temperature reference, GRO-B14 Cycle 47 at 200°C, GRO-B14 Cycle 47 at room temperature, conclusions from qualitative analysis, future work and next steps. Conclusions are the following: Both GRO-Chipped and GRO-B14 had existing voids and cracks within the volume. These sites withmore » existing damage were selected for CT images from mechanically and thermally loaded scans since they are prone to damage initiation. The GRO-Chipped sample was loaded to 1580 N which resulted in a 14% compressive engineering strain, calculated using LVDT. Based on the CT cross sectional images, the GRO-B14 sample at 200°C has a thermal expansion of approximately 96 μm in height (i.e. ~1.6% engineering strain).« less

  13. Microchip systems for immunoassay: an integrated immunoreactor with electrophoretic separation for serum theophylline determination.

    PubMed

    Chiem, N H; Harrison, D J

    1998-03-01

    A glass microchip is described in which reagents and serum samples for competitive immunoassay of serum theophylline can be mixed, reacted, separated, and analyzed. The device functions as an automated microfluidic immunoassay system, creating a lab-on-a-chip. Electroosmotic pumping was used to control first the mixing of 50-fold-diluted serum sample with labeled theophylline tracer in a 1:1 ratio, followed by 1:1 mixing and reaction with anti-theophylline antibody. The 51-nL on-chip mixer gave the same concentration as dilution performed off-chip, within 3%. A 100-pL plug of the reacted solution was then injected into an electrophoresis separation channel integrated within the same chip. Measurements of free and bound tracer by fluorescence detection gave linear calibration curves of signal vs log[theophylline] between 0 and 40 mg/L, with a slope of 0.52 +/- 0.03 and an intercept of -0.04 +/- 0.04 after a 90-s reaction time. A detection limit of 0.26 mg/L in serum (expressed before the dilution step, actual concentration of 1.3 micrograms/L at the detector) was obtained. Recovery values were 107% +/- 8% for 15 mg/L serum samples.

  14. Microfluidic Devices for Forensic DNA Analysis: A Review

    PubMed Central

    Bruijns, Brigitte; van Asten, Arian; Tiggelaar, Roald; Gardeniers, Han

    2016-01-01

    Microfluidic devices may offer various advantages for forensic DNA analysis, such as reduced risk of contamination, shorter analysis time and direct application at the crime scene. Microfluidic chip technology has already proven to be functional and effective within medical applications, such as for point-of-care use. In the forensic field, one may expect microfluidic technology to become particularly relevant for the analysis of biological traces containing human DNA. This would require a number of consecutive steps, including sample work up, DNA amplification and detection, as well as secure storage of the sample. This article provides an extensive overview of microfluidic devices for cell lysis, DNA extraction and purification, DNA amplification and detection and analysis techniques for DNA. Topics to be discussed are polymerase chain reaction (PCR) on-chip, digital PCR (dPCR), isothermal amplification on-chip, chip materials, integrated devices and commercially available techniques. A critical overview of the opportunities and challenges of the use of chips is discussed, and developments made in forensic DNA analysis over the past 10–20 years with microfluidic systems are described. Areas in which further research is needed are indicated in a future outlook. PMID:27527231

  15. Parents and the V-Chip: A Kaiser Family Foundation Survey. How Parents Feel (and What They Know) about TV, the V-Chip, and the TV Ratings System. Toplines.

    ERIC Educational Resources Information Center

    1999

    This report presents the data from a national, random sample survey designed by the Kaiser Family Foundation and Princeton Survey Research Associates (PSRA) and conducted by PSRA by telephone between April 6-26, 1999, when 1001 parents of children ages 2-17 were interviewed about their opinions of television, the V-Chip, and the TV ratings system.…

  16. Development of Criteria for the Use of Asphalt-Rubber as a Stress-Absorbing Membrane Interlayer (SAMI).

    DTIC Science & Technology

    1983-12-01

    membrane included " three asphalt-rubber distributor trucks " one asphalt-rubber mixer • one kerosene tanker " one chip spreader " three dump trucks...was recorded with the temperature. 4. The asphalt-rubber was pumped into a spreader truck. The truck temperature, sample temperature, and viscosity...not become clogged. The chip-spreading operation immnedi- ately followed the distributor truck. The chip spreader was immediately fol- lowed by the

  17. Microfabrication of three-dimensional filters for liposome extrusion

    NASA Astrophysics Data System (ADS)

    Baldacchini, Tommaso; Nuñez, Vicente; LaFratta, Christopher N.; Grech, Joseph S.; Vullev, Valentine I.; Zadoyan, Ruben

    2015-03-01

    Liposomes play a relevant role in the biomedical field of drug delivery. The ability of these lipid vesicles to encapsulate and transport a variety of bioactive molecules has fostered their use in several therapeutic applications, from cancer treatments to the administration of drugs with antiviral activities. Size and uniformity are key parameters to take into consideration when preparing liposomes; these factors greatly influence their effectiveness in both in vitro and in vivo experiments. A popular technique employed to achieve the optimal liposome dimension (around 100 nm in diameter) and uniform size distribution is repetitive extrusion through a polycarbonate filter. We investigated two femtosecond laser direct writing techniques for the fabrication of three-dimensional filters within a microfluidics chip for liposomes extrusion. The miniaturization of the extrusion process in a microfluidic system is the first step toward a complete solution for lab-on-a-chip preparation of liposomes from vesicles self-assembly to optical characterization.

  18. CMOS active pixel sensor type imaging system on a chip

    NASA Technical Reports Server (NTRS)

    Fossum, Eric R. (Inventor); Nixon, Robert (Inventor)

    2011-01-01

    A single chip camera which includes an .[.intergrated.]. .Iadd.integrated .Iaddend.image acquisition portion and control portion and which has double sampling/noise reduction capabilities thereon. Part of the .[.intergrated.]. .Iadd.integrated .Iaddend.structure reduces the noise that is picked up during imaging.

  19. Characteristics and Frequency of Chipping Effects in Near-Contact Gunshot Wounds.

    PubMed

    Amadasi, Alberto; Mazzarelli, Debora; Merli, Daniele; Brandone, Alberto; Cattaneo, Cristina

    2017-05-01

    The presence of "chipping" or "flaking" around the edges of gunshot entry wounds has been described among the characteristics of gunshot wounds in bone. In this study, the real frequency of such a peculiar feature was investigated. The presence of "chipping" was assessed on 22 gunshot wounds fired at a near-contact range on bovine ribs with 9-mm bullets. As controls, five samples were shot with a 3 cm range, and five from 40 cm. In 77% of cases shot at near-contact range, a detachment of small fragments of the upper layers of bone was detected, mainly with a circumferential disposition, whereas this feature was lacking in control samples. The study demonstrated the frequency of "chipping" and that it may probably be due to a combined ballistic effect of impact of the bullet itself and expansion of gases. It may be thus considered indicative of close-range shots. © 2016 American Academy of Forensic Sciences.

  20. Design and status of the RF-digitizer integrated circuit

    NASA Technical Reports Server (NTRS)

    Rayhrer, B.; Lam, B.; Young, L. E.; Srinivasan, J. M.; Thomas, J. B.

    1991-01-01

    An integrated circuit currently under development samples a bandpass-limited signal at a radio frequency in quadrature and then performs a simple sum-and-dump operation in order to filter and lower the rate of the samples. Downconversion to baseband is carried out by the sampling step itself through the aliasing effect of an appropriately selected subharmonic sampling frequency. Two complete RF digitizer circuits with these functions will be implemented with analog and digital elements on one GaAs substrate. An input signal, with a carrier frequency as high as 8 GHz, can be sampled at a rate as high as 600 Msamples/sec for each quadrature component. The initial version of the chip will sign-sample (1-bit) the input RF signal. The chip will contain a synthesizer to generate a sample frequency that is a selectable integer multiple of an input reference frequency. In addition to the usual advantages of compactness and reliability associated with integrated circuits, the single chip will replace several steps required by standard analog downconversion. Furthermore, when a very high initial sample rate is selected, the presampling analog filters can be given very large bandwidths, thereby greatly reducing phase and delay instabilities typically introduced by such filters, as well as phase and delay variation due to Doppler changes.

  1. Molecular beacon modified sensor chips for oligonucleotide detection with optical readout.

    PubMed

    Su, Qiang; Wesner, Daniel; Schönherr, Holger; Nöll, Gilbert

    2014-12-02

    Three different surface bound molecular beacons (MBs) were investigated using surface plasmon fluorescence spectroscopy (SPFS) as an optical readout technique. While MB1 and MB2, both consisting of 36 bases, differed only in the length of the linker for surface attachment, the significantly longer MB3, consisting of 56 bases, comprised an entirely different sequence. For sensor chip preparation, the MBs were chemisorbed on gold via thiol anchors together with different thiol spacers. The influence of important parameters, such as the length of the MBs, the length of the linker between the MBs and the gold surface, the length and nature of the thiol spacers, and the ratio between the MBs and the thiol spacers was studied. After hybridization with the target, the fluorophore of the longer MB3 was oriented close to the surface, and the shorter MBs were standing more or less upright, leading to a larger increase in fluorescence intensity. Fluorescence microscopy revealed a homogeneous distribution of the MBs on the surface. The sensor chips could be used for simple and fast detection of target molecules with a limit of detection in the larger picomolar range. The response time was between 5 and 20 min. Furthermore, it was possible to distinguish between fully complementary and singly mismatched targets. While rinsing with buffer solution after hybridization with target did not result in any signal decrease, complete dehybridization could be carried out by intense rinsing with pure water. The MB modified sensor chips could be prepared in a repeatable manner and reused many times without significant decrease in performance.

  2. Ultrathin Polymer Membranes with Patterned, Micrometric Pores for Organs-on-Chips.

    PubMed

    Pensabene, Virginia; Costa, Lino; Terekhov, Alexander Y; Gnecco, Juan S; Wikswo, John P; Hofmeister, William H

    2016-08-31

    The basal lamina or basement membrane (BM) is a key physiological system that participates in physicochemical signaling between tissue types. Its formation and function are essential in tissue maintenance, growth, angiogenesis, disease progression, and immunology. In vitro models of the BM (e.g., Boyden and transwell chambers) are common in cell biology and lab-on-a-chip devices where cells require apical and basolateral polarization. Extravasation, intravasation, membrane transport of chemokines, cytokines, chemotaxis of cells, and other key functions are routinely studied in these models. The goal of the present study was to integrate a semipermeable ultrathin polymer membrane with precisely positioned pores of 2 μm diameter in a microfluidic device with apical and basolateral chambers. We selected poly(l-lactic acid) (PLLA), a transparent biocompatible polymer, to prepare the semipermeable ultrathin membranes. The pores were generated by pattern transfer using a three-step method coupling femtosecond laser machining, polymer replication, and spin coating. Each step of the fabrication process was characterized by scanning electron microscopy to investigate reliability of the process and fidelity of pattern transfer. In order to evaluate the compatibility of the fabrication method with organs-on-a-chip technology, porous PLLA membranes were embedded in polydimethylsiloxane (PDMS) microfluidic devices and used to grow human umbilical vein endothelial cells (HUVECS) on top of the membrane with perfusion through the basolateral chamber. Viability of cells, optical transparency of membranes and strong adhesion of PLLA to PDMS were observed, thus confirming the suitability of the prepared membranes for use in organs-on-a-chip devices.

  3. A poly(dimethylsiloxane) microfluidic sheet reversibly adhered on a glass plate for creation of emulsion droplets for droplet digital PCR.

    PubMed

    Nakashoji, Yuta; Tanaka, Hironari; Tsukagoshi, Kazuhiko; Hashimoto, Masahiko

    2017-01-01

    A PDMS microfluidic chip with T-junction channel geometry, two inlet reservoirs, and one outlet reservoir was reversibly adhered on a glass plate through the viscoelastic properties of PDMS. This formed a detachable microfluidic device for creation of water-in-oil emulsion droplets that were used as discrete reaction compartments for the droplet digital PCR. The PDMS/glass device could continuously produce monodisperse droplets without leakage of fluids using a vacuum-driven autonomous micropumping method. This droplet preparation technique only required evacuation of air dissolved in the PDMS before loading of oil and aqueous phases into separate inlet reservoirs. Degassing of the PDMS chip at approximately 300 Pa for 1.5 h in a vacuum desiccator gave 40 000 droplets in 80 min, which corresponded to a generation frequency of up to nine droplets per second. Over multiple runs the droplet creation was very reproducible, and the size reproducibility of generated droplets (polydispersity of up to 4.1%) was comparable to that acquired using other microfluidic droplet preparation techniques. Because the PDMS chip can be peeled off the glass plate, blocked channels can easily be fixed when they arise, and this extends the lifetime of the chip. Single DNA molecules partitioned into the droplets were successfully amplified by PCR. In addition, the droplet digital PCR platform allowed absolute quantification of low copy numbers of target DNA, and was robust against instrumental variance. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Electrochemical microfluidic chip based on molecular imprinting technique applied for therapeutic drug monitoring.

    PubMed

    Liu, Jiang; Zhang, Yu; Jiang, Min; Tian, Liping; Sun, Shiguo; Zhao, Na; Zhao, Feilang; Li, Yingchun

    2017-05-15

    In this work, a novel electrochemical detection platform was established by integrating molecularly imprinting technique with microfluidic chip and applied for trace measurement of three therapeutic drugs. The chip foundation is acrylic panel with designed grooves. In the detection cell of the chip, a Pt wire is used as the counter electrode and reference electrode, and a Au-Ag alloy microwire (NPAMW) with 3D nanoporous surface modified with electro-polymerized molecularly imprinted polymer (MIP) film as the working electrode. Detailed characterization of the chip and the working electrode was performed, and the properties were explored by cyclic voltammetry and electrochemical impedance spectroscopy. Two methods, respectively based on electrochemical catalysis and MIP/gate effect were employed for detecting warfarin sodium by using the prepared chip. The linearity of electrochemical catalysis method was in the range of 5×10 -6 -4×10 -4 M, which fails to meet clinical testing demand. By contrast, the linearity of gate effect was 2×10 -11 -4×10 -9 M with remarkably low detection limit of 8×10 -12 M (S/N=3), which is able to satisfy clinical assay. Then the system was applied for 24-h monitoring of drug concentration in plasma after administration of warfarin sodium in rabbit, and the corresponding pharmacokinetic parameters were obtained. In addition, the microfluidic chip was successfully adopted to analyze cyclophosphamide and carbamazepine, implying its good versatile ability. It is expected that this novel electrochemical microfluidic chip can act as a promising format for point-of-care testing via monitoring different analytes sensitively and conveniently. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Microfluidic "Pouch" Chips for Immunoassays and Nucleic Acid Amplification Tests.

    PubMed

    Mauk, Michael G; Liu, Changchun; Qiu, Xianbo; Chen, Dafeng; Song, Jinzhao; Bau, Haim H

    2017-01-01

    Microfluidic cassettes ("chips") for processing and analysis of clinical specimens and other sample types facilitate point-of-care (POC) immunoassays and nucleic acid based amplification tests. These single-use test chips can be self-contained and made amenable to autonomous operation-reducing or eliminating supporting instrumentation-by incorporating laminated, pliable "pouch" and membrane structures for fluid storage, pumping, mixing, and flow control. Materials and methods for integrating flexible pouch compartments and diaphragm valves into hard plastic (e.g., acrylic and polycarbonate) microfluidic "chips" for reagent storage, fluid actuation, and flow control are described. We review several versions of these pouch chips for immunoassay and nucleic acid amplification tests, and describe related fabrication techniques. These protocols thus offer a "toolbox" of methods for storage, pumping, and flow control functions in microfluidic devices.

  6. Rapid analysis of protein interactions: On-chip micropurification of recombinant protein expressed in Esherichia coli.

    PubMed

    Natsume, Tohru; Taoka, Masato; Manki, Hiroshi; Kume, Shouen; Isobe, Toshiaki; Mikoshiba, Katsuhiko

    2002-09-01

    We describe a rapid analysis of interactions between antibodies and a recombinant protein present in total cell lysates. Using a surface plasmon resonance biosensor, a low concentration of glutathione-S-transferase (GST) fused protein expressed in small scale Esherichia coli culture was purified on an anti-GST antibody immobilized sensor chip. The 'on-chip purification' was verified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry by measuring the molecular masses of recombinant proteins purified on the sensor chip. The specific binding of monoclonal antibodies for the on-chip micropurified recombinant proteins can then be monitored, thus enabling kinetic analysis and epitope mapping of the bound antibodies. This approach reduced time, resources and sample consumption by avoiding conventional steps related to concentration and purification.

  7. Portable Apparatus for Electrochemical Sensing of Ethylene

    NASA Technical Reports Server (NTRS)

    Manoukian, Mourad; Tempelman, Linda A.; Forchione, John; Krebs, W. Michael; Schmitt, Edwin W.

    2007-01-01

    A small, lightweight, portable apparatus based on an electrochemical sensing principle has been developed for monitoring low concentrations of ethylene in air. Ethylene has long been known to be produced by plants and to stimulate the growth and other aspects of the development of plants (including, notably, ripening of fruits and vegetables), even at concentrations as low as tens of parts per billion (ppb). The effects are magnified in plant-growth and -storage chambers wherein ethylene can accumulate. There is increasing recognition in agriculture and related industries that it is desirable to monitor and control ethylene concentrations in order to optimize the growth, storage, and ripening of plant products. Hence, there are numerous potential uses for the present apparatus in conjunction with equipment for controlling ethylene concentrations. The ethylene sensor is of a thick-film type with a design optimized for a low detection limit. The sensor includes a noble metal sensing electrode on a chip and a hydrated solid-electrolyte membrane that is held in contact with the chip. Also located on the sensor chip are a counter electrode and a reference electrode. The sensing electrode is held at a fixed potential versus the reference electrode. Detection takes place at active-triple-point areas where the sensing electrode, electrolyte, and sample gas meet. These areas are formed by cutting openings in the electrolyte membrane. The electrode current generated from electrochemical oxidation of ethylene at the active triple points is proportional to the concentration of ethylene. An additional film of the solid-electrolyte membrane material is deposited on the sensing electrode to increase the effective triple-point areas and thereby enhance the detection signal. The sensor chip is placed in a holder that is part of a polycarbonate housing. When fully assembled, the housing holds the solid-electrolyte membrane in contact with the chip (see figure). The housing includes a water reservoir for keeping the solid-electrolyte membrane hydrated. The housing also includes flow channels for circulating a sample stream of air over the chip: ethylene is brought to the sensing surface predominately by convection in this sample stream. The sample stream is generated by a built-in sampling pump. The forced circulation of sample air contributes to the attainment of a low detection limit.

  8. On-Chip Spyhole Nanoelectrospray Ionization Mass Spectrometry for Sensitive Biomarker Detection in Small Volumes

    NASA Astrophysics Data System (ADS)

    Zhong, Xiaoqin; Qiao, Liang; Stauffer, Géraldine; Liu, Baohong; Girault, Hubert H.

    2018-03-01

    A polyimide microfluidic chip with a microhole emitter (Ø 10-12 μm) created on top of a microchannel by scanning laser ablation has been designed for nanoelectrospray ionization (spyhole-nanoESI) to couple microfluidics with mass spectrometry. The spyhole-nanoESI showed higher sensitivity compared to standard ESI and microESI from the end of the microchannel. The limits of detection (LOD) for peptide with the spyhole-nanoESI MS reached 50 pM, which was 600 times lower than that with standard ESI. The present microchip emitter allows the analysis of small volumes of samples. As an example, a small cell lung cancer biomarker, neuron-specific enolase (NSE), was detected by monitoring the transition of its unique peptide with the spyhole-nanoESI MS/MS. NSE at 0.2 nM could be well identified with a signal to noise ratio (S/N) of 50, and thereby its LOD was estimated to be 12 pM. The potential application of the spyhole-nanoESI MS/MS in cancer diagnosis was further demonstrated with the successful detection of 2 nM NSE from 1 μL of human serum. Before the detection, the serum sample spiked with NSE was first depleted with immune spin column, then desalted by centrifugal filter device, and finally digested by trypsin, without any other complicated preparation steps. The concentration matched the real condition of clinical samples. In addition, the microchips can be disposable to avoid any cross contamination. The present technique provides a highly efficient way to couple microfluidics with MS, which brings additional values to various microfluidics and MS-based analysis.

  9. Application of the CometChip platform to assess DNA damage in field-collected blood samples from turtles.

    PubMed

    Sykora, Peter; Chiari, Ylenia; Heaton, Andrew; Moreno, Nickolas; Glaberman, Scott; Sobol, Robert W

    2018-05-01

    DNA damage has been linked to genomic instability and the progressive breakdown of cellular and organismal homeostasis, leading to the onset of disease and reduced longevity. Insults to DNA from endogenous sources include base deamination, base hydrolysis, base alkylation, and metabolism-induced oxidative damage that can lead to single-strand and double-strand DNA breaks. Alternatively, exposure to environmental pollutants, radiation or ultra-violet light, can also contribute to exogenously derived DNA damage. We previously validated a novel, high through-put approach to measure levels of DNA damage in cultured mammalian cells. This new CometChip Platform builds on the classical single cell gel electrophoresis or comet methodology used extensively in environmental toxicology and molecular biology. We asked whether the CometChip Platform could be used to measure DNA damage in samples derived from environmental field studies. To this end, we determined that nucleated erythrocytes from multiple species of turtle could be successfully evaluated in the CometChip Platform to quantify levels of DNA damage. In total, we compared levels of DNA damage in 40 animals from two species: the box turtle (Terrapene carolina) and the red-eared slider (Trachemys scripta elegans). Endogenous levels of DNA damage were identical between the two species, yet we did discover some sex-linked differences and changes in DNA damage accumulation. Based on these results, we confirm that the CometChip Platform allows for the measurement of DNA damage in a large number of samples quickly and accurately, and is particularly adaptable to environmental studies using field-collected samples. Environ. Mol. Mutagen. 59:322-333, 2018. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

  10. A compressive-sensing Fourier-transform on-chip Raman spectrometer

    NASA Astrophysics Data System (ADS)

    Podmore, Hugh; Scott, Alan; Lee, Regina

    2018-02-01

    We demonstrate a novel compressive sensing Fourier-transform spectrometer (FTS) for snapshot Raman spectroscopy in a compact format. The on-chip FTS consists of a set of planar-waveguide Mach-Zehnder interferometers (MZIs) arrayed on a photonic chip, effecting a discrete Fourier-transform of the input spectrum. Incoherence between the sampling domain (time), and the spectral domain (frequency) permits compressive sensing retrieval using undersampled interferograms for sparse spectra such as Raman emission. In our fabricated device we retain our chosen bandwidth and resolution while reducing the number of MZIs, e.g. the size of the interferogram, to 1/4th critical sampling. This architecture simultaneously reduces chip footprint and concentrates the interferogram in fewer pixels to improve the signal to noise ratio. Our device collects interferogram samples simultaneously, therefore a time-gated detector may be used to separate Raman peaks from sample fluorescence. A challenge for FTS waveguide spectrometers is to achieve multi-aperture high throughput broadband coupling to a large number of single-mode waveguides. A multi-aperture design allows one to increase the bandwidth and spectral resolution without sacrificing optical throughput. In this device, multi-aperture coupling is achieved using an array of microlenses bonded to the surface of the chip, and aligned with a grid of vertically illuminated waveguide apertures. The microlens array accepts a collimated beam with near 100% fill-factor, and the resulting spherical wavefronts are coupled into the single-mode waveguides using 45& mirrors etched into the waveguide layer via focused ion-beam (FIB). The interferogram from the waveguide outputs is imaged using a CCD, and inverted via l1-norm minimization to correctly retrieve a sparse input spectrum.

  11. Future lab-on-a-chip technologies for interrogating individual molecules.

    PubMed

    Craighead, Harold

    2006-07-27

    Advances in technology have allowed chemical sampling with high spatial resolution and the manipulation and measurement of individual molecules. Adaptation of these approaches to lab-on-a-chip formats is providing a new class of research tools for the investigation of biochemistry and life processes.

  12. Multi-GHz Synchronous Waveform Acquisition With Real-Time Pattern-Matching Trigger Generation

    NASA Astrophysics Data System (ADS)

    Kleinfelder, Stuart A.; Chiang, Shiuh-hua Wood; Huang, Wei

    2013-10-01

    A transient waveform capture and digitization circuit with continuous synchronous 2-GHz sampling capability and real-time programmable windowed trigger generation has been fabricated and tested. Designed in 0.25 μm CMOS, the digitizer contains a circular array of 128 sample and hold circuits for continuous sample acquisition, and attains 2-GHz sample speeds with over 800-MHz analog bandwidth. Sample clock generation is synchronous, combining a phase-locked loop for high-speed clock generation and a high-speed fully-differential shift register for distributing clocks to all 128 sample circuits. Using two comparators per sample, the sampled voltage levels are compared against two reference levels, a high threshold and a low threshold, that are set via per-comparator digital to analog converters (DACs). The 256 per-comparator 5-bit DACs compensate for comparator offsets and allow for fine reference level adjustment. The comparator results are matched in 8-sample-wide windows against up to 72 programmable patterns in real time using an on-chip programmable logic array. Each 8-sample trigger window is equivalent to 4 ns of acquisition, overlapped sample by sample in a circular fashion through the entire 128-sample array. The 72 pattern-matching trigger criteria can be programmed to be any combination of High-above the high threshold, Low-below the low threshold, Middle-between the two thresholds, or “Don't Care”-any state is accepted. A trigger pattern of “HLHLHLHL,” for example, watches for a waveform that is oscillating at about 1 GHz given the 2-GHz sample rate. A trigger is flagged in under 20 ns if there is a match, after which sampling is stopped, and on-chip digitization can proceed via 128 parallel 10-bit converters, or off-chip conversion can proceed via an analog readout. The chip exceeds 11 bits of dynamic range, nets over 800-MHz -3-dB bandwidth in a realistic system, and jitter in the PLL-based sampling clock has been measured to be about 1 part per million, RMS.

  13. Suitability of shredded tyres as a substitute for a landfill leachate collection medium.

    PubMed

    Park, Jae K; Edil, Tuncer B; Kim, Jae Y; Huh, Mock; Lee, Sung Ho; Lee, Jung Jun

    2003-06-01

    A series of tests were conducted to investigate the fate of heavy metals and gasoline components in a simulated landfill, consisting of a 30 cm thick clay liner and a leachate collection layer containing tyres as well as in two test cells installed in a landfill. Arsenic, selenium, mercury, barium, and lead concentrations were lower while zinc concentration was higher in the tank containing tyre-chips than the tank without tyre-chips. When samples were filtered, however, concentrations of zinc as well as other inorganics were lower in the tank containing tyre-chips, indicating that metals in the leachate exposed to tyre-chips travel more slowly in a subsurface environment due to filtering effect. In a test cell study, arsenic, cobalt, lead and nickel concentrations were lower in the cell containing tyre-chips than in the cell without tyre-chips, except iron and zinc. Both tests indicate that some inorganic contaminants are sorbed to tyre-chips. Gasoline components were also significantly sorbed by tyre-chips in field cell tests. Although tyre-chips are known to leach organic and inorganic contaminants, concentrations in field conditions will be lower than the reported experimental results since the tests were performed under worst-case scenarios. If tyre-chips are used in areas where contamination levels are high, then they can be used as a sorbent for environmental clean-up.

  14. The Chaperone-assisted E3 Ligase C Terminus of Hsc70-interacting Protein (CHIP) Targets PTEN for Proteasomal Degradation*

    PubMed Central

    Ahmed, Syed Feroj; Deb, Satamita; Paul, Indranil; Chatterjee, Anirban; Mandal, Tapashi; Chatterjee, Uttara; Ghosh, Mrinal K.

    2012-01-01

    The tumor suppressor, PTEN is key to the regulation of diverse cellular processes, making it a prime candidate to be tightly regulated. The PTEN level is controlled in a major way by E3 ligase-mediated degradation through the Ubiquitin-Proteasome System (UPS). Nedd 4-1, XIAP, and WWP2 have been shown to maintain PTEN turnover. Here, we report that CHIP, the chaperone-associated E3 ligase, induces ubiquitination and regulates the proteasomal turnover of PTEN. It was apparent from our findings that PTEN transiently associates with the molecular chaperones and thereby gets diverted to the degradation pathway through its interaction with CHIP. The TPR domain of CHIP and parts of the N-terminal domain of PTEN are required for their interaction. Overexpression of CHIP leads to elevated ubiquitination and a shortened half-life of endogenous PTEN. On the other hand, depletion of endogenous CHIP stabilizes PTEN. CHIP is also shown to regulate PTEN-dependent transcription presumably through its down-regulation. PTEN shared an inverse correlation with CHIP in human prostate cancer patient samples, thereby triggering the prospects of a more complex mode of PTEN regulation in cancer. PMID:22427670

  15. Chip-based device for parallel sorting, amplification, detection, and identification of nucleic acid subsequences

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Beer, Neil Reginald; Colston, Jr, Billy W.

    An apparatus for chip-based sorting, amplification, detection, and identification of a sample having a planar substrate. The planar substrate is divided into cells. The cells are arranged on the planar substrate in rows and columns. Electrodes are located in the cells. A micro-reactor maker produces micro-reactors containing the sample. The micro-reactor maker is positioned to deliver the micro-reactors to the planar substrate. A microprocessor is connected to the electrodes for manipulating the micro-reactors on the planar substrate. A detector is positioned to interrogate the sample contained in the micro-reactors.

  16. Wide-field lensless fluorescent microscopy using a tapered fiber-optic faceplate on a chip.

    PubMed

    Coskun, Ahmet F; Sencan, Ikbal; Su, Ting-Wei; Ozcan, Aydogan

    2011-09-07

    We demonstrate lensless fluorescent microscopy over a large field-of-view of ~60 mm(2) with a spatial resolution of <4 µm. In this on-chip fluorescent imaging modality, the samples are placed on a fiber-optic faceplate that is tapered such that the density of the fiber-optic waveguides on the top facet is >5 fold larger than the bottom one. Placed on this tapered faceplate, the fluorescent samples are pumped from the side through a glass hemisphere interface. After excitation of the samples, the pump light is rejected through total internal reflection that occurs at the bottom facet of the sample substrate. The fluorescent emission from the sample is then collected by the smaller end of the tapered faceplate and is delivered to an opto-electronic sensor-array to be digitally sampled. Using a compressive sampling algorithm, we decode these raw lensfree images to validate the resolution (<4 µm) of this on-chip fluorescent imaging platform using microparticles as well as labeled Giardia muris cysts. This wide-field lensfree fluorescent microscopy platform, being compact and high-throughput, might provide a valuable tool especially for cytometry, rare cell analysis (involving large area microfluidic systems) as well as for microarray imaging applications.

  17. Microfluidic high performance liquid chromatography-chip hyphenation to inductively coupled plasma-mass spectrometry.

    PubMed

    Bishop, David P; Blanes, Lucas; Wilson, Alexander B; Wilbanks, Thor; Killeen, Kevin; Grimm, Rudolf; Wenzel, Ross; Major, Derek; Macka, Mirek; Clarke, David; Schmid, Robin; Cole, Nerida; Doble, Philip A

    2017-05-12

    The Agilent Chip Cube Interface is a microfluidic chip-based technology originally designed for nanospray molecular mass spectrometry in which the sample enrichment, nano-column, tubing, connectors and spray tip were integrated into a single biocompatible chip. Here we describe the hyphenation of the Chip Cube Interface to ICP-MS via modification of the standard HPLC chip design and a new total consumption nebuliser suitable for flow rates as low as 300nLmin -1 . The potential of the instrument to eliminate common nanoLC - ICP-MS shortcomings such as leaks, blockages and band-broadening was demonstrated via analysis of cyanocobalamin in equine plasma. The method was linear over three orders of magnitude with an r 2 of 0.9999, the peak area repeatability was 1.9% (n=7), and the detection limit was 14ngmL -1 . This novel configuration of the Chip Cube Interface coupled to ICP-MS is a suitable platform for the analysis of biomolecules associated with trace metals and speciation applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Storage insects on yam chips and their traditional management in Northern Benin.

    PubMed

    Loko, Y L; Dansi, A; Tamo, M; Bokonon-Ganta, A H; Assogba, P; Dansi, M; Vodouhè, R; Akoegninou, A; Sanni, A

    2013-01-01

    Twenty-five villages of Northern Benin were surveyed to identify the constraints of yam chips production, assess the diversity of storage insects on yam chips, and document farmers' perception of their impacts on the stocks and their traditional management practices. Damages due to storage insects (63.9% of responses) and insufficiency of insect-resistant varieties (16.7% of responses) were the major constraints of yam chips production. Twelve insect pest species were identified among which Dinoderus porcellus Lesne (Coleoptera, Bostrichidae) was by far the most important and the most distributed (97.44% of the samples). Three predators (Teretrius nigrescens Lewis, Xylocoris flavipes Reuter, and Alloeocranum biannulipes Montrouzier & Signoret) and one parasitoid (Dinarmus basalis Rondani) all Coleoptera, Bostrichidae were also identified. The most important traditional practices used to control or prevent insect attack in yam chips were documented and the producers' preference criteria for yam cultivars used to produce chips were identified and prioritized. To further promote the production of yam chips, diversification of insect-resistant yam varieties, conception, and use of health-protective natural insecticides and popularization of modern storage structures were proposed.

  19. Storage Insects on Yam Chips and Their Traditional Management in Northern Benin

    PubMed Central

    Loko, Y. L.; Dansi, A.; Tamo, M.; Bokonon-Ganta, A. H.; Assogba, P.; Dansi, M.; Vodouhè, R.; Akoegninou, A.; Sanni, A.

    2013-01-01

    Twenty-five villages of Northern Benin were surveyed to identify the constraints of yam chips production, assess the diversity of storage insects on yam chips, and document farmers' perception of their impacts on the stocks and their traditional management practices. Damages due to storage insects (63.9% of responses) and insufficiency of insect-resistant varieties (16.7% of responses) were the major constraints of yam chips production. Twelve insect pest species were identified among which Dinoderus porcellus Lesne (Coleoptera, Bostrichidae) was by far the most important and the most distributed (97.44% of the samples). Three predators (Teretrius nigrescens Lewis, Xylocoris flavipes Reuter, and Alloeocranum biannulipes Montrouzier & Signoret) and one parasitoid (Dinarmus basalis Rondani) all Coleoptera, Bostrichidae were also identified. The most important traditional practices used to control or prevent insect attack in yam chips were documented and the producers' preference criteria for yam cultivars used to produce chips were identified and prioritized. To further promote the production of yam chips, diversification of insect-resistant yam varieties, conception, and use of health-protective natural insecticides and popularization of modern storage structures were proposed. PMID:23710140

  20. Electromigration in solder joints and solder lines

    NASA Astrophysics Data System (ADS)

    Gan, H.; Choi, W. J.; Xu, G.; Tu, K. N.

    2002-06-01

    Electromigration may affect the reliability of flip-chip solder joints. Eutectic solder is a two-phase alloy, so its electromigration behavior is different from that in aluminum or copper interconnects. In addition, a flipchip solder joint has a built-in currentcrowding configuration to enhance electromigration failure. To better understand electromigration in SnPb and lead-free solder alloys, the authors prepared solder lines in v-grooves etched on Si (001). This article discusses the results of those tests and compares the electromigration failure modes of eutectic SnPb and SnAgCu flip-chip solder joints along with the mean-timeto-failure.

  1. Survival of emerald ash borer in chips

    Treesearch

    Deborah G. McCullough; Therese M. Poland; David L. Cappaert

    2005-01-01

    The ability of emerald ash borer (EAB), Agrilus planipennis Fairmaire, to survive following chipping or grinding of infested ash trees remains a critical question for regulatory officials. In October 2002, we felled eight infested ash trees and sampled sections of the trunk and large branches from each tree to estimate EAB density.

  2. Microarray-based comparison of three amplification methods for nanogram amounts of total RNA

    PubMed Central

    Singh, Ruchira; Maganti, Rajanikanth J.; Jabba, Sairam V.; Wang, Martin; Deng, Glenn; Heath, Joe Don; Kurn, Nurith; Wangemann, Philine

    2007-01-01

    Gene expression profiling using microarrays requires microgram amounts of RNA, which limits its direct application for the study of nanogram RNA samples obtained using microdissection, laser capture microscopy, or needle biopsy. A novel system based on Ribo-SPIA technology (RS, Ovation-Biotin amplification and labeling system) was recently introduced. The utility of the RS system, an optimized prototype system for picogram RNA samples (pRS), and two T7-based systems involving one or two rounds of amplification (OneRA, Standard Protocol, or TwoRA, Small Sample Prototcol, version II) were evaluated in the present study. Mouse kidney (MK) and mouse universal reference (MUR) RNA samples, 0.3 ng to 10 μg, were analyzed using high-density Affymetrix Mouse Genome 430 2.0 GeneChip arrays. Call concordance between replicates, correlations of signal intensity, signal intensity ratios, and minimal fold increase necessary for significance were determined. All systems amplified partially overlapping sets of genes with similar signal intensity correlations. pRS amplified the highest number of genes from 10-ng RNA samples. We detected 24 of 26 genes verified by RT-PCR in samples prepared using pRS. TwoRA yielded somewhat higher call concordances than did RS and pRS (91.8% vs. 89.3% and 88.1%, respectively). Although all target preparation methods were suitable, pRS amplified the highest number of targets and was found to be suitable for amplification of as little as 0.3 ng of total RNA. In addition, RS and pRS were faster and simpler to use than the T7-based methods and resulted in the generation of cDNA, which is more stable than cRNA. PMID:15613496

  3. Fabrication of Quench Condensed Thin Films Using an Integrated MEMS Fab on a Chip

    NASA Astrophysics Data System (ADS)

    Lally, Richard; Reeves, Jeremy; Stark, Thomas; Barrett, Lawrence; Bishop, David

    Atomic calligraphy is a microelectromechanical systems (MEMS)-based dynamic stencil nanolithography technique. Integrating MEMS devices into a bonded stacked array of three die provides a unique platform for conducting quench condensed thin film mesoscopic experiments. The atomic calligraphy Fab on a Chip process incorporates metal film sources, electrostatic comb driven stencil plate, mass sensor, temperature sensor, and target surface into one multi-die assembly. Three separate die are created using the PolyMUMPs process and are flip-chip bonded together. A die containing joule heated sources must be prepared with metal for evaporation prior to assembly. A backside etch of the middle/central die exposes the moveable stencil plate allowing the flux to pass through the stencil from the source die to the target die. The chip assembly is mounted in a cryogenic system at ultra-high vacuum for depositing extremely thin films down to single layers of atoms across targeted electrodes. Experiments such as the effect of thin film alloys or added impurities on their superconductivity can be measured in situ with this process.

  4. Front-End Processing of Cell Lysates for Enhanced Chip-Based Detection

    DTIC Science & Technology

    2006-07-28

    manipulation used in lab-on-a-chip devices. A small unknown sample is first mixed with the PNA surfactants (“PNAA”) to tag the DNA targets, and then the...unknown sample is first mixed with the PNA surfactants (hereafter referred to as “PNA amphiphiles” or “PNAA”) to tag the DNA targets, and then the...prolate ellipsoid, and mixed PNAA/SDS micelles form spherical micelles. On addition of complementary DNA, the PNAA/DNA duplexes do not participate in

  5. Interleukin-6 Detection with a Plasmonic Chip

    NASA Astrophysics Data System (ADS)

    Tawa, Keiko; Sumiya, Masashi; Toma, Mana; Sasakawa, Chisato; Sujino, Takuma; Miyaki, Tatsuki; Nakazawa, Hikaru; Umetsu, Mitsuo

    Interleukin-6, a cytokine relating inflammatory and autoimmune activity, was detected with three fluorescence assays using a plasmonic chip. In their assays, the way of surface modification, sample volume, incubation time and mixing solution, were found to influence the detection sensitivity. When the assay was revised in the point of a rapid and easy process, the detection sensitivity was not compromised compared to assays with sufficient sample volume and assay time. To suit the purpose of immunosensing, the assay conditions should be determined.

  6. A 16-Channel Nonparametric Spike Detection ASIC Based on EC-PC Decomposition.

    PubMed

    Wu, Tong; Xu, Jian; Lian, Yong; Khalili, Azam; Rastegarnia, Amir; Guan, Cuntai; Yang, Zhi

    2016-02-01

    In extracellular neural recording experiments, detecting neural spikes is an important step for reliable information decoding. A successful implementation in integrated circuits can achieve substantial data volume reduction, potentially enabling a wireless operation and closed-loop system. In this paper, we report a 16-channel neural spike detection chip based on a customized spike detection method named as exponential component-polynomial component (EC-PC) algorithm. This algorithm features a reliable prediction of spikes by applying a probability threshold. The chip takes raw data as input and outputs three data streams simultaneously: field potentials, band-pass filtered neural data, and spiking probability maps. The algorithm parameters are on-chip configured automatically based on input data, which avoids manual parameter tuning. The chip has been tested with both in vivo experiments for functional verification and bench-top experiments for quantitative performance assessment. The system has a total power consumption of 1.36 mW and occupies an area of 6.71 mm (2) for 16 channels. When tested on synthesized datasets with spikes and noise segments extracted from in vivo preparations and scaled according to required precisions, the chip outperforms other detectors. A credit card sized prototype board is developed to provide power and data management through a USB port.

  7. 3D nanofabrication inside rapid prototyped microfluidic channels showcased by wet-spinning of single micrometre fibres.

    PubMed

    Lölsberg, Jonas; Linkhorst, John; Cinar, Arne; Jans, Alexander; Kuehne, Alexander J C; Wessling, Matthias

    2018-05-01

    Microfluidics is an established multidisciplinary research domain with widespread applications in the fields of medicine, biotechnology and engineering. Conventional production methods of microfluidic chips have been limited to planar structures, preventing the exploitation of truly three-dimensional architectures for applications such as multi-phase droplet preparation or wet-phase fibre spinning. Here the challenge of nanofabrication inside a microfluidic chip is tackled for the showcase of a spider-inspired spinneret. Multiphoton lithography, an additive manufacturing method, was used to produce free-form microfluidic masters, subsequently replicated by soft lithography. Into the resulting microfluidic device, a three-dimensional spider-inspired spinneret was directly fabricated in-chip via multiphoton lithography. Applying this unprecedented fabrication strategy, the to date smallest printed spinneret nozzle is produced. This spinneret resides tightly sealed, connecting it to the macroscopic world. Its functionality is demonstrated by wet-spinning of single-digit micron fibres through a polyacrylonitrile coagulation process induced by a water sheath layer. The methodology developed here demonstrates fabrication strategies to interface complex architectures into classical microfluidic platforms. Using multiphoton lithography for in-chip fabrication adopts a high spatial resolution technology for improving geometry and thus flow control inside microfluidic chips. The showcased fabrication methodology is generic and will be applicable to multiple challenges in fluid control and beyond.

  8. Rapid and direct detection of attomole adenosine triphosphate (ATP) by MALDI-MS using rutile titania chips.

    PubMed

    Manikandan, Muthu; Hasan, Nazim; Wu, Hui-Fen

    2012-11-07

    We report the rutile titania-based capture of ATP and its application as a MALDI-MS target plate. This chip, when immersed in solutions containing different concentrations of ATP, can capture ATP and lead to its successful detection in MALDI-MS. We have optimized the ideal surface, showing an increased capture efficacy of the 900 °C (rutile) titania surfaces. We demonstrate the use of this chip as a target plate for direct analysis of the attached ATP using MALDI-MS, down to attomolar concentrations. This chip has a promising future for the detection of ATP in environmental samples, which may eventually be used as a pollution indicator in particular environments.

  9. How to Combine ChIP with qPCR.

    PubMed

    Asp, Patrik

    2018-01-01

    Chromatin immunoprecipitation (ChIP) coupled with quantitative PCR (qPCR) has in the last 15 years become a basic mainstream tool in genomic research. Numerous commercially available ChIP kits, qPCR kits, and real-time PCR systems allow for quick and easy analysis of virtually anything chromatin-related as long as there is an available antibody. However, the highly accurate quantitative dimension added by using qPCR to analyze ChIP samples significantly raises the bar in terms of experimental accuracy, appropriate controls, data analysis, and data presentation. This chapter will address these potential pitfalls by providing protocols and procedures that address the difficulties inherent in ChIP-qPCR assays.

  10. A Spiking Strategy for ChIP-chip Data Normalization in S. cerevisiae.

    PubMed

    Jeronimo, Célia; Robert, François

    2017-01-01

    Chromatin immunoprecipitation coupled to DNA microarrays (ChIP-chip) is widely used in the chromatin field, notably to map the position of histone variants or histone modifications along the genome. Often, the position and the occupancy of these epigenetic marks are to be compared between different experiments. It is now increasingly recognized that such cross-sample comparison is better done using externally added exogenous controls for normalization but no such method has been described for ChIP-chip. Here we describe a spiking normalization strategy that makes use of phiX174 phage DNA as a spiked control for normalization of ChIP-chip signals across different experiments.

  11. Low voltage electrophoresis chip with multi-segments synchronized scanning

    NASA Astrophysics Data System (ADS)

    Gu, Wenwen; Wen, Zhiyu; Xu, Yi

    2017-03-01

    For low voltage electrophoresis chip, there is always a problem that the samples are truncated and peaks are broadened, as well as longer time for separation. In this paper, a low voltage electrophoresis separation model was established, and the separation conditions were discussed. A new driving mode was proposed for applying low voltage, which was called multi-segments synchronized scanning. By using this driving mode, the reversed electric field that existed between the multi-segments can enrich samples and shorten the sample zone. The low voltage electrophoresis experiments using multi-segments synchronized scanning were carried out by home-made silicon-PDMS-based chip. The fluorescein isothiocyanate (FITC) labeled lysine and phenylalanine mixed samples with the concentration of 10-4 mol/L were successfully separated under the optimal conditions of 10 mmol/L borax buffer (pH = 10.0), 200 V/cm separation electric field and electrode switch time of 2.5 s. The separation was completed with a resolution of 2.0, and the peak time for lysine and phenylalanine was 4 min and 6 min, respectively.

  12. Simultaneous analysis of seven oligopeptides in microbial fuel cell by micro-fluidic chip with reflux injection mode.

    PubMed

    Wang, Wei; Wang, Zijian; Lin, Xiuli; Wang, ZongWen; Fu, FengFu

    2012-10-15

    In this work, a reflux injection mode for the cross form micro-fluidic chip was studied. This injection mode could flexibly control the length of sample plug from less than one channel width (<83 μm) to tens of channel widths (millimeter-sized) by adjusting the injection time. Namely, the separation resolution or sample detection sensitivity could be selectively improved by changing injection time. Composed of four steps, the reflux injection mode alleviated the electrophoretic sampling bias and prevented sample leakage successfully. On a micro-fluidic chip coupled with laser induced fluorescence (LIF) detector, the injection mode was applied to separate seven oligopeptides, namely GG, GL, RPP, KPV, VKK, WYD and YWS. All analytes were completely separated and detected within 12 min with detection limits of 25-625 nmol/L. At last, the proposed method had been successfully applied to detect oligopeptides consumed by bacillus licheniformis in anode chamber of microbial fuel cell (MFC) to study the effect of oligopeptides on the MFC running. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Effect of Vacuum Frying on Changes in Quality Attributes of Jackfruit (Artocarpus heterophyllus) Bulb Slices

    PubMed Central

    Bawa, A. S.; Raju, P. S.

    2014-01-01

    The effect of frying temperatures and durations on the quality of vacuum fried jackfruit (JF) chips was evaluated. Moisture content and breaking force of JF chips decreased with increase in frying temperature and time during vacuum frying whereas the oil content increased. The frying time for JF chips was found to be 30, 25, and 20 minutes at 80, 90, and 100°C, respectively. JF chips fried at higher temperature resulted in maximum shrinkage (48%). The lightness in terms of hunter L * value decreased significantly (P < 0.05) during frying. Sensory evaluation showed maximum acceptability for JF chips fried at 90°C for 25 min. Frying under vacuum at lower temperatures was found to retain bioactive compounds such as total phenolics, total flavonoids, and total carotenoids in JF chips. Almost 90% of carotenoids were lost from the samples after 30 min of frying at 100°C. PMID:26904648

  14. Digital PCR on a SlipChip.

    PubMed

    Shen, Feng; Du, Wenbin; Kreutz, Jason E; Fok, Alice; Ismagilov, Rustem F

    2010-10-21

    This paper describes a SlipChip to perform digital PCR in a very simple and inexpensive format. The fluidic path for introducing the sample combined with the PCR mixture was formed using elongated wells in the two plates of the SlipChip designed to overlap during sample loading. This fluidic path was broken up by simple slipping of the two plates that removed the overlap among wells and brought each well in contact with a reservoir preloaded with oil to generate 1280 reaction compartments (2.6 nL each) simultaneously. After thermal cycling, end-point fluorescence intensity was used to detect the presence of nucleic acid. Digital PCR on the SlipChip was tested quantitatively by using Staphylococcus aureus genomic DNA. As the concentration of the template DNA in the reaction mixture was diluted, the fraction of positive wells decreased as expected from the statistical analysis. No cross-contamination was observed during the experiments. At the extremes of the dynamic range of digital PCR the standard confidence interval determined using a normal approximation of the binomial distribution is not satisfactory. Therefore, statistical analysis based on the score method was used to establish these confidence intervals. The SlipChip provides a simple strategy to count nucleic acids by using PCR. It may find applications in research applications such as single cell analysis, prenatal diagnostics, and point-of-care diagnostics. SlipChip would become valuable for diagnostics, including applications in resource-limited areas after integration with isothermal nucleic acid amplification technologies and visual readout.

  15. A fully integrated distance readout ELISA-Chip for point-of-care testing with sample-in-answer-out capability.

    PubMed

    Liu, Dan; Li, Xingrui; Zhou, Junkai; Liu, Shibo; Tian, Tian; Song, Yanling; Zhu, Zhi; Zhou, Leiji; Ji, Tianhai; Yang, Chaoyong

    2017-10-15

    Enzyme-linked immunosorbent assay (ELISA) is a popular laboratory technique for detection of disease-specific protein biomarkers with high specificity and sensitivity. However, ELISA requires labor-intensive and time-consuming procedures with skilled operators and spectroscopic instrumentation. Simplification of the procedures and miniaturization of the devices are crucial for ELISA-based point-of-care (POC) testing in resource-limited settings. Here, we present a fully integrated, instrument-free, low-cost and portable POC platform which integrates the process of ELISA and the distance readout into a single microfluidic chip. Based on manipulation using a permanent magnet, the process is initiated by moving magnetic beads with capture antibody through different aqueous phases containing ELISA reagents to form bead/antibody/antigen/antibody sandwich structure, and finally converts the molecular recognition signal into a highly sensitive distance readout for visual quantitative bioanalysis. Without additional equipment and complicated operations, our integrated ELISA-Chip with distance readout allows ultrasensitive quantitation of disease biomarkers within 2h. The ELISA-Chip method also showed high specificity, good precision and great accuracy. Furthermore, the ELISA-Chip system is highly applicable as a sandwich-based platform for the detection of a variety of protein biomarkers. With the advantages of visual analysis, easy operation, high sensitivity, and low cost, the integrated sample-in-answer-out ELISA-Chip with distance readout shows great potential for quantitative POCT in resource-limited settings. Copyright © 2017. Published by Elsevier B.V.

  16. Ultra-Sensitive Lab-on-a-Chip Detection of Sudan I in Food using Plasmonics-Enhanced Diatomaceous Thin Film.

    PubMed

    Kong, Xianming; Squire, Kenny; Chong, Xinyuan; Wang, Alan X

    2017-09-01

    Sudan I is a carcinogenic compound containing an azo group that has been illegally utilized as an adulterant in food products to impart a bright red color to foods. In this paper, we develop a facile lab-on-a-chip device for instant, ultra-sensitive detection of Sudan I from real food samples using plasmonics-enhanced diatomaceous thin film, which can simultaneously perform on-chip separation using thin layer chromatography (TLC) and highly specific sensing using surface-enhanced Raman scattering (SERS) spectroscopy. Diatomite is a kind of nature-created photonic crystal biosilica with periodic pores and was used both as the stationary phase of the TLC plate and photonic crystals to enhance the SERS sensitivity. The on-chip chromatography capability of the TLC plate was verified by isolating Sudan I in a mixture solution containing Rhodamine 6G, while SERS sensing was achieved by spraying gold colloidal nanoparticles into the sensing spot. Such plasmonics-enhanced diatomaceous film can effectively detect Sudan I with more than 10 times improvement of the Raman signal intensity than commercial silica gel TLC plates. We applied this lab-on-a-chip device for real food samples and successfully detected Sudan I in chili sauce and chili oil down to 1 ppm, or 0.5 ng/spot. This on-chip TLC-SERS biosensor based on diatomite biosilica can function as a cost-effective, ultra-sensitive, and reliable technology for screening Sudan I and many other illicit ingredients to enhance food safety.

  17. Effect of structural evolution of ZnO/HfO2 nanocrystals on Eu2+/Eu3+ emission in glass-ceramic waveguides for photonic applications

    NASA Astrophysics Data System (ADS)

    Ghosh, Subhabrata; N, Shivakiran Bhaktha B.

    2018-06-01

    Eu-doped 70SiO2–23HfO2–7ZnO (mol%) glass-ceramic waveguides have been fabricated by sol-gel method as a function of heat-treatment temperatures for on-chip blue-light emitting source applications. Structural evolution of spherical ZnO and spherical as well as rod-like HfO2 nanocrystalline structures have been observed with heat-treatments at different temperatures. Initially, in the as-prepared samples at 900 ◦C, both, Eu2+ as well as Eu3+ ions are found to be present in the ternary matrix. With controlled heat-treatments of up to 1000 ◦C for 2 h, local environment of Eu-ions become more crystalline in nature and the reduction of Eu3+ to Eu2+ takes place in such ZnO/HfO2 crystalline environments. In these ternary glass-ceramic waveguides, heat-treated at higher temperatures, the blue-light emission characteristic, which is the signature of 4f 65d \\to 4f 7 energy level transition of Eu2+ ions is found to be greatly enhanced. The as-prepared glass-ceramic waveguides exhibit a propagation loss of 0.4 ± 0.2 dB cm‑1 at 632.8 nm. Though the propagation losses increase with the growth of nanocrystals, the added functionalities achieved in the optimally heat-treated Eu-doped 70SiO2–23HfO2–7ZnO (mol%) waveguides, make them a viable functional optical material for the fabrication of on-chip blue-light emitting sources for integrated optic applications.

  18. Effect of structural evolution of ZnO/HfO2 nanocrystals on Eu2+/Eu3+ emission in glass-ceramic waveguides for photonic applications.

    PubMed

    Ghosh, Subhabrata; Bhaktha B N, Shivakiran

    2018-06-01

    Eu-doped 70SiO 2 -23HfO 2 -7ZnO (mol%) glass-ceramic waveguides have been fabricated by sol-gel method as a function of heat-treatment temperatures for on-chip blue-light emitting source applications. Structural evolution of spherical ZnO and spherical as well as rod-like HfO 2 nanocrystalline structures have been observed with heat-treatments at different temperatures. Initially, in the as-prepared samples at 900 ◦ C, both, Eu 2+ as well as Eu 3+ ions are found to be present in the ternary matrix. With controlled heat-treatments of up to 1000 ◦ C for 2 h, local environment of Eu-ions become more crystalline in nature and the reduction of Eu 3+ to Eu 2+ takes place in such ZnO/HfO 2 crystalline environments. In these ternary glass-ceramic waveguides, heat-treated at higher temperatures, the blue-light emission characteristic, which is the signature of 4f 6 5d [Formula: see text] 4f 7 energy level transition of Eu 2+ ions is found to be greatly enhanced. The as-prepared glass-ceramic waveguides exhibit a propagation loss of 0.4 ± 0.2 dB cm -1 at 632.8 nm. Though the propagation losses increase with the growth of nanocrystals, the added functionalities achieved in the optimally heat-treated Eu-doped 70SiO 2 -23HfO 2 -7ZnO (mol%) waveguides, make them a viable functional optical material for the fabrication of on-chip blue-light emitting sources for integrated optic applications.

  19. A microchip-based flow injection-amperometry system with mercaptopropionic acid modified electroless gold microelectrode for the selective determination of dopamine.

    PubMed

    Wang, Yi; Luo, Jie; Chen, Hengwu; He, Qiaohong; Gan, Nin; Li, Tianhua

    2008-09-12

    A novel chip-based flow injection analysis (FIA) system has been developed for automatic, rapid and selective determination of dopamine (DA) in the presence of ascorbic acid (AA). The system is composed of a polycarbonate (PC) microfluidic chip with an electrochemical detector (ED), a gravity pump, and an automatic sample loading and injection unit. The selectivity of the ED was improved by modification of the gold working microelectrode, which was fabricated on the PC chip by UV-directed electroless gold plating, with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA). Postplating treatment methods for cleaning the surface of electroless gold microelectrodes were investigated to ensure the formation of high quality SAMs. The effects of detection potential, flow rate, and sampling volume on the performance of the chip-based FIA system were studied. Under optimum conditions, a detection limit of 74 nmol L(-1) for DA was achieved at the sample throughput rate of 180 h(-1). A RSD of 0.9% for peak heights was observed for 19 runs of a 100 micromol L(-1) DA solution. Interference-free determination of DA could be conducted if the concentration ratio of AA-DA was no more than 10.

  20. A versatile electrophoresis-based self-test platform.

    PubMed

    Staal, Steven; Ungerer, Mathijn; Floris, Arjan; Ten Brinke, Hans-Willem; Helmhout, Roy; Tellegen, Marian; Janssen, Kjeld; Karstens, Erik; van Arragon, Charlotte; Lenk, Stefan; Staijen, Erik; Bartholomew, Jody; Krabbe, Hans; Movig, Kris; Dubský, Pavel; van den Berg, Albert; Eijkel, Jan

    2015-03-01

    This paper reports on recent research creating a family of electrophoresis-based point of care devices for the determination of a wide range of ionic analytes in various sample matrices. These devices are based on a first version for the point-of-care measurement of Li(+), reported in 2010 by Floris et al. (Lab Chip 2010, 10, 1799-1806). With respect to this device, significant improvements in accuracy, precision, detection limit, and reliability have been obtained especially by the use of multiple injections of one sample on a single chip and integrated data analysis. Internal and external validation by clinical laboratories for the determination of analytes in real patients by a self-test is reported. For Li(+) in blood better precision than the standard clinical determination for Li(+) was achieved. For Na(+) in human urine the method was found to be within the clinical acceptability limits. In a veterinary application, Ca(2+) and Mg(2+) were determined in bovine blood by means of the same chip, but using a different platform. Finally, promising preliminary results are reported with the Medimate platform for the determination of creatinine in whole blood and quantification of both cations and anions through replicate measurements on the same sample with the same chip. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Plug-and-play, infrared, laser-mediated PCR in a microfluidic chip.

    PubMed

    Pak, Nikita; Saunders, D Curtis; Phaneuf, Christopher R; Forest, Craig R

    2012-04-01

    Microfluidic polymerase chain reaction (PCR) systems have set milestones for small volume (100 nL-5 μL), amplification speed (100-400 s), and on-chip integration of upstream and downstream sample handling including purification and electrophoretic separation functionality. In practice, the microfluidic chips in these systems require either insertion of thermocouples or calibration prior to every amplification. These factors can offset the speed advantages of microfluidic PCR and have likely hindered commercialization. We present an infrared, laser-mediated, PCR system that features a single calibration, accurate and repeatable precision alignment, and systematic thermal modeling and management for reproducible, open-loop control of PCR in 1 μL chambers of a polymer microfluidic chip. Total cycle time is less than 12 min: 1 min to fill and seal, 10 min to amplify, and 1 min to recover the sample. We describe the design, basis for its operation, and the precision engineering in the system and microfluidic chip. From a single calibration, we demonstrate PCR amplification of a 500 bp amplicon from λ-phage DNA in multiple consecutive trials on the same instrument as well as multiple identical instruments. This simple, relatively low-cost plug-and-play design is thus accessible to persons who may not be skilled in assembly and engineering.

  2. Best Practices and Joint Calling of the HumanExome BeadChip: The CHARGE Consortium

    PubMed Central

    Grove, Megan L.; Yu, Bing; Cochran, Barbara J.; Haritunians, Talin; Bis, Joshua C.; Taylor, Kent D.; Hansen, Mark; Borecki, Ingrid B.; Cupples, L. Adrienne; Fornage, Myriam; Gudnason, Vilmundur; Harris, Tamara B.; Kathiresan, Sekar; Kraaij, Robert; Launer, Lenore J.; Levy, Daniel; Liu, Yongmei; Mosley, Thomas; Peloso, Gina M.; Psaty, Bruce M.; Rich, Stephen S.; Rivadeneira, Fernando; Siscovick, David S.; Smith, Albert V.; Uitterlinden, Andre; van Duijn, Cornelia M.; Wilson, James G.; O’Donnell, Christopher J.; Rotter, Jerome I.; Boerwinkle, Eric

    2013-01-01

    Genotyping arrays are a cost effective approach when typing previously-identified genetic polymorphisms in large numbers of samples. One limitation of genotyping arrays with rare variants (e.g., minor allele frequency [MAF] <0.01) is the difficulty that automated clustering algorithms have to accurately detect and assign genotype calls. Combining intensity data from large numbers of samples may increase the ability to accurately call the genotypes of rare variants. Approximately 62,000 ethnically diverse samples from eleven Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) Consortium cohorts were genotyped with the Illumina HumanExome BeadChip across seven genotyping centers. The raw data files for the samples were assembled into a single project for joint calling. To assess the quality of the joint calling, concordance of genotypes in a subset of individuals having both exome chip and exome sequence data was analyzed. After exclusion of low performing SNPs on the exome chip and non-overlap of SNPs derived from sequence data, genotypes of 185,119 variants (11,356 were monomorphic) were compared in 530 individuals that had whole exome sequence data. A total of 98,113,070 pairs of genotypes were tested and 99.77% were concordant, 0.14% had missing data, and 0.09% were discordant. We report that joint calling allows the ability to accurately genotype rare variation using array technology when large sample sizes are available and best practices are followed. The cluster file from this experiment is available at www.chargeconsortium.com/main/exomechip. PMID:23874508

  3. Dietary acrylamide exposure of the French population: results of the second French Total Diet Study.

    PubMed

    Sirot, Véronique; Hommet, Frédéric; Tard, Alexandra; Leblanc, Jean-Charles

    2012-03-01

    Acrylamide is a heat-induced carcinogen compound that is found in some foods consequently to cooking or other thermal processes. In the second French Total Diet Study (TDS), acrylamide was analysed in 192 food samples collected in mainland France to be representative of the population diet and prepared "as consumed". Highest mean concentrations were found in potato chips/crisps (954 μg/kg), French fries and other fried potatoes (724 μg/kg), and salted biscuits other than potato chips (697 μg/kg). Exposure of general adult and child populations was assessed by combining analytical results with national consumption data. Mean acrylamide exposure was assessed to be 0.43±0.33 μg/kg of body weight (bw) per day for adults and 0.69±0.58 μg/kg bw/day for children. Although the exposure assessed is lower than in previous evaluations, the calculated margins of exposure, based on benchmark dose limits defined for carcinogenic effects, remain very low especially for young children (below 100 at the 95th percentile of exposure), indicating a health concern. It is therefore advisable to continue efforts in order to reduce dietary exposure to acrylamide. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Clean galena, contaminated lead, and soft errors in memory chips

    NASA Astrophysics Data System (ADS)

    Lykken, G. I.; Hustoft, J.; Ziegler, B.; Momcilovic, B.

    2000-10-01

    Lead (Pb) disks were exposed to a radon (Rn)-rich atmosphere and surface alpha particle emissions were detected over time. Cumulative 210Po alpha emission increased nearly linearly with time. Conversely, cumulative emission for each of 218Po and 214Po was constant after one and two hours, respectively. Processing of radiation-free Pb ore (galena) in inert atmospheres was compared with processing in ambient air. Galena processed within a flux heated in a graphite crucible while exposed to an inert atmosphere, resulted in lead contaminated with 210Po (Trial 1). A glove box was next used to prepare a baseline radiation-free flux sample in an alumina crucible that was heated in an oven with an inert atmosphere (Trials 2 and 3). Ambient air was thereafter introduced, in place of the inert atmosphere, to the radiation-free flux mixture during processing (Trial 4). Ambient air introduced Rn and its progeny (RAD) into the flux during processing so that the processed Pb contained Po isotopes. A typical coke used in lead smelting also emitted numerous alpha particles. We postulate that alpha particles from tin/lead solder bumps, a cause of computer chip memory soft errors, may originate from Rn and RAD in the ambient air and/or coke used as a reducing agent in the standard galena smelting procedure.

  5. Effect of Biodegradable Scleral Plugs Containing Curcumin on Proliferative Vitreoretinopathy.

    PubMed

    Zhang, Jun; Zhou, Nalei; Zhang, Bin; Ma, Jingxue

    2018-01-01

    This study aimed to explore the inhibitory effect of biodegradable scleral plugs containing curcumin on rabbits with proliferative vitreoretinopathy (PVR). The biodegradable scleral plugs containing curcumin were prepared by dissolving PLGA [poly(lactide-co-glycolide)] and curcumin. In total, 30 rabbits were divided into 2 groups: the model group received a vitreous injection of self-blood, and the treatment group received a vitreous injection of self-blood plus biodegradable scleral implants containing 1.5 mg of curcumin. On days 1, 3, 7, 14, 21, and 28 after the operation, clinical observations and PVR classifications were performed. Then, after vitreous samples were collected, different cytokines were detected using antibody chip technology. The scleral plug was 5 mm in length and 1 mm in diameter. Clinical observation showed marked inflammation in the model group. The development grade of PVR in the treatment group was lower than that in the model group (p < 0.05). The outcome of antibody chip technology showed that the expression levels of IL-1α, IL-1β, IL-8, leptin, MMP-9, NCAM, and TNF-α in the treatment group at different time points were significantly lower than those in the model group (p < 0.05). Curcumin might have great potential as a therapeutic agent for PVR by inhibiting various inflammatory factors. © 2017 S. Karger AG, Basel.

  6. Microfluidic desalination techniques and their potential applications.

    PubMed

    Roelofs, S H; van den Berg, A; Odijk, M

    2015-09-07

    In this review we discuss recent developments in the emerging research field of miniaturized desalination. Traditionally desalination is performed to convert salt water into potable water and research is focused on improving performance of large-scale desalination plants. Microfluidic desalination offers several new opportunities in comparison to macro-scale desalination, such as providing a platform to increase fundamental knowledge of ion transport on the nano- and microfluidic scale and new microfluidic sample preparation methods. This approach has also lead to the development of new desalination techniques, based on micro/nanofluidic ion-transport phenomena, which are potential candidates for up-scaling to (portable) drinking water devices. This review assesses microfluidic desalination techniques on their applications and is meant to contribute to further implementation of microfluidic desalination techniques in the lab-on-chip community.

  7. Making Shock Waves in Microfluidics: The Physics and Applications of Isotachophoresis

    NASA Astrophysics Data System (ADS)

    Santiago, Juan

    2007-11-01

    Microfluidics lies at the interfaces between engineering, chemistry, and biology, and aims to develop chemical laboratories on a chip. An important technique is on-chip capillary electrophoresis which has been applied to a wide range of chemical and biochemical assay applications over the last decade. Perhaps the best way of improving the sensitivity of on-chip electrophoresis is to integrate an online sample preconcentration method. At Stanford, we are developing methods to concentrate ions into small volumes using a method called isotachophoresis (ITP). In ITP, sample ions are injected between the high mobility co-ions of a leading electrolyte (LE) and the low mobility co-ions of a trailing electrolyte (TE). Upon application of an electric field, the disparate ion mobilities of the LE and TE cause sample species to segregate and focus into a series of narrow self-sharpening zones which migrate at equal velocity (hence ``isotacho''). ITP-type processes have been studied and used for more than 60 years, and yet there remain significant challenges in the robust modeling of these transport processes and the creation of widely applicable assays. We use ITP to create sample ion concentration ``shock waves'' in microchannels. These concentration waves can be integrated with on-chip electrophoresis for high sensitivity assays, and novel modes of operation. The talk will summarize the basic physics of ITP, experimental studies of ITP, models of ITP, and the development of novel ITP-assays with unprecedented sensitivity and new functionality. For example, using leading-to-sample ion concentration ratios of 10^15 and local electric fields of ˜4 kV/cm, we can achieve order one micron wide ITP zones. We can achieve million fold preconcentration in 120 s and can detect 100 attomolar sample concentrations (to our knowledge the highest demonstrated sensitivity for an electrophoresis-related assay). We have also developed a method that uses ITP to separate, indirectly detect, and identify the electrophoretic mobilities of unlabeled (non-fluorescent) analytes using surrogate fluorescent molecules. Our goal is the development of novel on-chip ITP assays which expand the design space of microfluidic devices.

  8. On-chip supercapacitors with ultrahigh volumetric performance based on electrochemically co-deposited CuO/polypyrrole nanosheet arrays

    NASA Astrophysics Data System (ADS)

    Qian, Tao; Zhou, Jinqiu; Xu, Na; Yang, Tingzhou; Shen, Xiaowei; Liu, Xuejun; Wu, Shishan; Yan, Chenglin

    2015-10-01

    We introduce a new method for fabricating unique on-chip supercapacitors based on CuO/polypyrrole core/shell nanosheet arrays by means of direct electrochemical co-deposition on interdigital-like electrodes. The prepared all-solid-state device demonstrates exceptionally high specific capacitance of 1275.5 F cm-3 (˜40 times larger than that of CuO-only supercapacitors) and high-energy-density of 28.35 mWh cm-3, which are both significantly greater than other solid-state supercapacitors. More importantly, the device maintains approximately 100% capacity retention at 2.5 A cm-3 after 3000 cycles. The in situ co-deposition of CuO/polypyrrole nanosheets on interdigital substrate enables effective charge transport, electrode fabrication integrity, and device integration. Because of their high energy, power density, and stable cycling stability, these newly developed on-chip supercapacitors permit fast, reliable applications in portable and miniaturized electronic devices.

  9. On-chip supercapacitors with ultrahigh volumetric performance based on electrochemically co-deposited CuO/polypyrrole nanosheet arrays.

    PubMed

    Qian, Tao; Zhou, Jinqiu; Xu, Na; Yang, Tingzhou; Shen, Xiaowei; Liu, Xuejun; Wu, Shishan; Yan, Chenglin

    2015-10-23

    We introduce a new method for fabricating unique on-chip supercapacitors based on CuO/polypyrrole core/shell nanosheet arrays by means of direct electrochemical co-deposition on interdigital-like electrodes. The prepared all-solid-state device demonstrates exceptionally high specific capacitance of 1275.5 F cm(-3) (∼40 times larger than that of CuO-only supercapacitors) and high-energy-density of 28.35 mWh cm(-3), which are both significantly greater than other solid-state supercapacitors. More importantly, the device maintains approximately 100% capacity retention at 2.5 A cm(-3) after 3000 cycles. The in situ co-deposition of CuO/polypyrrole nanosheets on interdigital substrate enables effective charge transport, electrode fabrication integrity, and device integration. Because of their high energy, power density, and stable cycling stability, these newly developed on-chip supercapacitors permit fast, reliable applications in portable and miniaturized electronic devices.

  10. On-chip low loss heralded source of pure single photons.

    PubMed

    Spring, Justin B; Salter, Patrick S; Metcalf, Benjamin J; Humphreys, Peter C; Moore, Merritt; Thomas-Peter, Nicholas; Barbieri, Marco; Jin, Xian-Min; Langford, Nathan K; Kolthammer, W Steven; Booth, Martin J; Walmsley, Ian A

    2013-06-03

    A key obstacle to the experimental realization of many photonic quantum-enhanced technologies is the lack of low-loss sources of single photons in pure quantum states. We demonstrate a promising solution: generation of heralded single photons in a silica photonic chip by spontaneous four-wave mixing. A heralding efficiency of 40%, corresponding to a preparation efficiency of 80% accounting for detector performance, is achieved due to efficient coupling of the low-loss source to optical fibers. A single photon purity of 0.86 is measured from the source number statistics without narrow spectral filtering, and confirmed by direct measurement of the joint spectral intensity. We calculate that similar high-heralded-purity output can be obtained from visible to telecom spectral regions using this approach. On-chip silica sources can have immediate application in a wide range of single-photon quantum optics applications which employ silica photonics.

  11. DNA microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines.

    PubMed

    Yamagishi, J; Isobe, R; Takebuchi, T; Bando, H

    2003-03-01

    We describe, for the first time, the generation of a viral DNA chip for simultaneous expression measurements of nearly all known open reading frames (ORFs) in the best-studied members of the family Baculoviridae, Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV). In this study, a viral DNA chip (Ac-BmNPV chip) was fabricated and used to characterize the viral gene expression profile for AcMNPV in different cell types. The viral chip is composed of microarrays of viral DNA prepared by robotic deposition of PCR-amplified viral DNA fragments on glass for ORFs in the NPV genome. Viral gene expression was monitored by hybridization to the DNA fragment microarrays with fluorescently labeled cDNAs prepared from infected Spodoptera frugiperda, Sf9 cells and Trichoplusia ni, TnHigh-Five cells, the latter a major producer of baculovirus and recombinant proteins. A comparison of expression profiles of known ORFs in AcMNPV elucidated six genes (ORF150, p10, pk2, and three late gene expression factor genes lef-3, p35 and lef- 6) the expression of each of which was regulated differently in the two cell lines. Most of these genes are known to be closely involved in the viral life cycle such as in DNA replication, late gene expression and the release of polyhedra from infected cells. These results imply that the differential expression of these viral genes accounts for the differences in viral replication between these two cell lines. Thus, these fabricated microarrays of NPV DNA which allow a rapid analysis of gene expression at the viral genome level should greatly speed the functional analysis of large genomes of NPV.

  12. Children with Special Health Care Needs in CHIP: Access, Use, and Child and Family Outcomes.

    PubMed

    Zickafoose, Joseph S; Smith, Kimberly V; Dye, Claire

    2015-01-01

    To assess how the Children's Health Insurance Program (CHIP) affects outcomes for children with special health care needs (CSHCN). We used data from a survey of parents of recent and established CHIP enrollees conducted from January 2012 through March 2013 as part of a congressionally mandated evaluation of CHIP. We identified CSHCN in the sample using the Child and Adolescent Health Measurement Initiative's CSHCN screener. We compared the health care experiences of established CHIP enrollees to the pre-enrollment experiences of previously uninsured and privately insured recent CHIP enrollees, controlling for observable characteristics. Parents of 4142 recent enrollees and 5518 established enrollees responded to the survey (response rates, 46% recent enrollees and 51% established enrollees). In the 10 survey states, about one-fourth of CHIP enrollees had a special health care need. Compared to being uninsured, parents of CSHCN who were established CHIP enrollees reported greater access to and use of medical and dental care, less difficulty meeting their child's health care needs, fewer unmet needs, and better dental health status for their child. Compared to having private insurance, parents of CSHCN who were established CHIP enrollees reported similar levels of access to and use of medical and dental care and unmet needs, and less difficulty meeting their child's health care needs. CHIP has significant benefits for eligible CSHCN and their families compared to being uninsured and appears to have some benefits compared to private insurance. Copyright © 2015 Academic Pediatric Association. All rights reserved.

  13. Assessing the Power of Exome Chips.

    PubMed

    Page, Christian Magnus; Baranzini, Sergio E; Mevik, Bjørn-Helge; Bos, Steffan Daniel; Harbo, Hanne F; Andreassen, Bettina Kulle

    2015-01-01

    Genotyping chips for rare and low-frequent variants have recently gained popularity with the introduction of exome chips, but the utility of these chips remains unclear. These chips were designed using exome sequencing data from mainly American-European individuals, enriched for a narrow set of common diseases. In addition, it is well-known that the statistical power of detecting associations with rare and low-frequent variants is much lower compared to studies exclusively involving common variants. We developed a simulation program adaptable to any exome chip design to empirically evaluate the power of the exome chips. We implemented the main properties of the Illumina HumanExome BeadChip array. The simulated data sets were used to assess the power of exome chip based studies for varying effect sizes and causal variant scenarios. We applied two widely-used statistical approaches for rare and low-frequency variants, which collapse the variants into genetic regions or genes. Under optimal conditions, we found that a sample size between 20,000 to 30,000 individuals were needed in order to detect modest effect sizes (0.5% < PAR > 1%) with 80% power. For small effect sizes (PAR <0.5%), 60,000-100,000 individuals were needed in the presence of non-causal variants. In conclusion, we found that at least tens of thousands of individuals are necessary to detect modest effects under optimal conditions. In addition, when using rare variant chips on cohorts or diseases they were not originally designed for, the identification of associated variants or genes will be even more challenging.

  14. Cancer stem-like cell related protein CD166 degrades through E3 ubiquitin ligase CHIP in head and neck cancer.

    PubMed

    Xiao, Meng; Yan, Ming; Zhang, Jianjun; Xu, Qin; Qi, Shengcai; Wang, Xu; Chen, Wantao

    2017-04-01

    Our previous studies have identified that CD166 works as a cancer stem-like cell (CSC) marker in epithelial cancers with a large repertoire of cellular functions. However, the post-translational regulatory mechanisms underlying CD166 turnover remain elusive. Several independent studies have reported that E3 ubiquitin ligase CHIP revealed significant biological effects through ubiquitin proteasome pathway on some kinds of malignant tumors. With analyzing the effects of CHIP expressions on stem-like cell populations, we found that CHIP represses CSC characteristics mainly targeting the CSC related protein CD166 in head and neck cancer (HNC). To investigate the role and relationship between CD166 and CHIP, HNC tissues and cell lines were used in this study. A significant negative correlation was observed between the expression levels of CHIP and CD166 in HNC patient samples. We also found that CHIP directly regulates the stability of CD166 protein through the ubiquitin proteasome system, which was also identified participating in the regulation of CSC behaviors in HNCs. Our findings demonstrate that CHIP-CD166-proteasome axis participates in regulating CSC properties in HNCs, suggesting that the regulation of CD166 by CHIP could provide new options for diagnosing and treating in the patients with HNCs. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  15. Application of allflex conservation buffer in illumina genotyping.

    PubMed

    de Groot, M; Ras, T; van Haeringen, W A

    2016-12-01

    This experiment was designed to study if liquid conservation buffer used in the novel Tissue Sampling Technology (TST) from Allflex can be used for Illumina BeadChip genotyping. Ear punches were collected from 6 bovine samples, using both the Tissue Sampling Unit (TSU) as well as the Total Tagger Universal (TTU) collection system. The stability of the liquid conservation buffer was tested by genotyping samples on Illumina BeadChips, incubated at 0, 3, 15, 24, 48, 72, 168, 336, 720 h after sample collection. Additionally, a replenishment study was designed to test how often the liquid conservation buffer could be completely replenished before a significant call rate drop could be observed. Results from the stability study showed an average call rate of 0.993 for samples collected with the TSU system and 0.953 for samples collected with the TTU system, both exceeding the inclusion threshold call rate of 0.85. As an additional control, the identity of the individual animals was confirmed using the International Society of Animal Genetics (ISAG) recommended SNP panel. The replenishment study revealed a slight drop in the sample call rate after replenishing the conservation buffer for the fourth time for the TSU as well as the TTU samples. In routine analysis, this application allows for multiple experiments to be performed on the liquid conservation buffer, while maintaining the tissue samples for future use. The data collected in this study shows that the liquid conservation buffer used in the TST system can be used for Illumina BeadChip genotyping applications.

  16. VLSI chip-set for data compression using the Rice algorithm

    NASA Technical Reports Server (NTRS)

    Venbrux, J.; Liu, N.

    1990-01-01

    A full custom VLSI implementation of a data compression encoder and decoder which implements the lossless Rice data compression algorithm is discussed in this paper. The encoder and decoder reside on single chips. The data rates are to be 5 and 10 Mega-samples-per-second for the decoder and encoder respectively.

  17. Biofunctionalized self-propelled micromotors as an alternative on-chip concentrating system.

    PubMed

    Restrepo-Pérez, Laura; Soler, Lluís; Martínez-Cisneros, Cynthia; Sánchez, Samuel; Schmidt, Oliver G

    2014-08-21

    Sample pre-concentration is crucial to achieve high sensitivity and low detection limits in lab-on-a-chip devices. Here, we present a system in which self-propelled catalytic micromotors are biofunctionalized and trapped acting as an alternative concentrating mechanism. This system requires no external energy source, which facilitates integration and miniaturization.

  18. Oligonucleotide Microarray for 16S rRNA Gene-Based Detection of All Recognized Lineages of Sulfate-Reducing Prokaryotes in the Environment

    PubMed Central

    Loy, Alexander; Lehner, Angelika; Lee, Natuschka; Adamczyk, Justyna; Meier, Harald; Ernst, Jens; Schleifer, Karl-Heinz; Wagner, Michael

    2002-01-01

    For cultivation-independent detection of sulfate-reducing prokaryotes (SRPs) an oligonucleotide microarray consisting of 132 16S rRNA gene-targeted oligonucleotide probes (18-mers) having hierarchical and parallel (identical) specificity for the detection of all known lineages of sulfate-reducing prokaryotes (SRP-PhyloChip) was designed and subsequently evaluated with 41 suitable pure cultures of SRPs. The applicability of SRP-PhyloChip for diversity screening of SRPs in environmental and clinical samples was tested by using samples from periodontal tooth pockets and from the chemocline of a hypersaline cyanobacterial mat from Solar Lake (Sinai, Egypt). Consistent with previous studies, SRP-PhyloChip indicated the occurrence of Desulfomicrobium spp. in the tooth pockets and the presence of Desulfonema- and Desulfomonile-like SRPs (together with other SRPs) in the chemocline of the mat. The SRP-PhyloChip results were confirmed by several DNA microarray-independent techniques, including specific PCR amplification, cloning, and sequencing of SRP 16S rRNA genes and the genes encoding the dissimilatory (bi)sulfite reductase (dsrAB). PMID:12324358

  19. Microfluidic chip for peptide analysis with an integrated HPLC column, sample enrichment column, and nanoelectrospray tip.

    PubMed

    Yin, Hongfeng; Killeen, Kevin; Brennen, Reid; Sobek, Dan; Werlich, Mark; van de Goor, Tom

    2005-01-15

    Current nano-LC/MS systems require the use of an enrichment column, a separation column, a nanospray tip, and the fittings needed to connect these parts together. In this paper, we present a microfabricated approach to nano-LC, which integrates these components on a single LC chip, eliminating the need for conventional LC connections. The chip was fabricated by laminating polyimide films with laser-ablated channels, ports, and frit structures. The enrichment and separation columns were packed using conventional reversed-phase chromatography particles. A face-seal rotary valve provided a means for switching between sample loading and separation configurations with minimum dead and delay volumes while allowing high-pressure operation. The LC chip and valve assembly were mounted within a custom electrospray source on an ion-trap mass spectrometer. The overall system performance was demonstrated through reversed-phase gradient separations of tryptic protein digests at flow rates between 100 and 400 nL/min. Microfluidic integration of the nano-LC components enabled separations with subfemtomole detection sensitivity, minimal carryover, and robust and stable electrospray throughout the LC solvent gradient.

  20. Smartphone-enabled optofluidic exosome diagnostic for concussion recovery

    PubMed Central

    Ko, Jina; Hemphill, Matthew A.; Gabrieli, David; Wu, Leon; Yelleswarapu, Venkata; Lawrence, Gladys; Pennycooke, Wesley; Singh, Anup; Meaney, Dave F.; Issadore, David

    2016-01-01

    A major impediment to improving the treatment of concussion is our current inability to identify patients that will experience persistent problems after the injury. Recently, brain-derived exosomes, which cross the blood-brain barrier and circulate following injury, have shown great potential as a noninvasive biomarker of brain recovery. However, clinical use of exosomes has been constrained by their small size (30–100 nm) and the extensive sample preparation (>24 hr) needed for traditional exosome measurements. To address these challenges, we developed a smartphone-enabled optofluidic platform to measure brain-derived exosomes. Sample-to-answer on our chip is 1 hour, 10x faster than conventional techniques. The key innovation is an optofluidic device that can detect enzyme amplified exosome biomarkers, and is read out using a smartphone camera. Using this approach, we detected and profiled GluR2+ exosomes in the post-injury state using both in vitro and murine models of concussion. PMID:27498963

  1. Acrylamide in Caribbean foods - residual levels and their relation to reducing sugar and asparagine content.

    PubMed

    Bent, Grace-Anne; Maragh, Paul; Dasgupta, Tara

    2012-07-15

    The acrylamide levels in commercial and homemade Caribbean foods were determined by pre-derivatisation of acrylamide to 2-bromopropenamide and analysed by gas chromatography with mass spectrometric (GC/MS) detection. Over 100 Caribbean food samples were analysed for the presence of acrylamide. These samples include: biscuits, breakfast cereals, banana chips and home-prepared foods: breadfruit; Artocarpus altilis, banana fritters, and dumplings. The limit of detection (LOD) for the GC/MS method was found to be dependent on the type of column used for the GC/MS analysis. The DB-1701 and the DB-VRX columns gave LODs of 20 and 4 μg/kg, respectively. Acrylamide has not been found in raw foods or foods which have been cooked by boiling. Its content in all other foods had concentrations in the range, 65-3,640 μg/kg. The relationship between acrylamide levels and precursor concentration as well as the health implications of our findings are discussed. Copyright © 2012 Elsevier Ltd. All rights reserved.

  2. Precise pooling and dispensing of microfluidic droplets towards micro- to macro-world interfacing

    PubMed Central

    Brouzes, Eric; Carniol, April; Bakowski, Tomasz; Strey, Helmut H.

    2014-01-01

    Droplet microfluidics possesses unique properties such as the ability to carry out multiple independent reactions without dispersion of samples in microchannels. We seek to extend the use of droplet microfluidics to a new range of applications by enabling its integration into workflows based on traditional technologies, such as microtiter plates. Our strategy consists in developing a novel method to manipulate, pool and deliver a precise number of microfluidic droplets. To this aim, we present a basic module that combines droplet trapping with an on-chip valve. We quantitatively analyzed the trapping efficiency of the basic module in order to optimize its design. We also demonstrate the integration of the basic module into a multiplex device that can deliver 8 droplets at every cycle. This device will have a great impact in low throughput droplet applications that necessitate interfacing with macroscale technologies. The micro- to macro- interface is particularly critical in microfluidic applications that aim at sample preparation and has not been rigorously addressed in this context. PMID:25485102

  3. The SHOLO mill: return on investment versus mill design

    Treesearch

    Hugh W. Reynolds; Charles J. Gatchell; Charles J. Gatchell

    1971-01-01

    The newly developed SHOLO (from SHOrt Log) process can be used to convert low-grade hardwood logs into parts for standard warehouse pallets and pulp chips. Should you build a SHOLO mill? This paper has been prepared to help you decide.

  4. A CMOS IC-based multisite measuring system for stimulation and recording in neural preparations in vitro

    PubMed Central

    Tateno, Takashi; Nishikawa, Jun

    2014-01-01

    In this report, we describe the system integration of a complementary metal oxide semiconductor (CMOS) integrated circuit (IC) chip, capable of both stimulation and recording of neurons or neural tissues, to investigate electrical signal propagation within cellular networks in vitro. The overall system consisted of three major subunits: a 5.0 × 5.0 mm CMOS IC chip, a reconfigurable logic device (field-programmable gate array, FPGA), and a PC. To test the system, microelectrode arrays (MEAs) were used to extracellularly measure the activity of cultured rat cortical neurons and mouse cortical slices. The MEA had 64 bidirectional (stimulation and recording) electrodes. In addition, the CMOS IC chip was equipped with dedicated analog filters, amplification stages, and a stimulation buffer. Signals from the electrodes were sampled at 15.6 kHz with 16-bit resolution. The measured input-referred circuitry noise was 10.1 μ V root mean square (10 Hz to 100 kHz), which allowed reliable detection of neural signals ranging from several millivolts down to approximately 33 μ Vpp. Experiments were performed involving the stimulation of neurons with several spatiotemporal patterns and the recording of the triggered activity. An advantage over current MEAs, as demonstrated by our experiments, includes the ability to stimulate (voltage stimulation, 5-bit resolution) spatiotemporal patterns in arbitrary subsets of electrodes. Furthermore, the fast stimulation reset mechanism allowed us to record neuronal signals from a stimulating electrode around 3 ms after stimulation. We demonstrate that the system can be directly applied to, for example, auditory neural prostheses in conjunction with an acoustic sensor and a sound processing system. PMID:25346683

  5. Target-responsive DNAzyme cross-linked hydrogel for visual quantitative detection of lead.

    PubMed

    Huang, Yishun; Ma, Yanli; Chen, Yahong; Wu, Xuemeng; Fang, Luting; Zhu, Zhi; Yang, Chaoyong James

    2014-11-18

    Because of the severe health risks associated with lead pollution, rapid, sensitive, and portable detection of low levels of Pb(2+) in biological and environmental samples is of great importance. In this work, a Pb(2+)-responsive hydrogel was prepared using a DNAzyme and its substrate as cross-linker for rapid, sensitive, portable, and quantitative detection of Pb(2+). Gold nanoparticles (AuNPs) were first encapsulated in the hydrogel as an indicator for colorimetric analysis. In the absence of lead, the DNAzyme is inactive, and the substrate cross-linker maintains the hydrogel in the gel form. In contrast, the presence of lead activates the DNAzyme to cleave the substrate, decreasing the cross-linking density of the hydrogel and resulting in dissolution of the hydrogel and release of AuNPs for visual detection. As low as 10 nM Pb(2+) can be detected by the naked eye. Furthermore, to realize quantitative visual detection, a volumetric bar-chart chip (V-chip) was used for quantitative readout of the hydrogel system by replacing AuNPs with gold-platinum core-shell nanoparticles (Au@PtNPs). The Au@PtNPs released from the hydrogel upon target activation can efficiently catalyze the decomposition of H2O2 to generate a large volume of O2. The gas pressure moves an ink bar in the V-chip for portable visual quantitative detection of lead with a detection limit less than 5 nM. The device was able to detect lead in digested blood with excellent accuracy. The method developed can be used for portable lead quantitation in many applications. Furthermore, the method can be further extended to portable visual quantitative detection of a variety of targets by replacing the lead-responsive DNAzyme with other DNAzymes.

  6. Microbiological and physicochemical factors affecting Aspergillus section Flavi incidence in Cavendish banana (Musa cavendishii) chips production in Southern Philippines.

    PubMed

    Sales, A C; Azanza, P V; Yoshizawa, T

    2005-01-01

    Microbiological and physicochemical factors affecting the incidence of Aspergillus section Flavi in dried Cavendish banana (Musa cavendishii) chips production in Southern Philippines were examined. The average counts of Aspergillus section Flavi (AFC) in fresh and dried Cavendish bananas from 10 production batches of the Philippine Agro-Industrial Development Cooperative in Davao del Norte, Southern Philippines were 1.2 x 10(2) and 1.6 x 10(2) cfu/g, respectively. Isolates from both samples were identified to be Aspergillus flavus based on spore type and conidial structure of isolates. An increasing trend in the AFC of Cavendish bananas was observed during dried banana chips processing. Variability in the AFC between production batches was attributed to differences in aerobic and fungal populations and physicochemical characteristics of the fruits, peel damage of the raw materials, concentration of AFC in the air and food-contact surfaces of the production area, and temperature and relative humidity (RH) conditions of the environment during production and storage. Physicochemical characteristics of Cavendish bananas from the receipt of raw materials up to the first day of drying were within the reported range of values allowing growth and toxin production by aflatoxigenic fungi. Air-borne AFC varied depending on the section of the production area examined. The close proximity of the waste disposal area from the production operation to the preparation, drying and storage areas suggests that cross-contamination, probably air-borne or insect-borne was a likely occurrence. The hands of workers were also identified as AFC sources. Results of this study highlight the need for the development of strategies to control aflatoxigenic fungi and aflatoxin contamination in Philippine dried Cavendish bananas.

  7. A CMOS IC-based multisite measuring system for stimulation and recording in neural preparations in vitro.

    PubMed

    Tateno, Takashi; Nishikawa, Jun

    2014-01-01

    In this report, we describe the system integration of a complementary metal oxide semiconductor (CMOS) integrated circuit (IC) chip, capable of both stimulation and recording of neurons or neural tissues, to investigate electrical signal propagation within cellular networks in vitro. The overall system consisted of three major subunits: a 5.0 × 5.0 mm CMOS IC chip, a reconfigurable logic device (field-programmable gate array, FPGA), and a PC. To test the system, microelectrode arrays (MEAs) were used to extracellularly measure the activity of cultured rat cortical neurons and mouse cortical slices. The MEA had 64 bidirectional (stimulation and recording) electrodes. In addition, the CMOS IC chip was equipped with dedicated analog filters, amplification stages, and a stimulation buffer. Signals from the electrodes were sampled at 15.6 kHz with 16-bit resolution. The measured input-referred circuitry noise was 10.1 μ V root mean square (10 Hz to 100 kHz), which allowed reliable detection of neural signals ranging from several millivolts down to approximately 33 μ Vpp. Experiments were performed involving the stimulation of neurons with several spatiotemporal patterns and the recording of the triggered activity. An advantage over current MEAs, as demonstrated by our experiments, includes the ability to stimulate (voltage stimulation, 5-bit resolution) spatiotemporal patterns in arbitrary subsets of electrodes. Furthermore, the fast stimulation reset mechanism allowed us to record neuronal signals from a stimulating electrode around 3 ms after stimulation. We demonstrate that the system can be directly applied to, for example, auditory neural prostheses in conjunction with an acoustic sensor and a sound processing system.

  8. Atom-chip based quantum gravimetry for the precise determination of absolute local gravity

    NASA Astrophysics Data System (ADS)

    Abend, S.

    2015-12-01

    We present a novel technique for the precise measurement of absolute local gravity based on cold atom interferometry. Atom interferometry utilizes the interference of matter waves interrogated by laser light to read out inertial forces. Today's generation of these devices typically operate with test mass samples, that consists of ensembles of laser cooled atoms. Their performance is limited by the velocity spread and finite-size of the test masses that impose systematic uncertainties at the level of a few μGal. Rather than laser cooled atoms we employ quantum degenerate ensembles, so called Bose-Einstein condensates, as ultra-sensitive probes for gravity. These sources offer unique properties in temperature as well as in ensemble size that will allow to overcome the current limitations with the next generation of sensors. Furthermore, atom-chip technologies offer the possibility to generate Bose-Einstein condensates in a fast and reliable way. We show a lab-based prototype that uses the atom-chip itself to retro-reflect the interrogation laser and thus serving as inertial reference inside the vacuum. With this setup it is possible to demonstrate all necessary steps to measure gravity, including the preparation of the source, spanning an interferometer as well as the detection of the output signal, within an area of 1 cm3 right below the atom-chip and to analyze relevant systematic effects. In the framework of the center of excellence geoQ a next generation device is under construction at the Institut für Quantenoptik, that will allow for in-field measurements. This device will feature a state-of-the-art atom-chip source with a high-flux of ultra-cold atoms at a repetition rate of 1-2 Hz. In cooperation with the Müller group at the Institut für Erdmessung the sensor will be characterized in the laboratory first, to be ultimately employed in campaigns to measure the Fennoscandian uplift at the level of 1 μGal. The presented work is part of the center of excellence geoQ (SFB 1128), funded by the Deutsche Forschungsgemeinschaft (DFG). This work is supported by the German Space Agency (DLR) with funds provided by the Federal Ministry for Economic Affairs and Energy (BMWi) due to an enactment of the German Bundestag under grant numbers DLR 50 1131-1137 (QUANTUS-III).

  9. Identification of a novel umami compound in potatoes and potato chips.

    PubMed

    Zhang, Liyun; Peterson, Devin G

    2018-02-01

    The influence of frying time on the taste profile of potato chips was characterized. Direct comparison of isolates from potato chip samples fried for 170s and 210s indicated longer frying time increased the perceived umami intensity and decreased the sour intensity. The compounds responsible for the greater umami intensity were identified as monosodium l-pyroglutamate (l-MSpG) and monosodium d-pyroglutamate (d-MSpG). The reduction in sour intensity was attributed to the degradation of d-chlorogenic acid. MSpGs were endogenous in raw potatoes and also thermally generated from glutamic acid during frying. Taste recombination studies further confirmed the contribution of both compounds to the umami character of potato chips. Furthermore, time-intensity taste analysis revealed that topical addition of both l- and d-MSpG enhanced the perceived intensity of the umami taste and the overall flavor characteristic of the potato chips. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Autogenous bone chips: influence of a new piezoelectric device (Piezosurgery) on chip morphology, cell viability and differentiation.

    PubMed

    Chiriac, G; Herten, M; Schwarz, F; Rothamel, D; Becker, J

    2005-09-01

    The aim of the present study was to investigate the influence of a new piezoelectric device, designed for harvesting autogenous bone chips from intra-oral sites, on chip morphology, cell viability and differentiation. A total of 69 samples of cortical bone chips were randomly gained by either (1) a piezoelectric device (PS), or (2) conventional rotating drills (RD). Shape and size of the bone chips were compared by means of morphometrical analysis. Outgrowing osteoblasts were identified by means of alkaline phosphatase activity (AP), immunhistochemical staining for osteocalcin (OC) synthesis and reverse transcriptase-polymerase chain reaction phenotyping. In 88.9% of the RD and 87.9% of the PS specimens, an outgrowth of adherent cells nearby the bone chips was observed after 6-19 days. Confluence of cells was reached after 4 weeks. Positive staining for AP and OC identified the cells as osteoblasts. The morphometrical analysis revealed a statistically significant more voluminous size of the particles collected with PS than RD. Within the limits of the present study, it may be concluded that both the harvesting methods are not different from each other concerning their detrimental effect on viability and differentiation of cells growing out of autogenous bone chips derived from intra-oral cortical sites.

  11. Sparsity-Based Super Resolution for SEM Images.

    PubMed

    Tsiper, Shahar; Dicker, Or; Kaizerman, Idan; Zohar, Zeev; Segev, Mordechai; Eldar, Yonina C

    2017-09-13

    The scanning electron microscope (SEM) is an electron microscope that produces an image of a sample by scanning it with a focused beam of electrons. The electrons interact with the atoms in the sample, which emit secondary electrons that contain information about the surface topography and composition. The sample is scanned by the electron beam point by point, until an image of the surface is formed. Since its invention in 1942, the capabilities of SEMs have become paramount in the discovery and understanding of the nanometer world, and today it is extensively used for both research and in industry. In principle, SEMs can achieve resolution better than one nanometer. However, for many applications, working at subnanometer resolution implies an exceedingly large number of scanning points. For exactly this reason, the SEM diagnostics of microelectronic chips is performed either at high resolution (HR) over a small area or at low resolution (LR) while capturing a larger portion of the chip. Here, we employ sparse coding and dictionary learning to algorithmically enhance low-resolution SEM images of microelectronic chips-up to the level of the HR images acquired by slow SEM scans, while considerably reducing the noise. Our methodology consists of two steps: an offline stage of learning a joint dictionary from a sequence of LR and HR images of the same region in the chip, followed by a fast-online super-resolution step where the resolution of a new LR image is enhanced. We provide several examples with typical chips used in the microelectronics industry, as well as a statistical study on arbitrary images with characteristic structural features. Conceptually, our method works well when the images have similar characteristics, as microelectronics chips do. This work demonstrates that employing sparsity concepts can greatly improve the performance of SEM, thereby considerably increasing the scanning throughput without compromising on analysis quality and resolution.

  12. Gene expression variations during Drosophila metamorphosis in real and simulated gravity

    NASA Astrophysics Data System (ADS)

    Marco, R.; Leandro-García, L. J.; Benguría, A.; Herranz, R.; Zeballos, A.; Gassert, G.; van Loon, J. J.; Medina, F. J.

    Establishing the extent and significance of the effects of the exposure to microgravity of complex living organisms is a critical piece of information if the long-term exploration of near-by planets involving human beings is going to take place in the Future As a first step in this direction we have started to look into the patterns of gene expression during Drosophila development in real and simulated microgravity using microarray analysis of mRNA isolated from samples exposed to different environmental conditions In these experiments we used Affymetrix chips version 1 0 containing probes for more than 14 000 genes almost the complete Drosophila genome 55 of which are tagged with some molecular or functional designation while 45 are still waiting to be identified in functional terms The real microgravity exposure was imposed on the samples during the crew exchanging Soyuz 8 Mission to the ISS in October 2003 when after 11 days in Microgravity the Spanish-born astronaut Pedro Duque returned in the Soyuz 7 capsule carrying the experiments prepared by our Team Due to the constraints in the current ISS experiments in these Missions we limited the stages explored in our experiment to the developmental processes occurring during Drosophila metamorphosis As the experimental conditions at the launch site Baikonour were fairly limited we prepared the experiment in Madrid Toulouse and transp o rted the samples at 15 C in a temperature controlled container to slow down the developmental process a

  13. Fractionation of Exosomes and DNA using Size-Based Separation at the Nanoscale

    NASA Astrophysics Data System (ADS)

    Wunsch, Benjamin; Smith, Joshua; Wang, Chao; Gifford, Stacey; Brink, Markus; Bruce, Robert; Solovitzky, Gustavo; Austin, Robert; Astier, Yann

    Exosomes, a key target of ``liquid biopsies'', are nano-vesicles found in nearly all biological fluids. Exosomes are secreted by eukaryotic and prokaryotic cells alike, and contain information about their originating cells, including surface proteins, cytoplasmic proteins, and nucleic acids. One challenge in studying exosome morphology is the difficulty of sorting exosomes by size and surface markers. Common separation techniques for exosomes include ultracentrifugation and ultrafiltration, for preparation of large volume samples, but these techniques often show contamination and significant heterogeneity between preparations. To date, deterministic lateral displacement (DLD) pillar arrays in silicon have proven an efficient technology to sort, separate, and enrich micron-scale particles including human parasites, eukaryotic cells, blood cells, and circulating tumor cells in blood; however, the DLD technology has never been translated to the true nanoscale, where it could function on bio-colloids such as exosomes. We have fabricated nanoscale DLD (nanoDLD) arrays capable of rapidly sorting colloids down to 20 nm in continuous flow, and demonstrated size sorting of individual exosome vesicles and dsDNA polymers, opening the potential for on-chip biomolecule separation and diagnosti

  14. Differentiation of the seven major lyssavirus species by oligonucleotide microarray.

    PubMed

    Xi, Jin; Guo, Huancheng; Feng, Ye; Xu, Yunbin; Shao, Mingfu; Su, Nan; Wan, Jiayu; Li, Jiping; Tu, Changchun

    2012-03-01

    An oligonucleotide microarray, LyssaChip, has been developed and verified as a highly specific diagnostic tool for differentiation of the 7 major lyssavirus species. As with conventional typing microarray methods, the LyssaChip relies on sequence differences in the 371-nucleotide region coding for the nucleoprotein. This region was amplified using nested reverse transcription-PCR primers that bind to the 7 major lyssaviruses. The LyssaChip includes 57 pairs of species typing and corresponding control oligonucleotide probes (oligoprobes) immobilized on glass slides, and it can analyze 12 samples on a single slide within 8 h. Analysis of 111 clinical brain specimens (65 from animals with suspected rabies submitted to the laboratory and 46 of butchered dog brain tissues collected from restaurants) showed that the chip method was 100% sensitive and highly consistent with the "gold standard," a fluorescent antibody test (FAT). The chip method could detect rabies virus in highly decayed brain tissues, whereas the FAT did not, and therefore the chip test may be more applicable to highly decayed brain tissues than the FAT. LyssaChip may provide a convenient and inexpensive alternative for diagnosis and differentiation of rabies and rabies-related diseases.

  15. DNA Extraction by Isotachophoresis in a Microfluidic Channel

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Stephenson, S J

    Biological assays have many applications. For example, forensics personnel and medical professionals use these tests to diagnose diseases and track their progression or identify pathogens and the host response to them. One limitation of these tests, however, is that most of them target only one piece of the sample - such as bacterial DNA - and other components (e.g. host genomic DNA) get in the way, even though they may be useful for different tests. To address this problem, it would be useful to extract several different substances from a complex biological sample - such as blood - in anmore » inexpensive and efficient manner. This summer, I worked with Maxim Shusteff at Lawrence Livermore National Lab on the Rapid Automated Sample Prep project. The goal of the project is to solve the aforementioned problem by creating a system that uses a series of different extraction methods to extract cells, bacteria, and DNA from a complex biological sample. Biological assays can then be run on purified output samples. In this device, an operator could input a complex sample such as blood or saliva, and would receive separate outputs of cells, bacteria, viruses, and DNA. I had the opportunity to work this summer with isotachophoresis (ITP), a technique that can be used to extract nucleic acids from a sample. This technique is intended to be the last stage of the purification device. Isotachophoresis separates particles based on different electrophoretic mobilities. This technique is convenient for out application because free solution DNA mobility is approximately equal for DNA longer than 300 base pairs in length. The sample of interest - in our case DNA - is fed into the chip with streams of leading electrolyte (LE) and trailing electrolyte (TE). When an electric field is applied, the species migrate based on their electrophoretic mobilities. Because the ions in the leading electrolyte have a high electrophoretic mobility, they race ahead of the slower sample and trailing electrolyte ions. Conversely, the trailing electrolyte ions have a slow electrophoretic mobility, so they lag behind the sample, thus trapping the species of interest between the LE and TE streams. In a typical isotachophoresis configuration, the electric field is applied in a direction parallel to the direction of flow. The species then form bands that stretch across the width of the channel. A major limitation of that approach is that only a finite amount of sample can be processed at once, and the sample must be processed in batches. For our purposes, a form of free-flow isotachophoresis is more convenient, where the DNA forms a band parallel to the edges of the channel. To achieve this, in our chip, the electric field is applied transversely. This creates a force perpendicular to the direction of flow, which causes the different ions to migrate across the flow direction. Because the mobility of the DNA is between the mobility of the leading and the trailing electrolyte, the DNA is focused in a tight band near the center of the channel. The stream of DNA can then be directed to a different output to produce a highly concentrated outlet stream without batch processing. One hurdle that must be overcome for successful ITP is isolating the electrochemical reactions that result from the application of high voltage for the actual process of isotachophoresis. The electrochemical reactions that occur around metal electrodes produce bubbles and pH changes that are detrimental to successful ITP. The design of the chips we use incorporates polyacrylamide gels to serve as electrodes along the central channel. For our design, the metal electrodes are located away from the chip, and high conductivity buffer streams carry the potential to the chip, functioning as a 'liquid electrode.' The stream then runs alongside a gel barrier. The gel electrode permits ion transfer while simultaneously isolating the separation chamber from any contaminants in the outer, 'liquid electrode' streams. The difference in potential from one side of the chip to the other creates an electric field. This field traverses the inner, separation channel, containing the leading electrolyte, the trailing electrolyte, and the sample of interest (DNA). To increase the ease of use of the chips, a newer chip design has been fabricated. This design has wire electrodes integrated on the chip, rather than elsewhere. To keep the pH changes and bubbling isolated from the separation channel, the chip contains deeper wells near the electrodes so that the flowing buffer can wash away any gases that form around the electrode. This design is significantly more compact because it eliminates the cumbersome electrode boxes. Eliminating the electrode boxes also decreases the required voltage, making the experiments safer. This happens because when the 'liquid electrode' streams travel through small diameter tubing, they lose much of their voltage due to the electrical resistance of the fluid in the tubing.« less

  16. Electrochemical behavior of high performance on-chip porous carbon films for micro-supercapacitors applications in organic electrolytes

    NASA Astrophysics Data System (ADS)

    Brousse, K.; Huang, P.; Pinaud, S.; Respaud, M.; Daffos, B.; Chaudret, B.; Lethien, C.; Taberna, P. L.; Simon, P.

    2016-10-01

    Carbide derived carbons (CDCs) are promising materials for preparing integrated micro-supercapacitors, as on-chip CDC films are prepared via a process fully compatible with current silicon-based device technology. These films show good adherence on the substrate and high capacitance thanks to their unique nanoporous structure which can be fine-tuned by adjusting the synthesis parameters during chlorination of the metallic carbide precursor. The carbon porosity is mostly related to the synthesis temperature whereas the thickness of the films depends on the chlorination duration. Increasing the pore size allows the adsorption of large solvated ions from organic electrolytes and leads to higher energy densities. Here, we investigated the electrochemical behavior and performance of on-chip TiC-CDC in ionic liquid solvent mixtures of 1-ethyl-3-methylimidazolium tetrafluoroborate (EMIBF4) diluted in either acetonitrile or propylene carbonate via cyclic voltammetry and electrochemical impedance spectroscopy. Thin CDC films exhibited typical capacitive signature and achieved 169 F cm-3 in both electrolytes; 65% of the capacitance was still delivered at 1 V s-1. While increasing the thickness of the films, EMI+ transport limitation was observed in more viscous PC-based electrolyte. Nevertheless, the energy density reached 90 μW h cm-2 in 2M EMIBF4/ACN, confirming the interest of these CDC films for micro-supercapacitors applications.

  17. Integrated Microfluidic Gas Sensors for Water Monitoring

    NASA Technical Reports Server (NTRS)

    Zhu, L.; Sniadecki, N.; DeVoe, D. L.; Beamesderfer, M.; Semancik, S.; DeVoe, D. L.

    2003-01-01

    A silicon-based microhotplate tin oxide (SnO2) gas sensor integrated into a polymer-based microfluidic system for monitoring of contaminants in water systems is presented. This device is designed to sample a water source, control the sample vapor pressure within a microchannel using integrated resistive heaters, and direct the vapor past the integrated gas sensor for analysis. The sensor platform takes advantage of novel technology allowing direct integration of discrete silicon chips into a larger polymer microfluidic substrate, including seamless fluidic and electrical interconnects between the substrate and silicon chip.

  18. An acoustic on-chip goniometer for room temperature macromolecular crystallography.

    PubMed

    Burton, C G; Axford, D; Edwards, A M J; Gildea, R J; Morris, R H; Newton, M I; Orville, A M; Prince, M; Topham, P D; Docker, P T

    2017-12-05

    This paper describes the design, development and successful use of an on-chip goniometer for room-temperature macromolecular crystallography via acoustically induced rotations. We present for the first time a low cost, rate-tunable, acoustic actuator for gradual in-fluid sample reorientation about varying axes and its utilisation for protein structure determination on a synchrotron beamline. The device enables the efficient collection of diffraction data via a rotation method from a sample within a surface confined droplet. This method facilitates efficient macromolecular structural data acquisition in fluid environments for dynamical studies.

  19. Multi-spot porous silicon chip prepared from asymmetric electrochemical etching for human immunoglobin G sensor.

    PubMed

    Um, Sungyong; Cho, Bomin; Woo, Hee-Gweon; Sohn, Honglae

    2011-08-01

    Multi-spot porous silicon (MSPS)-based optical biosensor was developed to specify the biomolecules. MSPS chip was generated by an electrochemical etching of silicon wafer using an asymmetric electrode configuration in aqueous ethanolic HF solution and constituted with nine arrayed porous silicon. MSPS prepared from anisotropic etching conditions displayed the Fabry-Pérot fringe patterns which varied spatially across the porous silicon (PS). Each spot displayed different reflection resonances and different pore characteristics as a function of the lateral distance from the Pt counter electrode. The sensor system consists of the 3 x 3 spot array of porous silicon modified with Protein A. The system was probed with various fragments of an aqueous Human Immunoglobin G (Ig G) analyte. The sensor operated by measurement of the reflection patterns in the white light reflection spectrum of MSPS. Molecular binding and specificity was detected as a shift in wavelength of these Fabry-Pérot fringe patterns.

  20. Photoluminescence of Alq3 - and Tb-activated aluminium-tris(8-hydroxyquinoline) complex for blue chip-excited OLEDs.

    PubMed

    Yawalkar, P W; Dhoble, S J; Thejo Kalyani, N; Atram, R G; Kokode, N S

    2013-01-01

    The tris(8-hydroxyquinoline)-aluminium complex is the most important and widely studied as electron transporting and green light emitting material. Alq(3) and Tb(x) Al((1-x)) q(3) have been synthesized (where x = 0.1, 0.3, 0.5, 0.7 and 0.9) and blended films of Alq(3) and Tb(x) Al((1-x)) q(3) with PMMA and PS at different percentage weight (wt%) concentrations (e.g., 0.1, 1, 5, 10, 25 and 50 wt%) have been prepared. The synthesized materials and their blended thin films have been characterized by a photoluminescence (PL) technique; the synthesis and PL characterization are reported in this paper. The synthesized metal complex shows bright emission of green light with blue light excitation (440 nm) and the prepared Tb(x) Al((1-x)) q(3) phosphor may be applicable in blue chip-excited OLEDs for the newly developed wallpaper lighting technology. Copyright © 2012 John Wiley & Sons, Ltd.

  1. Investigation of electromigration behavior in lead-free flip chip solder bumps

    NASA Astrophysics Data System (ADS)

    Kalkundri, Kaustubh Jayant

    Packaging technology has also evolved over time in an effort to keep pace with the demanding requirements. Wirebond and flip chip packaging technologies have become extremely versatile and ubiquitous in catering to myriad applications due to their inherent potential. This research is restricted strictly to flip chip technology. This technology incorporates a process in which the bare chip is turned upside down, i.e., active face down, and is bonded through the I/O to the substrate, hence called flip chip. A solder interconnect that provides electrical connection between the chip and substrate is bumped on a processed silicon wafer prior to dicing for die-attach. The assembly is then reflow-soldered followed by the underfill process to provide the required encapsulation. The demand for smaller and lighter products has increased the number of I/Os without increasing the package sizes, thereby drastically reducing the size of the flip chip solder bumps and their pitch. Reliability assessment and verification of these devices has gained tremendous importance due to their shrinking size. To add to the complexity, changing material sets that are results of recently enacted lead-free solder legislations have raised some compatibility issues that are already being researched. In addition to materials and process related flip chip challenges such as solder-flux compatibility, Coefficient of Thermal Expansion (CTE) mismatch, underfill-flux compatibility and thermal management, flip chip packages are vulnerable to a comparatively newer challenge, namely electromigration observed in solder bumps. It is interesting to note that electromigration has come to the forefront of challenges only recently. It has been exacerbated by the reduction in bump cross-section due to the seemingly continuous shrinking in package size over time. The focus of this research was to understand the overall electromigration behavior in lead-free (SnAg) flip chip solder bumps. The objectives of the research were to comprehend the physics of failure mechanism in electromigration for lead-free solder bumps assembled in a flip chip ceramic package having thick copper under bump metallization and to estimate the unknown critical material parameters from Black's equation that describe failure due to electromigration. In addition, the intent was to verify the 'use condition reliability' by extrapolation from experimental conditions. The methodology adopted for this research was comprised of accelerated electromigration tests on SnAg flip chip solder bumps assembled on ceramic substrate with a thick copper under bump metallization. The experimental approach was comprised of elaborate measurement of the temperature of each sample by separate metallization resistance exhibiting positive resistance characteristics to overcome the variation in Joule heating. After conducting the constant current experiments and analyzing the failed samples, it was found that the primary electromigration failure mode observed was the dissolution of the thick copper under bump metallization in the solder, leading to a change in resistance. The lifetime data obtained from different experiments was solved simultaneously using a multiple regression approach to yield the unknown Black's equation parameters of current density exponent and activation energy. In addition to the implementation of a systematic failure analysis and data analysis procedure, it was also deduced that thermomigration due to the temperature gradient across the chip does impact the overall electromigration behavior. This research and the obtained results were significant in bridging the gap for an overall understanding of this critical failure mode observed in flip chip solder bumps. The measurement of each individual sample temperature instead of an average temperature enabled an accurate analysis for predicting the 'use condition reliability' of a comparable product. The obtained results and the conclusions can be used as potential inputs in future designs and newer generations of flip chip devices that might undergo aggressive scaling. This will enable these devices to retain their functionality during their intended useful life with minimal threat of failure due to the potent issue of electromigration. (Abstract shortened by UMI.)

  2. Preparation and characterization of maghemite nanoparticles from mild steel for magnetically guided drug therapy.

    PubMed

    Kumar, Nitesh; Kulkarni, Kaustubh; Behera, Laxmidhar; Verma, Vivek

    2017-08-01

    Maghemite (γ-Fe 2 O 3 ) nanoparticles for therapeutic applications are prepared from mild steel but the existing synthesis technique is very cumbersome. The entire process takes around 100 days with multiple steps which lack proper understanding. In the current work, maghemite nanoparticles of cuboidal and spheroidal morphologies were prepared from mild steel chips by a novel cost effective oil reduction technique for magnetically guided intravascular drug delivery. The technique developed in this work yields isometric sized γ-Fe 2 O 3 nanoparticles in 6 h with higher saturation magnetization as compared to the existing similar solid state synthesis route. Mass and heat flow kinetics during the heating and quenching steps were studied with the help of Finite element simulations. Qualitative and quantitative analysis of the γ-Fe 2 O 3 phase is performed with the help of x-ray diffraction, transmission electron microscope and x-ray photoelectron spectroscopy. Mechanism for the α-Fe 2 O 3 (haematite) to γ-Fe 2 O 3 (maghemite) phase evolution during the synthesis process is also investigated. Maghemite (γ-Fe 2 O 3 ) nanoparticles were prepared bya novel cost effective oil reduction technique as mentioned below in the figure. The raw materials included mild steel chips which is one of the most abundant engineering materials. These particles can be used as ideal nanocarriers for targeted drug delivery through the vascular network.

  3. Sample flow switching techniques on microfluidic chips.

    PubMed

    Pan, Yu-Jen; Lin, Jin-Jie; Luo, Win-Jet; Yang, Ruey-Jen

    2006-02-15

    This paper presents an experimental investigation into electrokinetically focused flow injection for bio-analytical applications. A novel microfluidic device for microfluidic sample handling is presented. The microfluidic chip is fabricated on glass substrates using conventional photolithographic and chemical etching processes and is bonded using a high-temperature fusion method. The proposed valve-less device is capable not only of directing a single sample flow to a specified output port, but also of driving multiple samples to separate outlet channels or even to a single outlet to facilitate sample mixing. The experimental results confirm that the sample flow can be electrokinetically pre-focused into a narrow stream and guided to the desired outlet port by means of a simple control voltage model. The microchip presented within this paper has considerable potential for use in a variety of applications, including high-throughput chemical analysis, cell fusion, fraction collection, sample mixing, and many other applications within the micro-total-analysis systems field.

  4. Coping with Health Problems: Developing a Reliable and Valid Multidimensional Measure.

    ERIC Educational Resources Information Center

    Endler, Norman S.; Parker, James D. A.; Summerfeldt, Laura J.

    1998-01-01

    A self-report measure, the Coping with Health Injuries and Problems Scale (CHIP), was developed to identify basic coping dimensions for responding to health problems. The CHIP factor structure, established with samples of 532 adults and 598 adults in Canada, is cross-validated with 390 general medical patients and 286 chronic back pain patients.…

  5. [The development of an intelligent four-channel aggregometer].

    PubMed

    Guan, X; Wang, M

    1998-07-01

    The paper introduces the hardware and software design of the instrument. We use 89C52 single-chip computer as the microprocessor to control the amplifier, AD and DA conversion chip to realize the sampling, data process, printout and supervision. The final result is printed out in form of data and aggregation curve from PP40 plotter.

  6. Particleboard made from remediated CCA-treated wood : evaluation of panel properties

    Treesearch

    Carol A. Clausen; S. Nami Kartal; James Muehl

    2001-01-01

    CCA-treated southern yellow pine (SYP) chips were remediated utilizing acid extraction alone, and using acid extraction followed by bioleaching with the metal-tolerant bacterium Bacillus licheniformis CC01. bCleanedc chips were used to make particleboard (PB) with 10 percent urea-formaldehyde (UF) resin, and the PB samples were evaluated for internal bond (IB), modulus...

  7. Microbial community analysis of an Alabama coastal salt marsh impacted by the Deepwater Horizon Oil Spill

    NASA Astrophysics Data System (ADS)

    Beazley, M. J.; Martinez, R.; Rajan, S.; Powell, J.; Piceno, Y.; Tom, L.; Andersen, G. L.; Hazen, T. C.; Van Nostrand, J. D.; Zhou, J.; Mortazavi, B.; Sobecky, P. A.

    2011-12-01

    Microbial community responses of an Alabama coastal salt marsh environment to the Deepwater Horizon oil spill were studied by 16S rRNA (PhyloChip) and functional gene (GeoChip) microarray-based analysis. Oil and tar balls associated with the oil spill arrived along the Alabama coast in June 2010. Marsh and inlet sediment samples collected in June, July, and September 2010 from a salt marsh ecosystem at Point Aux Pines Alabama were analyzed to determine if bacterial community structure changed as a result of oil perturbation. Sediment total petroleum hydrocarbon (TPH) concentrations ranged from below detection to 189 mg kg-1 and were randomly dispersed throughout the salt marsh sediments. Total DNA extracted from sediment and particulates were used for PhyloChip and GeoChip hybridization. A total of 4000 to 8000 operational taxonomic units (OTUs) were detected in marsh and inlet samples. Distinctive changes in the number of detectable OTUs were observed between June, July, and September 2010. Surficial inlet sediments demonstrated a significant increase in the total number of OTUs between June and September that correlated with TPH concentrations. The most significant increases in bacterial abundance were observed in the phyla Actinobacteria, Firmicutes, Gemmatimonadetes, Proteobacteria, and Verrucomicrobia. Bacterial richness in marsh sediments also correlated with TPH concentrations with significant changes primarily in Acidobacteria, Actinobacteria, Firmicutes, Fusobacteria, Nitrospirae, and Proteobacteria. GeoChip microarray analysis detected 5000 to 8300 functional genes in marsh and inlet samples. Surficial inlet sediments demonstrated distinctive increases in the number of detectable genes and gene signal intensities in July samples compared to June. Signal intensities increased (> 1.5-fold) in genes associated with petroleum degradation. Genes related to metal resistance, stress, and carbon cycling also demonstrated increases in oiled sediments. This study demonstrates the value of applying phylogenetic and functional gene microarray technology to characterize the extensive microbial diversity of marsh environments. Moreover, this technology provides significant insight into bacterial community responses to anthropogenic oil events.

  8. Drying of restructured chips made from the old stalks of Asparagus officinalis: impact of different drying methods.

    PubMed

    Liu, Zhenbin; Zhang, Min; Wang, Yuchuan

    2016-06-01

    Old stalks of Asparagus officinalis, which account for one third of the total length of each spear, are always discarded as waste. To make full use of the resource, a kind of restructured Asparagus officinalis chip was made. The effects of pulse-spouted microwave-assisted vacuum drying (PSMVD), microwave-assisted vacuum drying (MVD) and vacuum drying (VD) on texture, color and other quality parameters of restructured chips were then studied to obtain high-quality dried chips. Results indicated that the drying time was significantly affected by drying methods, and PSMVD had much better drying uniformity than MVD. The expansion ratio and crispness of chips increased with increasing microwave power and vacuum degree. The browning reaction of samples in VD was more serious, which was confirmed by the results of color test and electronic nose. The PSMVD drying method showed much better drying uniformity than MVD. The dried chips obtained by PSMVD showed optimal quality and were more readily accepted by consumers. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  9. Development of a Three-Dimensional (3D) Printed Biodegradable Cage to Convert Morselized Corticocancellous Bone Chips into a Structured Cortical Bone Graft.

    PubMed

    Chou, Ying-Chao; Lee, Demei; Chang, Tzu-Min; Hsu, Yung-Heng; Yu, Yi-Hsun; Liu, Shih-Jung; Ueng, Steve Wen-Neng

    2016-04-20

    This study aimed to develop a new biodegradable polymeric cage to convert corticocancellous bone chips into a structured strut graft for treating segmental bone defects. A total of 24 adult New Zealand white rabbits underwent a left femoral segmental bone defect creation. Twelve rabbits in group A underwent three-dimensional (3D) printed cage insertion, corticocancellous chips implantation, and Kirschner-wire (K-wire) fixation, while the other 12 rabbits in group B received bone chips implantation and K-wire fixation only. All rabbits received a one-week activity assessment and the initial image study at postoperative 1 week. The final image study was repeated at postoperative 12 or 24 weeks before the rabbit scarification procedure on schedule. After the animals were sacrificed, both femurs of all the rabbits were prepared for leg length ratios and 3-point bending tests. The rabbits in group A showed an increase of activities during the first week postoperatively and decreased anterior cortical disruptions in the postoperative image assessments. Additionally, higher leg length ratios and 3-point bending strengths demonstrated improved final bony ingrowths within the bone defects for rabbits in group A. In conclusion, through this bone graft converting technique, orthopedic surgeons can treat segmental bone defects by using bone chips but with imitate characters of structured cortical bone graft.

  10. Development of a Three-Dimensional (3D) Printed Biodegradable Cage to Convert Morselized Corticocancellous Bone Chips into a Structured Cortical Bone Graft

    PubMed Central

    Chou, Ying-Chao; Lee, Demei; Chang, Tzu-Min; Hsu, Yung-Heng; Yu, Yi-Hsun; Liu, Shih-Jung; Ueng, Steve Wen-Neng

    2016-01-01

    This study aimed to develop a new biodegradable polymeric cage to convert corticocancellous bone chips into a structured strut graft for treating segmental bone defects. A total of 24 adult New Zealand white rabbits underwent a left femoral segmental bone defect creation. Twelve rabbits in group A underwent three-dimensional (3D) printed cage insertion, corticocancellous chips implantation, and Kirschner-wire (K-wire) fixation, while the other 12 rabbits in group B received bone chips implantation and K-wire fixation only. All rabbits received a one-week activity assessment and the initial image study at postoperative 1 week. The final image study was repeated at postoperative 12 or 24 weeks before the rabbit scarification procedure on schedule. After the animals were sacrificed, both femurs of all the rabbits were prepared for leg length ratios and 3-point bending tests. The rabbits in group A showed an increase of activities during the first week postoperatively and decreased anterior cortical disruptions in the postoperative image assessments. Additionally, higher leg length ratios and 3-point bending strengths demonstrated improved final bony ingrowths within the bone defects for rabbits in group A. In conclusion, through this bone graft converting technique, orthopedic surgeons can treat segmental bone defects by using bone chips but with imitate characters of structured cortical bone graft. PMID:27104525

  11. Dry Matter Losses and Greenhouse Gas Emissions From Outside Storage of Short Rotation Coppice Willow Chip.

    PubMed

    Whittaker, Carly; Yates, Nicola E; Powers, Stephen J; Misselbrook, Tom; Shield, Ian

    This study examined the dry matter losses and the greenhouse gas (GHG) concentrations within two short rotation coppice (SRC) willow wood chip storage heaps. One heap was built on a grassland area (East Midlands) and the other (Rothamsted) on a concrete hard standing. A series of 1- and 3-m probes were embedded in the heaps in order to retrieve gas samples for analysis, and pre-weighed net bags were positioned in the core of the heap to detect dry matter losses. The bagged samples showed dry matter losses of 18 and 19 % in the East Midlands and Rothamsted heaps after 210 and 97 days storage, respectively. The Rothamsted heap showed a whole-heap dry matter loss of 21 %. During this time, the wood chips dried from 54 to 39 % moisture content in the East Midlands heap and 50 to 43 % at Rothamsted. The results from analysing the whole Rothamsted heap indicated an overall loss of 1.5 GJ per tonne stored, although measurements from bagged samples in the core suggested that the chips dried sufficiently to have a minimal energy loss from storage. The process of mixing the heap, however, led to incorporation of wet outer layers and hence the average moisture content was higher in an average sample of chip. After establishment of the heaps, the temperature rose rapidly and this correlated with a peak in carbon dioxide (CO 2 ) concentration within the heap. A peak in methane (CH 4 ) concentration was also detected in both heaps, though more noticeably in the East Midlands heap after around 55 days. In both instances, the peak CH 4 concentration occurred as CO 2 concentrations dropped, suggesting that after an active period of aerobic decomposition in the first 2 months of storage, the conditions in the heap became anaerobic. The results from this study suggest that outside wood chip storage is not an efficient method of storing biomass, though this may be location-specific as there are some studies showing lower dry matter losses. It is necessary to explore other methods of harvesting SRC to minimise losses and optimise land use efficiency. Further research is required to detect whether there are fugitive emissions of CH 4 from wood chip heaps, as this will compromise the net GHG savings from utilising the biomass stored in this way.

  12. Hydroponics on a chip: analysis of the Fe deficient Arabidopsis thylakoid membrane proteome.

    PubMed

    Laganowsky, Arthur; Gómez, Stephen M; Whitelegge, Julian P; Nishio, John N

    2009-04-13

    The model plant Arabidopsis thaliana was used to evaluate the thylakoid membrane proteome under Fe-deficient conditions. Plants were cultivated using a novel hydroponic system, called "hydroponics on a chip", which yields highly reproducible plant tissue samples for physiological analyses, and can be easily used for in vivo stable isotope labeling. The thylakoid membrane proteome, from intact chloroplasts isolated from Fe-sufficient and Fe-deficient plants grown with hydroponics on a chip, was analyzed using liquid chromatography coupled to mass spectrometry. Intact masses of thylakoid membrane proteins were measured, many for the first time, and several proteins were identified with post-translational modifications that were altered by Fe deficiency; for example, the doubly phosphorylated form of the photosystem II oxygen evolving complex, PSBH, increased under Fe-deficiency. Increased levels of photosystem II protein subunit PSBS were detected in the Fe-deficient samples. Antioxidant enzymes, including ascorbate peroxidase and peroxiredoxin Q, were only detected in the Fe-deficient samples. We present the first biochemical evidence that the two major LHC IIb proteins (LHCB1 and LHCB2) may have significantly different functions in the thylakoid membrane. The study illustrates the utility of intact mass proteomics as an indispensable tool for functional genomics. "Hydroponics on a chip" provides the ability to grow A. thaliana under defined conditions that will be useful for systems biology.

  13. Leverage principle of retardation signal in titration of double protein via chip moving reaction boundary electrophoresis.

    PubMed

    Zhang, Liu-Xia; Cao, Yi-Ren; Xiao, Hua; Liu, Xiao-Ping; Liu, Shao-Rong; Meng, Qing-Hua; Fan, Liu-Yin; Cao, Cheng-Xi

    2016-03-15

    In the present work we address a simple, rapid and quantitative analytical method for detection of different proteins present in biological samples. For this, we proposed the model of titration of double protein (TDP) and its relevant leverage theory relied on the retardation signal of chip moving reaction boundary electrophoresis (MRBE). The leverage principle showed that the product of the first protein content and its absolute retardation signal is equal to that of the second protein content and its absolute one. To manifest the model, we achieved theoretical self-evidence for the demonstration of the leverage principle at first. Then relevant experiments were conducted on the TDP-MRBE chip. The results revealed that (i) there was a leverage principle of retardation signal within the TDP of two pure proteins, and (ii) a lever also existed within these two complex protein samples, evidently demonstrating the validity of TDP model and leverage theory in MRBE chip. It was also showed that the proposed technique could provide a rapid and simple quantitative analysis of two protein samples in a mixture. Finally, we successfully applied the developed technique for the quantification of soymilk in adulterated infant formula. The TDP-MRBE opens up a new window for the detection of adulteration ratio of the poor food (milk) in blended high quality one. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Biomonitoring of perfluorinated compounds in a drop of blood.

    PubMed

    Mao, Pan; Wang, Daojing

    2015-06-02

    Biomonitoring of pollutants and their metabolites and derivatives using biofluids provides new opportunities for spatiotemporal assessment of human risks to environmental exposures. Perfluorinated compounds (PFCs) have been used widely in industry and pose significant environmental concerns due to their stability and bioaccumulation in humans and animals. However, current methods for extraction and measurement of PFCs require relatively large volumes (over one hundred microliters) of blood samples, and therefore, are not suitable for frequent blood sampling and longitudinal biomonitoring of PFCs. We have developed a new microassay, enabled by our silicon microfluidic chip platform, for analyzing PFCs in small volumes (less than five microliters) of blood. Our assay integrates on-chip solid-phase extraction (SPE) with online nanoflow liquid chromatography-electrospray ionization-mass spectrometry (nanoLC-ESI-MS) detection. We demonstrated high sample recovery, excellent interday and intraday accuracy and precision, and a limit of detection down to 50 femtogram of PFCs, in one microliter of human plasma. We validated our assay performance using pooled human plasma and NIST SRM 1950 samples. Our microfluidic chip-based assay may enable frequent longitudinal biomonitoring of PFCs and other environmental toxins using a finger prick of blood, thereby providing new insights into their bioaccumulation, bioavailability, and toxicity.

  15. Cytometer on a Chip

    NASA Technical Reports Server (NTRS)

    Fernandez, Salvador M.

    2011-01-01

    A cytometer now under development exploits spatial sorting of sampled cells on a microarray chip followed by use of grating-coupled surface-plasmon-resonance imaging (GCSPRI) to detect the sorted cells. This cytometer on a chip is a prototype of contemplated future miniature cytometers that would be suitable for rapidly identifying pathogens and other cells of interest in both field and laboratory applications and that would be attractive as alternatives to conventional flow cytometers. The basic principle of operation of a conventional flow cytometer requires fluorescent labeling of sampled cells, stringent optical alignment of a laser beam with a narrow orifice, and flow of the cells through the orifice, which is subject to clogging. In contrast, the principle of operation of the present cytometer on a chip does not require fluorescent labeling of cells, stringent optical alignment, or flow through a narrow orifice. The basic principle of operation of the cytometer on a chip also reduces the complexity, mass, and power of the associated laser and detection systems, relative to those needed in conventional flow cytometry. Instead of making cells flow in single file through a narrow flow orifice for sequential interrogation as in conventional flow cytometry, a liquid containing suspended sampled cells is made to flow over the front surface of a microarray chip on which there are many capture spots. Each capture spot is coated with a thin (approximately 50-nm) layer of gold that is, in turn, coated with antibodies that bind to cell-surface molecules characteristic of one the cell species of interest. The multiplicity of capture spots makes it possible to perform rapid, massively parallel analysis of a large cell population. The binding of cells to each capture spot gives rise to a minute change in the index of refraction at the surface of the chip. This change in the index of refraction is what is sensed in GCSPRI, as described briefly below. The identities of the various species in a sample of cells is spatially encoded in the chip by the pattern of capture spots. The number of cells of a particular species is determined from the magnitude of the GCSPRI signal from that spot. GCSPRI as used here can be summarized as follows: The cytometer chip is fabricated with a diffraction grating on its front surface. The chip is illuminated with a light emitting diode (LED) from the front. By proper choice of grating parameters and of the wavelength and the angle of incidence of a laser beam, laser light can be made to be coupled into an electromagnetic mode that resonates with surface plasmons and thus couples light into surface plasmons. Coupling of light into a surface plasmon at a given location reduces the amount of incident light reflected from that location. A change in the index of refraction at the surface of a capture spot gives rise to a change in the resonance condition. Depending on the specific design, the change in the index of refraction could manifest itself as a brightening or darkening, a change in the wavelength needed to excite the plasmon at a given angle of incidence, or a change in the angle of incidence needed to excite the plasmon at a given wavelength. Whereas a multiwavelength laser system with multichannel detection would be needed to detect multiple species in conventional flow cytometry, it suffices to use an LED and a single detector channel in the GCSPRI approach: this contributes significantly to reductions in cost, complexity, size, mass, and power. GCSPRI cytometer chips could be made of plastic and could be mass-produced cheaply by use of molding and other methods adopted from the manufacture of digital video disks. These methods are amenable to a high degree of miniaturization: such additional features as fluidic channels, reaction chambers, and fluid-coupling ports could readily be incorporated into the chips, without incurring substantial additional costs.

  16. Cytometer on a Chip

    NASA Technical Reports Server (NTRS)

    Fernandez, Salvador M.

    2011-01-01

    A cytometer now under development exploits spatial sorting of sampled cells on a microarray chip followed by use of grating-coupled surface-plasmon-resonance imaging (GCSPRI) to detect the sorted cells. This cytometer on a chip is a prototype of contemplated future miniature cytometers that would be suitable for rapidly identifying pathogens and other cells of interest in both field and laboratory applications and that would be attractive as alternatives to conventional flow cytometers. The basic principle of operation of a conventional flow cytometer requires fluorescent labeling of sampled cells, stringent optical alignment of a laser beam with a narrow orifice, and flow of the cells through the orifice, which is subject to clogging. In contrast, the principle of operation of the present cytometer on a chip does not require fluorescent labeling of cells, stringent optical alignment, or flow through a narrow orifice. The basic principle of operation of the cytometer on a chip also reduces the complexity, mass, and power of the associated laser and detection systems, relative to those needed in conventional flow cytometry. Instead of making cells flow in single file through a narrow flow orifice for sequential interrogation as in conventional flow cytometry, a liquid containing suspended sampled cells is made to flow over the front surface of a microarray chip on which there are many capture spots. Each capture spot is coated with a thin (.50-nm) layer of gold that is, in turn, coated with antibodies that bind to cell-surface molecules characteristic of the cell species of interest. The multiplicity of capture spots makes it possible to perform rapid, massively parallel analysis of a large cell population. The binding of cells to each capture spot gives rise to a minute change in the index of refraction at the surface of the chip. This change in the index of refraction is what is sensed in GCSPRI, as described briefly below. The identities of the various species in a sample of cells is spatially encoded in the chip by the pattern of capture spots. The number of cells of a particular species is determined from the magnitude of the GCSPRI signal from that spot. GCSPRI as used here can be summarized as follows: The cytometer chip is fabricated with a diffraction grating on its front surface. The chip is illuminated with a light emitting diode (LED) from the front. By proper choice of grating parameters and of the wavelength and the angle of incidence of a laser beam, laser light can be made to be coupled into an electromagnetic mode that resonates with surface plasmons and thus couples light into surface plasmons. Coupling of light into a surface plasmon at a given location reduces the amount of incident light reflected from that location. A change in the index of refraction at the surface of a capture spot gives rise to a change in the resonance condition. Depending on the specific design, the change in the index of refraction could manifest itself as a brightening or darkening, a change in the wavelength needed to excite the plasmon at a given angle of incidence, or a change in the angle of incidence needed to excite the plasmon at a given wavelength. Whereas a multiwavelength laser system with multichannel detection would be needed to detect multiple species in conventional flow cytometry, it suffices to use an LED and a single detector channel in the GCSPRI approach: this contributes significantly to reductions in cost, complexity, size, mass, and power. GCSPRI cytometer chips could be made of plastic and could be mass-produced cheaply by use of molding and other methods adopted from the manufacture of digital video disks. These methods are amenable to a high degree of miniaturization: such additional features as fluidic channels, reaction chambers, and fluid-coupling ports could readily be incorporated into the chips, without incurring substantial additional costs.

  17. Testing of a Microfluidic Sampling System for High Temperature Electrochemical MC&A

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pereira, Candido; Nichols, Kevin

    2013-11-27

    This report describes the preliminary validation of a high-temperature microfluidic chip system for sampling of electrochemical process salt. Electroanalytical and spectroscopic techniques are attractive candidates for improvement through high-throughput sample analysis via miniaturization. Further, microfluidic chip systems are amenable to micro-scale chemical processing such as rapid, automated sample purification to improve sensor performance. The microfluidic chip was tested to determine the feasibility of the system for high temperature applications and conditions under which microfluidic systems can be used to generate salt droplets at process temperature to support development of material balance and control systems in a used fuel treatment facility.more » In FY13, the project focused on testing a quartz microchip device with molten salts at near process temperatures. The equipment was installed in glove box and tested up to 400°C using commercial thermal transfer fluids as the carrier phase. Preliminary tests were carried out with a low-melting halide salt to initially characterize the properties of this novel liquid-liquid system and to investigate the operating regimes for inducing droplet flow within candidate carrier fluids. Initial results show that the concept is viable for high temperature sampling but further development is required to optimize the system to operate with process relevant molten salts.« less

  18. An effective way to reduce water absorption to terahertz

    NASA Astrophysics Data System (ADS)

    Wu, Yaxiong; Su, Bo; He, Jingsuo; Zhang, Cong; Zhang, Hongfei; Zhang, Shengbo; Zhang, Cunlin

    2018-01-01

    Since many vibrations and rotational levels of biomolecules fall within the THz band, THz spectroscopy can be used to identify biological samples. In addition, most biomolecules need to maintain their biological activity in a liquid environment, but water as polar substance has strong absorption to the THz wave. Thus, it is difficult to detect the sample information in aqueous solution using THz wave. In order to prevent the information of biological samples were masked in the solution, many research methods were used to explore how to reduce the water absorption of terahertz. In this paper, we have developed a real-time chemical methodology through transmission Terahertz time-domain spectroscopy (THz-TDS) system. The material of Zeonor 1020r is used as substrate and cover plate, and PDMS as channel interlayer. The transmission of the empty microfluidic chip is more than 80% in the range of 0.2-2.6 THz by THz-TDS system. Then, experiments were carried out using chips, which were filled with different volumes of 1, 2- propanediol, and it has been proved that the microfluidic chip could reduce the water absorption of terahertz. Finally, in order to further explore the reduction of terahertz to water absorption, we inject different concentrations of electrolyte to the chip. The results show that with the addition of different electrolytes, terahertz transmission line has evident changes. It can be taken into account that the electrolyte has different effects about the hydrogen bonds in the aqueous solution. Some of them can promote water molecules clusters, while others destroy them. Based on the basis of microfluidic chip, the discovery of this phenomenon can provide a way that reduces water absorption of terahertz. This work has laid a solid foundation for the subsequent study in reducing water absorption of terahertz.

  19. Meniscal repair using engineered tissue.

    PubMed

    Peretti, G M; Caruso, E M; Randolph, M A; Zaleske, D J

    2001-03-01

    In this study, devitalized meniscal tissue pre-seeded with viable cultured chondrocytes was used to repair a bucket-handle incision in meniscal tissue transplanted to nude mice. Lamb knee menisci were devitalized by cyclic freezing and thawing. Chips measuring four by two by one-half millimeters were cut from this devitalized tissue to serve as scaffolds. These chips were then cultured either with or without viable allogeneic lamb chondrocytes. From the inner third of the devitalized meniscal tissue, rectangles were also cut approximately 8 x 6 mm. A 4 mm bucket-handle type incision was made in these blocks. The previously prepared chips either with (experimental group) or without viable chondrocytes (control group) were positioned into the incisions and secured with suture. Further control groups included blocks of devitalized menisci with incisions into which no chips were positioned and either closed with suture or left open with no suture. Specimens were transplanted to subcutaneous pouches of nude mice for 14 weeks. After 14 weeks, seven of eight experimental specimens (chips with viable chondrocytes) demonstrated bridging of the incision assessed by gross inspection and manual distraction. All the control groups were markedly different from the experimental group in that the incision remained grossly visible. Histological analysis was consistent with the differences apparent at the gross level. Only the experimental specimens (chips with viable chondrocytes) with gross bridging demonstrated obliteration of the interface between incision and scaffold. None of the control specimens revealed any cells or tissue filling the incision. Tissue engineering using scaffolds and viable cells may have an application in meniscal repair in vivo.

  20. Rapid Detection of Nivalenol and Deoxynivalenol in Wheat Using Surface Plasmon Resonance Immunoassay

    USDA-ARS?s Scientific Manuscript database

    Surface plasmon resonance immunoassay using a monoclonal antibody was developed to measure nivalenol (NIV) and deoxynivalenol (DON) contamination in wheat. A DON-immobilized sensor chip having high sensitivity and stability was prepared, and an SPR detection procedure was developed. The competitiv...

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