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Sample records for ppc microfluidic chips

  1. Purification and preconcentration of genomic DNA from whole cell lysates using photoactivated polycarbonate (PPC) microfluidic chips.

    PubMed

    Witek, Malgorzata A; Llopis, Shawn D; Wheatley, Abigail; McCarley, Robin L; Soper, Steven A

    2006-06-06

    We discuss the use of a photoactivated polycarbonate (PPC) microfluidic chip for the solid-phase, reversible immobilization (SPRI) and purification of genomic DNA (gDNA) from whole cell lysates. The surface of polycarbonate was activated by UV radiation resulting in a photo-oxidation reaction, which produced a channel surface containing carboxylate groups. The gDNA was selectively captured on this photoactivated surface in an immobilization buffer, which consisted of 3% polyethylene glycol, 0.4 M NaCl and 70% ethanol. The methodology reported herein is similar to conventional SPRI in that surface-confined carboxylate groups are used for the selective immobilization of DNA; however, no magnetic beads or a magnetic field are required. As observed by UV spectroscopy, a load of approximately 7.6 +/- 1.6 microg/ml of gDNA was immobilized onto the PPC bed. The recovery of DNA following purification was estimated to be 85 +/- 5%. The immobilization and purification assay using this PPC microchip could be performed within approximately 25 min as follows: (i) DNA immobilization approximately 6 min, (ii) chip washout with ethanol 10 min, and (iii) drying and gDNA desorption approximately 6 min. The PPC microchip could also be used for subsequent assays with no substantial loss in recovery, no observable carryover and no need for 'reactivation' of the PC surface with UV light.

  2. Rapid prototyping of glass microfluidic chips

    NASA Astrophysics Data System (ADS)

    Kotz, Frederik; Plewa, Klaus; Bauer, Werner; Hanemann, Thomas; Waldbaur, Ansgar; Wilhelm, Elisabeth; Neumann, Christiane; Rapp, Bastian E.

    2015-03-01

    In academia the rapid and flexible creation of microfluidic chips is of great importance for microfluidic research. Besides polymers glass is a very important material especially when high chemical and temperature resistance are required. However, glass structuring is a very hazardous process which is not accessible to most members of the microfluidic community. We therefore sought a new method for the rapid and simple creation of transparent microfluidic glass chips by structuring and sintering amorphous silica suspensions. The whole process from a digital mask layout to a microstructured glass sheet can be done within two days. In this paper we show the applicability of this process to fabricate capillary driven microfluidic systems.

  3. Whole-Teflon microfluidic chips.

    PubMed

    Ren, Kangning; Dai, Wen; Zhou, Jianhua; Su, Jing; Wu, Hongkai

    2011-05-17

    Although microfluidics has shown exciting potential, its broad applications are significantly limited by drawbacks of the materials used to make them. In this work, we present a convenient strategy for fabricating whole-Teflon microfluidic chips with integrated valves that show outstanding inertness to various chemicals and extreme resistance against all solvents. Compared with other microfluidic materials [e.g., poly(dimethylsiloxane) (PDMS)] the whole-Teflon chip has a few more advantages, such as no absorption of small molecules, little adsorption of biomolecules onto channel walls, and no leaching of residue molecules from the material bulk into the solution in the channel. Various biological cells have been cultured in the whole-Teflon channel. Adherent cells can attach to the channel bottom, spread, and proliferate well in the channels (with similar proliferation rate to the cells in PDMS channels with the same dimensions). The moderately good gas permeability of the Teflon materials makes it suitable to culture cells inside the microchannels for a long time.

  4. Materials for microfluidic chip fabrication.

    PubMed

    Ren, Kangning; Zhou, Jianhua; Wu, Hongkai

    2013-11-19

    Through manipulating fluids using microfabricated channel and chamber structures, microfluidics is a powerful tool to realize high sensitive, high speed, high throughput, and low cost analysis. In addition, the method can establish a well-controlled microenivroment for manipulating fluids and particles. It also has rapid growing implementations in both sophisticated chemical/biological analysis and low-cost point-of-care assays. Some unique phenomena emerge at the micrometer scale. For example, reactions are completed in a shorter amount of time as the travel distances of mass and heat are relatively small; the flows are usually laminar; and the capillary effect becomes dominant owing to large surface-to-volume ratios. In the meantime, the surface properties of the device material are greatly amplified, which can lead to either unique functions or problems that we would not encounter at the macroscale. Also, each material inherently corresponds with specific microfabrication strategies and certain native properties of the device. Therefore, the material for making the device plays a dominating role in microfluidic technologies. In this Account, we address the evolution of materials used for fabricating microfluidic chips, and discuss the application-oriented pros and cons of different materials. This Account generally follows the order of the materials introduced to microfluidics. Glass and silicon, the first generation microfluidic device materials, are perfect for capillary electrophoresis and solvent-involved applications but expensive for microfabriaction. Elastomers enable low-cost rapid prototyping and high density integration of valves on chip, allowing complicated and parallel fluid manipulation and in-channel cell culture. Plastics, as competitive alternatives to elastomers, are also rapid and inexpensive to microfabricate. Their broad variety provides flexible choices for different needs. For example, some thermosets support in-situ fabrication of

  5. Microfluidic organs-on-chips.

    PubMed

    Bhatia, Sangeeta N; Ingber, Donald E

    2014-08-01

    An organ-on-a-chip is a microfluidic cell culture device created with microchip manufacturing methods that contains continuously perfused chambers inhabited by living cells arranged to simulate tissue- and organ-level physiology. By recapitulating the multicellular architectures, tissue-tissue interfaces, physicochemical microenvironments and vascular perfusion of the body, these devices produce levels of tissue and organ functionality not possible with conventional 2D or 3D culture systems. They also enable high-resolution, real-time imaging and in vitro analysis of biochemical, genetic and metabolic activities of living cells in a functional tissue and organ context. This technology has great potential to advance the study of tissue development, organ physiology and disease etiology. In the context of drug discovery and development, it should be especially valuable for the study of molecular mechanisms of action, prioritization of lead candidates, toxicity testing and biomarker identification.

  6. A Microfluidic Chip for ICPMS Sample Introduction

    PubMed Central

    Verboket, Pascal E.; Borovinskaya, Olga; Meyer, Nicole; Günther, Detlef; Dittrich, Petra S.

    2015-01-01

    This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). The chip produces monodisperse aqueous sample droplets in perfluorohexane (PFH). Size and frequency of the aqueous droplets can be varied in the range of 40 to 60 µm and from 90 to 7,000 Hz, respectively. The droplets are ejected from the chip with a second flow of PFH and remain intact during the ejection. A custom-built desolvation system removes the PFH and transports the droplets into the ICPMS. Here, very stable signals with a narrow intensity distribution can be measured, showing the monodispersity of the droplets. We show that the introduction system can be used to quantitatively determine iron in single bovine red blood cells. In the future, the capabilities of the introduction device can easily be extended by the integration of additional microfluidic modules. PMID:25867751

  7. A microfluidic chip for ICPMS sample introduction.

    PubMed

    Verboket, Pascal E; Borovinskaya, Olga; Meyer, Nicole; Günther, Detlef; Dittrich, Petra S

    2015-03-05

    This protocol discusses the fabrication and usage of a disposable low cost microfluidic chip as sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). The chip produces monodisperse aqueous sample droplets in perfluorohexane (PFH). Size and frequency of the aqueous droplets can be varied in the range of 40 to 60 µm and from 90 to 7,000 Hz, respectively. The droplets are ejected from the chip with a second flow of PFH and remain intact during the ejection. A custom-built desolvation system removes the PFH and transports the droplets into the ICPMS. Here, very stable signals with a narrow intensity distribution can be measured, showing the monodispersity of the droplets. We show that the introduction system can be used to quantitatively determine iron in single bovine red blood cells. In the future, the capabilities of the introduction device can easily be extended by the integration of additional microfluidic modules.

  8. The processing technology of PMMA micro-fluidic chip

    NASA Astrophysics Data System (ADS)

    Mu, Lili; Rong, Li; Guo, Shuheng; Liu, Qiong

    2016-01-01

    In order to enrich the production method of micro-fluidic chip and simplify its processing technology, the paper discussed the double-sided adhesive layer for channel layer, with PMMA (polymethyl methacrylate) for fabrication of microfluidic chip with the cover plate and the bottom plate. Taking 40 mm (long) x 20 mm (wide) x 2.2 mm (thick) liquid drop to separate the microfluidic chip as an example, details the design and machining process of the chip. Experiments show that surface quality is high and processing speed is fast when using this technology to process the chip. Thus, it can realize the mass production of micro fluidic chip.

  9. Microfluidic distillation chip for methanol concentration detection.

    PubMed

    Wang, Yao-Nan; Liu, Chan-Chiung; Yang, Ruey-Jen; Ju, Wei-Jhong; Fu, Lung-Ming

    2016-03-17

    An integrated microfluidic distillation system is proposed for separating a mixed ethanol-methanol-water solution into its constituent components. The microfluidic chip is fabricated using a CO2 laser system and comprises a serpentine channel, a boiling zone, a heating zone, and a cooled collection chamber filled with de-ionized (DI) water. In the proposed device, the ethanol-methanol-water solution is injected into the microfluidic chip and driven through the serpentine channel and into the collection chamber by means of a nitrogen carrier gas. Following the distillation process, the ethanol-methanol vapor flows into the collection chamber and condenses into the DI water. The resulting solution is removed from the collection tank and reacted with a mixed indicator. Finally, the methanol concentration is inversely derived from the absorbance measurements obtained using a spectrophotometer. The experimental results show the proposed microfluidic system achieves an average methanol distillation efficiency of 97%. The practicality of the proposed device is demonstrated by detecting the methanol concentrations of two commercial fruit wines. It is shown that the measured concentration values deviate by no more than 3% from those obtained using a conventional bench top system.

  10. Reagent-loaded plastic microfluidic chips for detecting homocysteine

    NASA Astrophysics Data System (ADS)

    Suk, Ji Won; Jang, Jae-Young; Cho, Jun-Hyeong

    2008-05-01

    This report describes the preliminary study on plastic microfluidic chips with pre-loaded reagents for detecting homocysteine (Hcy). All reagents needed in an Hcy immunoassay were included in a microfluidic chip to remove tedious assay steps. A simple and cost-effective bonding method was developed to realize reagent-loaded microfluidic chips. This technique uses an intermediate layer between two plastic substrates by selectively patterning polydimethylsiloxane (PDMS) on the embossed surface of microchannels and fixing the substrates under pressure. Using this bonding method, the competitive immunoassay for SAH, a converted form of Hcy, was performed without any damage to reagents in chips, and the results showed that the fluorescent signal from antibody antigen binding decreased as the SAH concentration increased. Based on the SAH immunoassay, whole immunoassay steps for Hcy detection were carried out in plastic microfluidic chips with all necessary reagents. These experiments demonstrated the feasibility of the Hcy immunoassay in microfluidic devices.

  11. Hybrid IC / Microfluidic Chips for the Manipulation of Biological Cells

    NASA Astrophysics Data System (ADS)

    Lee, Hakho

    2005-03-01

    A hybrid IC / Microfluidic chip that can manipulate individual biological cells in a fluid with microscopic resolution has been demonstrated. The chip starts with a custom-designed silicon integrated circuit (IC) produced in a foundry using standard processing techniques. A microfluidic chamber is then fabricated on top of the IC to provide a biocompatible environment. The motion of biological cells in the chamber is controlled using a two-dimensional array of micro-scale electromagnets in the IC that generate spatially patterned magnetic fields. A local peak in the magnetic field amplitude will trap a magnetic bead and an attached cell; by moving the peak's location, the bead-bound cell can be moved to any position on the chip surface above the array. By generating multiple peaks, many cells can be moved independently along separate paths, allowing many different manipulations of individual cells. The hybrid IC / Microfluidic chip can be used, for example, to sort cells or to assemble tissue on micrometer length scales. To prove the concept, an IC / Microfluidic chip was fabricated, based on a custom-designed IC that contained a two-dimensional microcoil array with integrated current sources and control circuits. The chip was tested by trapping and moving biological cells tagged with magnetic beads inside the microfluidic chamber over the array. By combining the power of silicon technology with the biocompatibility of microfluidics, IC / Microfluidic chips will make new types of investigations possible in biological and biomedical studies.

  12. Microfluidic chips for protein differential expression profiling.

    PubMed

    Armenta, Jenny M; Dawoud, Abdulilah A; Lazar, Iulia M

    2009-04-01

    Biomarker discovery and screening using novel proteomic technologies is an area that is attracting increased attention in the biomedical community. Early detection of abnormal physiological conditions will be highly beneficial for diagnosing various diseases and increasing survivability rates. Clearly, progress in this area will depend on the development of fast, reliable, and highly sensitive and specific sample bioanalysis methods. Microfluidics has emerged as a technology that could become essential in proteomics research as it enables the integration of all sample preparation, separation, and detection steps, with the added benefit of enhanced sample throughput. The combination of these advantages with the sensitivity and capability of MS detection to deliver precise structural information makes microfluidics-MS a very competitive technology for biomarker discovery. The integration of LC microchip devices with MS detection, and specifically their applicability to biomarker screening applications in MCF-7 breast cancer cellular extracts is reported in this manuscript. Loading approximately 0.1-1 microg of crude protein extract tryptic digest on the chip has typically resulted in the reliable identification of approximately 40-100 proteins. The potential of an LC-ESI-MS chip for comparative proteomic analysis of isotopically labeled MCF-7 breast cancer cell extracts is explored for the first time.

  13. Droplet Microfluidics for Chip-Based Diagnostics

    PubMed Central

    Kaler, Karan V. I. S.; Prakash, Ravi

    2014-01-01

    Droplet microfluidics (DMF) is a fluidic handling technology that enables precision control over dispensing and subsequent manipulation of droplets in the volume range of microliters to picoliters, on a micro-fabricated device. There are several different droplet actuation methods, all of which can generate external stimuli, to either actively or passively control the shape and positioning of fluidic droplets over patterned substrates. In this review article, we focus on the operation and utility of electro-actuation-based DMF devices, which utilize one or more micro-/nano-patterned substrates to facilitate electric field-based handling of chemical and/or biological samples. The underlying theory of DMF actuations, device fabrication methods and integration of optical and opto-electronic detectors is discussed in this review. Example applications of such electro-actuation-based DMF devices have also been included, illustrating the various actuation methods and their utility in conducting chip-based laboratory and clinical diagnostic assays. PMID:25490590

  14. Integrated polymerase chain reaction chips utilizing digital microfluidics.

    PubMed

    Chang, Yi-Hsien; Lee, Gwo-Bin; Huang, Fu-Chun; Chen, Yi-Yu; Lin, Jr-Lung

    2006-09-01

    This study reports an integrated microfluidic chip for polymerase chain reaction (PCR) applications utilizing digital microfluidic chip (DMC) technology. Several crucial procedures including sample transportation, mixing, and DNA amplification were performed on the integrated chip using electro-wetting-on-dielectric (EWOD) effect. An innovative concept of hydrophobic/hydrophilic structure has been successfully demonstrated to integrate the DMC chip with the on-chip PCR device. Sample droplets were generated, transported and mixed by the EWOD-actuation. Then the mixture droplets were transported to a PCR chamber by utilizing the hydrophilic/hydrophobic interface to generate required surface tension gradient. A micro temperature sensor and two micro heaters inside the PCR chamber along with a controller were used to form a micro temperature control module, which could perform precise PCR thermal cycling for DNA amplification. In order to demonstrate the performance of the integrated DMC/PCR chips, a detection gene for Dengue II virus was successfully amplified and detected. The new integrated DMC/PCR chips only required an operation voltage of 12V(RMS) at a frequency of 3 KHz for digital microfluidic actuation and 9V(DC) for thermal cycling. When compared to its large-scale counterparts for DNA amplification, the developed system consumed less sample and reagent and could reduce the detection time. The developed chips successfully demonstrated the feasibility of Lab-On-a-Chip (LOC) by utilizing EWOD-based digital microfluidics.

  15. USB-driven microfluidic chips on printed circuit boards.

    PubMed

    Li, Jiang; Wang, Yixuan; Dong, Enkai; Chen, Haosheng

    2014-03-07

    A technology is presented to fabricate a microfluidic chip in which the microchannels and the microelectrodes of sensors are integrated directly into the copper sheet on a printed circuit board. Then, we demonstrate an application of the generation of oil-in-water and water-in-oil emulsion droplets on this microfluidic chip driven by a USB interface, and the droplet size is detected by the microelectrodes on the downstream microchannel. The integration of the microfluidic chip is improved by the direct connection of the channels to the microelectrodes of the driving unit and of the sensors on the same substrate, and it is a promising way to integrate microfluidics into a more complex micro electrical-mechanical system (MEMS).

  16. Directed evolution of enzymes using microfluidic chips

    NASA Astrophysics Data System (ADS)

    Pilát, Zdeněk.; Ježek, Jan; Šmatlo, Filip; Kaůka, Jan; Zemánek, Pavel

    2016-12-01

    Enzymes are highly versatile and ubiquitous biological catalysts. They can greatly accelerate large variety of reactions, while ensuring appropriate catalytic activity and high selectivity. These properties make enzymes attractive biocatalysts for a wide range of industrial and biomedical applications. Over the last two decades, directed evolution of enzymes has transformed the field of protein engineering. We have devised microfluidic systems for directed evolution of haloalkane dehalogenases in emulsion droplets. In such a device, individual bacterial cells producing mutated variants of the same enzyme are encapsulated in microdroplets and supplied with a substrate. The conversion of a substrate by the enzyme produced by a single bacterium changes the pH in the droplet which is signalized by pH dependent fluorescence probe. The droplets with the highest enzymatic activity can be separated directly on the chip by dielectrophoresis and the resultant cell lineage can be used for enzyme production or for further rounds of directed evolution. This platform is applicable for fast screening of large libraries in directed evolution experiments requiring mutagenesis at multiple sites of a protein structure.

  17. Microfluidic cell chips for high-throughput drug screening.

    PubMed

    Chi, Chun-Wei; Ahmed, Ah Rezwanuddin; Dereli-Korkut, Zeynep; Wang, Sihong

    2016-05-01

    The current state of screening methods for drug discovery is still riddled with several inefficiencies. Although some widely used high-throughput screening platforms may enhance the drug screening process, their cost and oversimplification of cell-drug interactions pose a translational difficulty. Microfluidic cell-chips resolve many issues found in conventional HTS technology, providing benefits such as reduced sample quantity and integration of 3D cell culture physically more representative of the physiological/pathological microenvironment. In this review, we introduce the advantages of microfluidic devices in drug screening, and outline the critical factors which influence device design, highlighting recent innovations and advances in the field including a summary of commercialization efforts on microfluidic cell chips. Future perspectives of microfluidic cell devices are also provided based on considerations of present technological limitations and translational barriers.

  18. Integrated microfluidic linking chip for scanning probe nanolithography

    NASA Astrophysics Data System (ADS)

    Ryu, Kee Suk; Wang, Xuefeng; Shaikh, Kashan; Bullen, David; Goluch, Edgar; Zou, Jun; Liu, Chang; Mirkin, Chad A.

    2004-07-01

    This letter reports an architecture for a microfluidic chip that dresses (inks) multiple nanolithography tips in a high-density array in a parallel and multiplexed fashion. The microfluidic chip consists of multiple precision patterned thin-film poly(dimethylsiloxane) (PDMS) patches serving as porous inking pads. Inking chemicals are supplied from loading reservoirs to the inking pads through microfluidic channels. The gas-permeable thin PDMS membranes allow ink molecules to diffuse through while preventing bulk liquid from overflowing or evaporating. The inking chip provides high-density inking, easy loading of inks, and reduced evaporation losses. We present the fabrication process and inking of scanning probe contact printing probes and commercial nitride probes.

  19. Various On-Chip Sensors with Microfluidics for Biological Applications

    PubMed Central

    Lee, Hun; Xu, Linfeng; Koh, Domin; Nyayapathi, Nikhila; Oh, Kwang W.

    2014-01-01

    In this paper, we review recent advances in on-chip sensors integrated with microfluidics for biological applications. Since the 1990s, much research has concentrated on developing a sensing system using optical phenomena such as surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS) to improve the sensitivity of the device. The sensing performance can be significantly enhanced with the use of microfluidic chips to provide effective liquid manipulation and greater flexibility. We describe an optical image sensor with a simpler platform for better performance over a larger field of view (FOV) and greater depth of field (DOF). As a new trend, we review consumer electronics such as smart phones, tablets, Google glasses, etc. which are being incorporated in point-of-care (POC) testing systems. In addition, we discuss in detail the current optical sensing system integrated with a microfluidic chip. PMID:25222033

  20. On-chip Microfluidic Multimodal Swimmer toward 3D Navigation

    NASA Astrophysics Data System (ADS)

    Barbot, Antoine; Decanini, Dominique; Hwang, Gilgueng

    2016-01-01

    Mobile microrobots have a promising future in various applications. These include targeted drug delivery, local measurement, biopsy or microassembly. Studying mobile microrobots inside microfluidics is an essential step towards such applications. But in this environment that was not designed for the robot, integration process and propulsion robustness still pose technological challenges. In this paper, we present a helical microrobot with three different motions, designed to achieve these goals. These motions are rolling, spintop motion and swimming. Through these multiple motions, microrobots are able to selectively integrate a chip through a microfluidic channel. This enables them to perform propulsion characterizations, 3D (Three Dimensional) maneuverability, particle cargo transport manipulation and exit from the chip. The microrobot selective integration inside microfluidics could lead to various in-vitro biologic or in-vivo biomedical applications.

  1. On-chip Microfluidic Multimodal Swimmer toward 3D Navigation

    PubMed Central

    Barbot, Antoine; Decanini, Dominique; Hwang, Gilgueng

    2016-01-01

    Mobile microrobots have a promising future in various applications. These include targeted drug delivery, local measurement, biopsy or microassembly. Studying mobile microrobots inside microfluidics is an essential step towards such applications. But in this environment that was not designed for the robot, integration process and propulsion robustness still pose technological challenges. In this paper, we present a helical microrobot with three different motions, designed to achieve these goals. These motions are rolling, spintop motion and swimming. Through these multiple motions, microrobots are able to selectively integrate a chip through a microfluidic channel. This enables them to perform propulsion characterizations, 3D (Three Dimensional) maneuverability, particle cargo transport manipulation and exit from the chip. The microrobot selective integration inside microfluidics could lead to various in-vitro biologic or in-vivo biomedical applications. PMID:26791433

  2. A microfluidic microprocessor: controlling biomimetic containers and cells using hybrid integrated circuit/microfluidic chips.

    PubMed

    Issadore, David; Franke, Thomas; Brown, Keith A; Westervelt, Robert M

    2010-11-07

    We present an integrated platform for performing biological and chemical experiments on a chip based on standard CMOS technology. We have developed a hybrid integrated circuit (IC)/microfluidic chip that can simultaneously control thousands of living cells and pL volumes of fluid, enabling a wide variety of chemical and biological tasks. Taking inspiration from cellular biology, phospholipid bilayer vesicles are used as robust picolitre containers for reagents on the chip. The hybrid chip can be programmed to trap, move, and porate individual living cells and vesicles and fuse and deform vesicles using electric fields. The IC spatially patterns electric fields in a microfluidic chamber using 128 × 256 (32,768) 11 × 11 μm(2) metal pixels, each of which can be individually driven with a radio frequency (RF) voltage. The chip's basic functions can be combined in series to perform complex biological and chemical tasks and can be performed in parallel on the chip's many pixels for high-throughput operations. The hybrid chip operates in two distinct modes, defined by the frequency of the RF voltage applied to the pixels: Voltages at MHz frequencies are used to trap, move, and deform objects using dielectrophoresis and voltages at frequencies below 1 kHz are used for electroporation and electrofusion. This work represents an important step towards miniaturizing the complex chemical and biological experiments used for diagnostics and research onto automated and inexpensive chips.

  3. Polymer microfluidic chip for online monitoring of microarray hybridizations.

    PubMed

    Noerholm, Mikkel; Bruus, Henrik; Jakobsen, Mogens H; Telleman, Pieter; Ramsing, Niels B

    2004-02-01

    A disposable single use polymer microfluidics chip has been developed and manufactured by micro injection molding. The chip has the same outer dimensions as a standard microscope slide (25 x 76 x 1.1 mm) and is designed to be compatible with existing microscope slide handling equipment like microarray scanners. The chip contains an inlet, a 10 microL hybridization chamber capable of holding a 1000 spot array, a waste chamber and a vent to allow air to escape when sample is injected. The hybridization chamber ensures highly homogeneous hybridization conditions across the microarray. We describe the use of this chip in a flexible setup with fluorescence based detection, temperature control and liquid handling by computer controlled syringe pumps. The chip and the setup presented in this article provide a powerful tool for highly parallel studies of kinetics and thermodynamics of duplex formation in DNA microarrays. The experimental setup presented in this article enables the on-chip microarray to be hybridized and monitored at several different stringency conditions during a single assay. The performance of the chip and the setup is demonstrated by on-line measurements of a hybridization of a DNA target solution to a microarray. A presented numerical model indicates that the hybridization process in microfluidic hybridization assays is diffusion limited, due to the low values of the diffusion coefficients D of the DNA and RNA molecules involved.

  4. Detection and classification of ebola on microfluidic chips

    NASA Astrophysics Data System (ADS)

    Lin, Xue; Jin, Xiangyu; Fan, Yunqian; Huang, Qin; Kou, Yue; Zu, Guo; Huang, Shiguang; Liu, Xiaosheng; Huang, Guoliang

    2016-10-01

    Point-of-care testing (POCT) for an infectious diseases is the prerequisite to control of the disease and limitation of its spread. A microfluidic chip for detection and classification of four strains of Ebola virus was developed and evaluated. This assay was based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific primers for Ebola Zaire virus, Ebola Sudan virus, Ebola Tai Forest virus and Ebola Bundibugyo virus were designed. The sensitivity of the microfluidic chip was under 103 copies per milliliter, as determined by ten repeated tests. This assay is unique in its ability to enable diagnosis of the Ebola infections and simultaneous typing of Ebola virus on a single chip. It offers short reaction time, ease of use and high specificity. These features should enable POCT in remote area during outbreaks of Ebola virus.

  5. [Application of microfluidic-chip in biomedicine].

    PubMed

    Bi, Ying-Nan; Zhang, Hui-Jing

    2006-01-01

    As a novel analytical technology, the research of Micro total analysis systems (micro-TAS) has been spreading rapidly. micro-TAS has been widely used to perform chemical and biochemical analysis. Microfluidic-based analytical system as micro-TAS's manily direction develops very fast in terms of it's reaction speed, reagent consumption, miniaturization, cost, and automation. After having proven the value of microfluidics for genetic, proteomic and cytomics analysis, this article also anticipates the development tendency of this technology in the biology medicine domain. It has demonstrated that a truly, easy-to-handle Microfluidic-based analytical device will be emerged in the future.

  6. The microfluidic puzzle: chip-oriented rapid prototyping.

    PubMed

    Lim, Jiseok; Maes, Florine; Taly, Valérie; Baret, Jean-Christophe

    2014-05-21

    We demonstrate a new concept for reconfigurable microfluidic devices from elementary functional units. Our approach suppresses the need for patterning, soft molding and bonding when details on a chip have to be modified. Our system has two parts, a base-platform used as a scaffold and functional modules which are combined by 'plug-and-play'. To demonstrate that our system sustains typical pressures in microfluidic experiments, we produce droplets of different sizes using T-junction modules with three different designs assembled successively on a 3 × 3 modular scaffold.

  7. Seamless integration of CMOS and microfluidics using flip chip bonding

    NASA Astrophysics Data System (ADS)

    Welch, David; Blain Christen, Jennifer

    2013-03-01

    We demonstrate the microassembly of PDMS (polydimethylsiloxane) microfluidics with integrated circuits made in complementary metal-oxide-semiconductor (CMOS) processes. CMOS-sized chips are flip chip bonded to a flexible polyimide printed circuit board (PCB) with commercially available solder paste patterned using a SU-8 epoxy. The average resistance of each flip chip bond is negligible and all connections are electrically isolated. PDMS is attached to the flexible polyimide PCB using a combination of oxygen plasma treatment and chemical bonding with 3-aminopropyltriethoxysilane. The total device has a burst pressure of 175 kPA which is limited by the strength of the flip chip attachment. This technique allows the sensor area of the die to act as the bottom of the microfluidic channel. The SU-8 provides a barrier between the pad ring (electrical interface) and the fluids; post-processing is not required on the CMOS die. This assembly method shows great promise for developing analytic systems which combine the strengths of microelectronics and microfluidics into one device.

  8. The promise of macromolecular crystallization in microfluidic chips

    NASA Technical Reports Server (NTRS)

    van der Woerd, Mark; Ferree, Darren; Pusey, Marc

    2003-01-01

    Microfluidics, or lab-on-a-chip technology, is proving to be a powerful, rapid, and efficient approach to a wide variety of bioanalytical and microscale biopreparative needs. The low materials consumption, combined with the potential for packing a large number of experiments in a few cubic centimeters, makes it an attractive technique for both initial screening and subsequent optimization of macromolecular crystallization conditions. Screening operations, which require a macromolecule solution with a standard set of premixed solutions, are relatively straightforward and have been successfully demonstrated in a microfluidics platform. Optimization methods, in which crystallization solutions are independently formulated from a range of stock solutions, are considerably more complex and have yet to be demonstrated. To be competitive with either approach, a microfluidics system must offer ease of operation, be able to maintain a sealed environment over several weeks to months, and give ready access for the observation and harvesting of crystals as they are grown.

  9. High Voltage Dielectrophoretic and Magnetophoretic Hybrid Integrated Circuit / Microfluidic Chip.

    PubMed

    Issadore, David; Franke, Thomas; Brown, Keith A; Hunt, Thomas P; Westervelt, Robert M

    2009-12-01

    A hybrid integrated circuit (IC) / microfluidic chip is presented that independently and simultaneously traps and moves microscopic objects suspended in fluid using both electric and magnetic fields. This hybrid chip controls the location of dielectric objects, such as living cells and drops of fluid, on a 60 × 61 array of pixels that are 30 × 38 μm(2) in size, each of which can be individually addressed with a 50 V peak-to-peak, DC to 10 MHz radio frequency voltage. These high voltage pixels produce electric fields above the chip's surface with a magnitude , resulting in strong dielectrophoresis (DEP) forces . Underneath the array of DEP pixels there is a magnetic matrix that consists of two perpendicular sets of 60 metal wires running across the chip. Each wire can be sourced with 120 mA to trap and move magnetically susceptible objects using magnetophoresis (MP). The DEP pixel array and magnetic matrix can be used simultaneously to apply forces to microscopic objects, such as living cells or lipid vesicles, that are tagged with magnetic nanoparticles. The capabilities of the hybrid IC / microfluidic chip demonstrated in this paper provide important building blocks for a platform for biological and chemical applications.

  10. A glass microfluidic chip adhesive bonding method at room temperature

    NASA Astrophysics Data System (ADS)

    Pan, Yu-Jen; Yang, Ruey-Jen

    2006-12-01

    This paper presents a novel method using UV epoxy resin for the bonding of glass blanks and patterned plates at room temperature. There is no need to use a high-temperature thermal fusion process and therefore avoid damaging temperature-sensitive metals in a microchip. The proposed technique has the further advantage that the sealed glass blanks and patterned plates can be separated by the application of adequate heat. In this way, the microchip can be opened, the fouling microchannels may be easily cleaned-up and the plates then re-bonded to recycle the microchip. The proposed sealing method is used to bond a microfluidic device, and the bonding strength is then investigated in a series of chemical resistance tests conducted in various chemicals. Leakage of solution was evaluated in a microfluidic chip using pressure testing to 1.792 × 102 kPa (26 psi), and the microchannel had no observable leak. Electrical leakage between channels was tested by comparing the resistances of two bonding methods, and the result shows no significant electrical leakage. The performance of the device obtained from the proposed bonding method is compared with that of the thermal fusion bonding technique for an identical microfluidic device. It is found that identical results are obtained under the same operating conditions. The proposed method provides a simple, quick and inexpensive method for sealing glass microfluidic chips.

  11. Distillation and detection of SO2 using a microfluidic chip.

    PubMed

    Ju, Wei-Jhong; Fu, Lung-Ming; Yang, Ruey-Jen; Lee, Chia-Lun

    2012-02-07

    A miniaturized distillation system is presented for separating sulfurous acid (H(2)SO(3)) into sulfur dioxide (SO(2)) and water (H(2)O). The major components of the proposed system include a microfluidic distillation chip, a power control module, and a carrier gas pressure control module. The microfluidic chip is patterned using a commercial CO(2) laser and comprises a serpentine channel, a heating zone, a buffer zone, a cooling zone, and a collection tank. In the proposed device, the H(2)SO(3) solution is injected into the microfluidic chip and is separated into SO(2) and H(2)O via an appropriate control of the distillation time and temperature. The gaseous SO(2) is then transported into the collection chamber by the carrier gas and is mixed with DI water. Finally, the SO(2) concentration is deduced from the absorbance measurements obtained using a spectrophotometer. The experimental results show that a correlation coefficient of R(2) = 0.9981 and a distillation efficiency as high as 94.6% are obtained for H(2)SO(3) solutions with SO(2) concentrations in the range of 100-500 ppm. The SO(2) concentrations of two commercial red wines are successfully detected using the developed device. Overall, the results presented in this study show that the proposed system provides a compact and reliable tool for SO(2) concentration measurement purposes.

  12. Design and research for biosensing THz microfluidic chips

    NASA Astrophysics Data System (ADS)

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

    2016-11-01

    Many Biomolecules vibration frequencies are in terahertz (0.1THz-10THz) frequency range, so terahertz (THz) technology is an essential tool for detecting biological molecules. However, due to terahertz strongly absorbed by water, it is difficult to detect these molecules for biological and chemical liquid samples. Therefore, we present a novel detection method by combining terahertz technology with microfluidic technology. The strong absorption of water is effectively overcome by controlling the length that terahertz passes through liquid samples. What's more, a higher signal to noise ratio is obtained through using less samples. In this paper, we designed a THz microfluidic chip that is easy to be fabricated by using the materials of Zeonor and polydimethylsiloxane (PDMS). Using terahertz time-domainspectroscopy (THz-TDS) system, we find that the chip has a high transmittance above 80% in the range from 0.2 THz to 2.6 THz. Then the THz spectra of deionized water and different kinds of solutions with different concentrations in the microfluidic chip were measured, respectively. In our research, it is found that different kinds of solutions have different transmission coefficients for THz. In addition, the THz transmission and absorption spectrum changes with the concentration of the same kind of solution.

  13. Structural Characterization of a Capillary Microfluidic Chip Using Microreflectance.

    PubMed

    Lastras-Martínez, Luis F; Balderas-Navarro, Raul E; Castro-García, Ricardo; Hernández-Vidales, Karen; Almendarez-Rodríguez, Juan; Herrera-Jasso, Rafael; Prinz, Adrian; Bergmair, Iris

    2016-10-18

    The structural characterization of capillary microfluidic chips is important for reliable applications. In particular, nondestructive diagnostic tools to assess geometrical dimensions and their correlations with control processes are of much importance, preferably if they are implemented in situ. Several techniques to accomplish this task have been reported; namely, optical coherence tomography (OCT) jointly with confocal fluorescence microscopy (CFM) to investigate internal features of lab-on-a-chip technologies. In this paper, we report on the use of a simple optical technique, based on near-normal incidence microreflectance, which allows mapping internal features of a microfluidic chip in a straightforward way. Our setup is based on a charge-coupled device camera that allows a lateral resolution of ∼2.5 µm and allows us to measure in the wavelength range of 640-750 nm. The technique takes advantage of the Fabry-Perot interferences features in the reflectance spectra, which are further analyzed by a discrete Fourier transform. In this way, the amplitude of the Fourier coefficients is modulated by the presence of a microfluidic channel.

  14. Note: A microfluidic chip setup for capillarity-assisted particle assembly

    NASA Astrophysics Data System (ADS)

    Klein, M. J. K.; Kuemin, C.; Tamulevicius, T.; Manning, M.; Wolf, H.

    2012-08-01

    We developed a microfluidic chip setup for capillarity-assisted particle assembly (CAPA). A capillary bridge is formed between the aperture of a silicon chip and the assembly template. The bridge is fed with particle suspension through a microfluidic channel on the chip top side. With this setup, we can control the particle assembly location and tune the suspension composition during particle assembly. In this note, we describe the chip setup, the CAPA process using the microfluidic chip, and results of complex particle assemblies, such as composite particle arrays and particle gradients, that could not be obtained using a conventional CAPA setup.

  15. High Voltage Dielectrophoretic and Magnetophoretic Hybrid Integrated Circuit / Microfluidic Chip

    PubMed Central

    Issadore, David; Franke, Thomas; Brown, Keith A.; Hunt, Thomas P.; Westervelt, Robert M.

    2010-01-01

    A hybrid integrated circuit (IC) / microfluidic chip is presented that independently and simultaneously traps and moves microscopic objects suspended in fluid using both electric and magnetic fields. This hybrid chip controls the location of dielectric objects, such as living cells and drops of fluid, on a 60 × 61 array of pixels that are 30 × 38 μm2 in size, each of which can be individually addressed with a 50 V peak-to-peak, DC to 10 MHz radio frequency voltage. These high voltage pixels produce electric fields above the chip’s surface with a magnitude , resulting in strong dielectrophoresis (DEP) forces . Underneath the array of DEP pixels there is a magnetic matrix that consists of two perpendicular sets of 60 metal wires running across the chip. Each wire can be sourced with 120 mA to trap and move magnetically susceptible objects using magnetophoresis (MP). The DEP pixel array and magnetic matrix can be used simultaneously to apply forces to microscopic objects, such as living cells or lipid vesicles, that are tagged with magnetic nanoparticles. The capabilities of the hybrid IC / microfluidic chip demonstrated in this paper provide important building blocks for a platform for biological and chemical applications. PMID:20625468

  16. Microfluidic-Based sample chips for radioactive solutions

    SciTech Connect

    Tripp, J. L.; Law, J. D.; Smith, T. E.; Rutledge, V. J.; Bauer, W. F.; Ball, R. D.; Hahn, P. A.

    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 would 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.

  17. Microfluidic-Based Sample Chips for Radioactive Solutions

    SciTech Connect

    Tripp, J. L.; Law, J. D.; Smith, T. E.; Rutledge, V. J.; Bauer, W. F.; Ball, R. D.; Hahn, P. A.

    2014-02-01

    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 would 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.

  18. Microfluidic Chip Coupled with Thermal Desorption Atmospheric Pressure Ionization Mass Spectrometry

    PubMed Central

    Chang, Chia-Hsien; Chen, Tsung-Yi; Chen, Yu-Chie

    2014-01-01

    Microfluidic chips have been used as platforms for a diversity of research purposes such as for separation and micro-reaction. One of the suitable detectors for microfluidic chip is mass spectrometry. Because microfluidic chips are generally operated in an open air condition, mass spectrometry coupled with atmospheric pressure ion sources can suit the requirement with minimum compromise. In this study, we develop a new interface to couple a microfluidic chip with mass spectrometry. A capillary tip coated with a layer of graphite, capable of absorbing energy of near-infrared (NIR) light is used to interface microfluidic chip with mass spectrometry. An NIR laser diode (λ=808 nm) is used to irradiate the capillary tip for assisting the generation of spray from the eluent of the microfluidic chip. An electrospray is provided to fuse with the spray generated from the microfluidic chip for post-ionization. Transesterification is used as the example to demonstrate the feasibility of using this interface to couple microfluidic chip with mass spectrometry. PMID:26839753

  19. Polydimethylsiloxane-based conducting composites and their applications in microfluidic chip fabrication

    PubMed Central

    Gong, Xiuqing; Wen, Weijia

    2009-01-01

    This paper reviews the design and fabrication of polydimethylsiloxane (PDMS)-based conducting composites and their applications in microfluidic chip fabrication. Owing to their good electrical conductivity and rubberlike elastic characteristics, these composites can be used variously in soft-touch electronic packaging, planar and three-dimensional electronic circuits, and in-chip electrodes. Several microfluidic components fabricated with PDMS-based composites have been introduced, including a microfluidic mixer, a microheater, a micropump, a microdroplet controller, as well as an all-in-one microfluidic chip. PMID:19693388

  20. A novel monolithic fabrication method for a plastic microfluidic chip with liquid interconnecting ports

    NASA Astrophysics Data System (ADS)

    Lee, Bong-Kee; Kwon, Tai Hun

    2010-10-01

    In the present study, a novel monolithic fabrication method was developed for the manufacturing of a plastic microfluidic chip with liquid interconnecting ports. As the present method can realize both interconnecting ports and through holes, which are essential components for the delivery of working fluids, in the plastic microfluidic chip, no additional processes and external ports are required. Furthermore, the connection of external silicone tubing can be simply achieved by utilizing an elastic deformation of the used tubing. As one representative example, a microinjection molding of a prototype microfluidic chip having two types of interconnecting ports was demonstrated. After obtaining upper and lower plates by the microinjection molding process utilizing mold cores with pin structures, the thermal bonding of the molded plates was carried out, resulting in the prototype plastic microfluidic chip with interconnecting ports. From microfluidic experiments using the fabricated prototype, it was found that the present method could be quite useful in various microfluidic applications.

  1. Fabrication and multifunction integration of microfluidic chips by femtosecond laser direct writing.

    PubMed

    Xu, Bin-Bin; Zhang, Yong-Lai; Xia, Hong; Dong, Wen-Fei; Ding, Hong; Sun, Hong-Bo

    2013-05-07

    In the pursuit of modern microfluidic chips with multifunction integration, micronanofabrication techniques play an increasingly important role. Despite the fact that conventional fabrication approaches such as lithography, imprinting and soft lithography have been widely used for the preparation of microfluidic chips, it is still challenging to achieve complex microfluidic chips with multifunction integration. Therefore, novel micronanofabrication approaches that could be used to achieve this end are highly desired. As a powerful 3D processing tool, femtosecond laser fabrication shows great potential to endow general microfluidic chips with multifunctional units. In this review, we briefly introduce the fundamental principles of femtosecond laser micronanofabrication. With the help of laser techniques, both the preparation and functionalization of advanced microfluidic chips are summarized. Finally, the current challenges and future perspective of this dynamic field are discussed based on our own opinion.

  2. Microfluidic-chip platform for cell sorting

    NASA Astrophysics Data System (ADS)

    Malik, Sarul; Balyan, Prerna; Akhtar, J.; Agarwal, Ajay

    2016-04-01

    Cell sorting and separation are considered to be very crucial preparatory steps for numerous clinical diagnostics and therapeutics applications in cell biology research arena. Label free cell separation techniques acceptance rate has been increased to multifold by various research groups. Size based cell separation method focuses on the intrinsic properties of the cell which not only avoids clogging issues associated with mechanical and centrifugation filtration methods but also reduces the overall cost for the process. Consequentially flow based cell separation method for continuous flow has attracted the attention of millions. Due to the realization of structures close to particle size in micro dimensions, the microfluidic devices offer precise and rapid particle manipulation which ultimately leads to an extraordinary cell separation results. The proposed microfluidic device is fabricated to separate polystyrene beads of size 1 µm, 5 µm, 10 µm and 20 µm. The actual dimensions of blood corpuscles were kept in mind while deciding the particle size of polystyrene beads which are used as a model particles for study.

  3. The RootChip: an integrated microfluidic chip for plant science.

    PubMed

    Grossmann, Guido; Guo, Woei-Jiun; Ehrhardt, David W; Frommer, Wolf B; Sit, Rene V; Quake, Stephen R; Meier, Matthias

    2011-12-01

    Studying development and physiology of growing roots is challenging due to limitations regarding cellular and subcellular analysis under controlled environmental conditions. We describe a microfluidic chip platform, called RootChip, that integrates live-cell imaging of growth and metabolism of Arabidopsis thaliana roots with rapid modulation of environmental conditions. The RootChip has separate chambers for individual regulation of the microenvironment of multiple roots from multiple seedlings in parallel. We demonstrate the utility of The RootChip by monitoring time-resolved growth and cytosolic sugar levels at subcellular resolution in plants by a genetically encoded fluorescence sensor for glucose and galactose. The RootChip can be modified for use with roots from other plant species by adapting the chamber geometry and facilitates the systematic analysis of root growth and metabolism from multiple seedlings, paving the way for large-scale phenotyping of root metabolism and signaling.

  4. Microfluidics on liquid handling stations (μF-on-LHS): an industry compatible chip interface between microfluidics and automated liquid handling stations.

    PubMed

    Waldbaur, Ansgar; Kittelmann, Jörg; Radtke, Carsten P; Hubbuch, Jürgen; Rapp, Bastian E

    2013-06-21

    We describe a generic microfluidic interface design that allows the connection of microfluidic chips to established industrial liquid handling stations (LHS). A molding tool has been designed that allows fabrication of low-cost disposable polydimethylsiloxane (PDMS) chips with interfaces that provide convenient and reversible connection of the microfluidic chip to industrial LHS. The concept allows complete freedom of design for the microfluidic chip itself. In this setup all peripheral fluidic components (such as valves and pumps) usually required for microfluidic experiments are provided by the LHS. Experiments (including readout) can be carried out fully automated using the hardware and software provided by LHS manufacturer. Our approach uses a chip interface that is compatible with widely used and industrially established LHS which is a significant advancement towards near-industrial experimental design in microfluidics and will greatly facilitate the acceptance and translation of microfluidics technology in industry.

  5. A microwave resonator integrated on a polymer microfluidic chip.

    PubMed

    Kiss, S Z; Rostas, A M; Heidinger, L; Spengler, N; Meissner, M V; MacKinnon, N; Schleicher, E; Weber, S; Korvink, J G

    2016-09-01

    We describe a novel stacked split-ring type microwave (MW) resonator that is integrated into a 10mm by 10mm sized microfluidic chip. A straightforward and scalable batch fabrication process renders the chip suitable for single-use applications. The resonator volume can be conveniently loaded with liquid sample via microfluidic channels patterned into the mid layer of the chip. The proposed MW resonator offers an alternative solution for compact in-field measurements, such as low-field magnetic resonance (MR) experiments requiring convenient sample exchange. A microstrip line was used to inductively couple MWs into the resonator. We characterised the proposed resonator topology by electromagnetic (EM) field simulations, a field perturbation method, as well as by return loss measurements. Electron paramagnetic resonance (EPR) spectra at X-band frequencies were recorded, revealing an electron-spin sensitivity of 3.7·10(11)spins·Hz(-1/2)G(-1) for a single EPR transition. Preliminary time-resolved EPR experiments on light-induced triplet states in pentacene were performed to estimate the MW conversion efficiency of the resonator.

  6. A microwave resonator integrated on a polymer microfluidic chip

    NASA Astrophysics Data System (ADS)

    Kiss, S. Z.; Rostas, A. M.; Heidinger, L.; Spengler, N.; Meissner, M. V.; MacKinnon, N.; Schleicher, E.; Weber, S.; Korvink, J. G.

    2016-09-01

    We describe a novel stacked split-ring type microwave (MW) resonator that is integrated into a 10 mm by 10 mm sized microfluidic chip. A straightforward and scalable batch fabrication process renders the chip suitable for single-use applications. The resonator volume can be conveniently loaded with liquid sample via microfluidic channels patterned into the mid layer of the chip. The proposed MW resonator offers an alternative solution for compact in-field measurements, such as low-field magnetic resonance (MR) experiments requiring convenient sample exchange. A microstrip line was used to inductively couple MWs into the resonator. We characterised the proposed resonator topology by electromagnetic (EM) field simulations, a field perturbation method, as well as by return loss measurements. Electron paramagnetic resonance (EPR) spectra at X-band frequencies were recorded, revealing an electron-spin sensitivity of 3.7 ·1011spins ·Hz - 1 / 2G-1 for a single EPR transition. Preliminary time-resolved EPR experiments on light-induced triplet states in pentacene were performed to estimate the MW conversion efficiency of the resonator.

  7. Microfluidics without channels: highly-flexible synthesis on a digital-microfluidic chip for production of diverse PET tracers

    SciTech Connect

    Van Dam, Robert Michael

    2010-09-01

    Positron emission tomography (PET) imaging is used for fundamental studies of living biological organisms and microbial ecosystems in applications ranging from biofuel production to environmental remediation to the study, diagnosis, and treatment monitoring of human disease. Routine access to PET imaging, to monitor biochemical reactions in living organisms in real time, could accelerate a broad range of research programs of interest to DOE. Using PET requires access to short-lived radioactive-labeled compounds that specifically probe the desired living processes. The overall aims of this project were to develop a miniature liquid-handling technology platform (called “microfluidics”) that increases the availability of diverse PET probes by reducing the cost and complexity of their production. Based on preliminary experiments showing that microfluidic chips can synthesis such compounds, we aimed to advance this technology to improve its robustness, increase its flexibility for a broad range of probes, and increase its user-friendliness. Through the research activities of this project, numerous advances were made; Tools were developed to enable the visualization of radioactive materials within microfluidic chips; Fundamental advances were made in the microfluidic chip architecture and fabrication process to increase its robustness and reliability; The microfluidic chip technology was shown to produce useful quantities of an example PET probes, and methods to further increase the output were successfully pursued; A “universal” chip was developed that could produce multiple types of PET probes, enabling the possibility of “on demand” synthesis of different probes; and Operation of the chip was automated to ensure minimal radiation exposure to the operator Based on the demonstrations of promising technical feasibility and performance, the microfluidic chip technology is currently being commercialized. It is anticipated that costs of microfluidic chips can be

  8. 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.

  9. Microfluidic cytometers with integrated on-chip optical systems for red blood cell and platelet counting.

    PubMed

    Zhao, Yingying; Li, Qin; Hu, Xiaoming; Lo, Yuhwa

    2016-11-01

    A microfluidic cytometer with integrated on-chip optical systems was designed for red blood cell (RBC) and platelet (PLT) counting. The design, fabrication, and characterization of the microfluidic cytometer with on-chip optical signal detection were described. With process using only a single mask, the device that integrates optical fibers and on-chip microlens with microfluidic channels on a polydimethylsiloxane layer by standard soft photolithography. This compact structure increased the sensitivity of the device and eliminated time-consuming free-space optical alignments. The microfluidic cytometer was used to count red blood cells and platelets. Forward scatter and extinction were collected simultaneously for each cell. Experimental results indicated that the microfluidic cytometer exhibited comparable performance with a conventional cytometer and demonstrated superior capacity to detect on-chip optical signals in a highly compact, simple, truly portable, and low-cost format that is well suitable for point-of-care clinical diagnostics.

  10. ITP of lanthanides in microfluidic PMMA chip.

    PubMed

    Cong, Yongzheng; Bottenus, Danny; Liu, Bingwen; Clark, Sue B; Ivory, Cornelius F

    2014-03-01

    An ITP separation of eight lanthanides on a serpentine PMMA microchip with a tee junction and a 230-mm-long serpentine channel is described. The cover of the PMMA chip is 175 μm thick so that a C(4) D in microchip mode can be used to detect the lanthanides as they migrate through the microchannel. Acetate and α-hydroxyisobutyric acid are used as complexing agents to increase the electrophoretic mobility difference between the lanthanides. Eight lanthanides are concentrated within ∼ 6 min by ITP in the microchip using 10 mM ammonium acetate at pH 4.5 as the leading electrolyte and 10 mM acetic acid at ∼ pH 3.0 as the terminating electrolyte. In addition, a 2D numerical simulation of the lanthanides undergoing ITP in the microchip is compared with experimental results using COMSOL Multiphysics v4.3a.

  11. Microfluidic chips with reversed-phase monoliths for solid phase extraction and on-chip labeling.

    PubMed

    Nge, Pamela N; Pagaduan, Jayson V; Yu, Ming; Woolley, Adam T

    2012-10-26

    The integration of sample preparation methods into microfluidic devices provides automation necessary for achieving complete micro total analysis systems. We have developed a technique that combines on-chip sample enrichment with fluorescence labeling and purification. Polymer monoliths made from butyl methacrylate were fabricated in cyclic olefin copolymer microdevices and used for solid phase extraction. We studied the retention of fluorophores, amino acids and proteins on these columns. The retained samples were subsequently labeled with both Alexa Fluor 488 and Chromeo P503, and unreacted dye was rinsed off the column before sample elution. Additional purification was obtained from the differential retention of proteins and fluorescent labels. A linear relation between the eluted peak areas and concentrations of on-chip labeled heat shock protein 90 samples demonstrated the utility of this method for on-chip quantitation. Our fast and simple method of simultaneously concentrating and labeling samples on-chip is compatible with miniaturization and desirable for automated analysis.

  12. Polymeric nanofiber web-based artificial renal microfluidic chip.

    PubMed

    Lee, K H; Kim, D J; Min, B G; Lee, S H

    2007-08-01

    In this paper, we present a new method for the creation of a smaller dialyzer and do so by incorporating polymeric nanofiber web, which is known to have good filtration efficiency for broad particle sizes, into a poly (dimethylsiloxane)-based microplatform. We have developed a process that makes possible the efficient production of polyethersulfone and polyurethane nanofiber web and that, itself, incorporates an electrospinning method. We have combined the nanofiber web with the PDMS-based microfluidic platform to create a chip-based portable hemodialysis system. With the dialyzing chip, we evaluated the filtration capability of molecules in broad ranges of sizes and compared the filtration capability of nanofiber membranes with that of PES and polyvinylidene fluoride porous membranes (sheet type): we discovered that the nanofiber membranes have better filtration performance than the other membranes. Blood cells were not mechanically affected during their filtration and their transportation through the chip. In conclusion, we have demonstrated the feasibility of chip-based hemodialysis, and we expect that our method suggested in this paper will be applied to the development of small light-weight dialyzers for the realization of portable hemodialysis systems.

  13. Development of a microfluidic chip-based plasmid miniprep.

    PubMed

    Northrup, Victoria A; Backhouse, Christopher J; Glerum, D Moira

    2010-07-15

    Plasmids are the workhorse of contemporary molecular biology, serving as vectors in the multitude of molecular cloning approaches now available. Plasmid minipreps are a routine and essential means of extracting plasmid DNA from bacteria, such as Escherichia coli, for identification, characterization, and further manipulation. Although there have been many approaches described and miniprep kits are commercially available, traditional minipreps typically require more than 16h, including the time needed for bacterial cell culture. Here we describe the development of a microfluidic chip (MFC)-based miniprep that uses on-chip lysis and trapping of large DNA in agarose to differentially separate plasmid DNA from the bacterial chromosome. Our approach greatly decreases both the time required for the miniprep itself and the time required for growth of the bacterial cultures because our on-chip miniprep uses 10(5) times fewer E. coli cells. Because the quality of the isolated plasmid is comparable to that obtained using conventional miniprep protocols, this approach allows growth of E. coli and isolation of plasmid within hours, thereby making it ideal for rapid screening approaches. This MFC-based miniprep, coupled with recently demonstrated on-chip transfection capabilities, lays the groundwork for seamless manipulation of plasmids on MFC platforms.

  14. The Promise of Macromolecular Crystallization in Micro-fluidic Chips

    NASA Technical Reports Server (NTRS)

    vanderWoerd, Mark; Ferree, Darren; Pusey, Marc

    2003-01-01

    Micro-fluidics, or lab on a chip technology, is proving to be a powerful, rapid, and efficient approach to a wide variety of bio-analytical and microscale bio-preparative needs. The low materials consumption, combined with the potential for packing a large number of experiments in a few cubic centimeters, makes it an attractive technique for both initial screening and subsequent optimization of macromolecular crystallization conditions. Screening operations, which require equilibrating macromolecule solution with a standard set of premixed solutions, are relatively straightforward and have been successfully demonstrated in a micro-fluidics platform. More complex optimization methods, where crystallization solutions are independently formulated from a range of stock solutions, are considerably more complex and have yet to be demonstrated. To be competitive with either approach, a micro-fluidics system must offer ease of operation, be able to maintain a sealed environment over several weeks to months, and give ready access for the observation of crystals as they are grown.

  15. Calibration of optical coherence tomography angiography with a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Su, Johnny P.; Chandwani, Rahul; Gao, Simon S.; Pechauer, Alex D.; Zhang, Miao; Wang, Jie; Jia, Yali; Huang, David; Liu, Gangjun

    2016-08-01

    A microfluidic chip with microchannels ranging from 8 to 96 μm was used to mimic blood vessels down to the capillary level. Blood flow within the microfluidic channels was analyzed with split-spectrum amplitude-decorrelation angiography (SSADA)-based optical coherence tomography (OCT) angiography. It was found that the SSADA decorrelation value was related to both blood flow speed and channel width. SSADA could differentiate nonflowing blood inside the microfluidic channels from static paper. The SSADA decorrelation value was approximately linear with blood flow velocity up to a threshold Vsat of 5.83±1.33 mm/s (mean±standard deviation over the range of channel widths). Beyond this threshold, it approached a saturation value Dsat. Dsat was higher for wider channels, and approached a maximum value Dsm as the channel width became much larger than the beam focal spot diameter. These results indicate that decorrelation values (flow signal) in capillary networks would be proportional to both flow velocity and vessel caliber but would be capped at a saturation value in larger blood vessels. These findings are useful for interpretation and quantification of clinical OCT angiography results.

  16. Optical two-beam trap in a polymer microfluidic chip

    NASA Astrophysics Data System (ADS)

    Espina Palanco, Marta; Catak, Darmin; Marie, Rodolphe; Matteucci, Marco; Bilenberg, Brian; Kristensen, Anders; Berg-Sørensen, Kirstine

    2016-09-01

    An optical two-beam trap, composed from two counter propagating laser beams, is an interesting setup due to the ability of the system to trap, hold, and stretch soft biological objects like vesicles or single cells. Because of this functionality, the system was also named "the optical stretcher" by Jochen Guck, Josep Käs and co-workers some 15 years ago. In a favorable setup, the two opposing laser beams meet with equal intensities in the middle of a fluidic channel in which cells may flow past, be trapped, stretched, and allowed to move on, giving the promise of a high throughput device. Yet, single beam optical traps, aka optical tweezers, by far outnumber the existing optical stretchers in research labs throughout the world. The ability to easily construct an optical stretcher setup in a low-cost material would possibly imply more frequent use of the optical stretching technique. Here, we will outline the design, the production procedures, and results obtained in a fiber-based experimental setup built within an injection molded microfluidic polymer chip. The microfluidic chip is constructed with a three layer technology in which we ensure both horizontal and vertical focusing of the cells we wish to trap, thereby preventing too many cells to flow below the line of focus of the two counter propagating laser beams that are positioned perpendicular to the direction of flow of the cells. Results will be compared to that from other designs from previous work in the group.

  17. Polymethylhydrosiloxane (PMHS) as a functional material for microfluidic chips

    NASA Astrophysics Data System (ADS)

    Lee, S. J.; Goedert, M.; Matyska, M. T.; Ghandehari, E. M.; Vijay, M.; Pesek, J. J.

    2008-02-01

    Polymethylhydrosiloxane (PMHS) has been investigated as a candidate material for microfluidic chips. The ability to modify the surface of PMHS by hydrosilation is particularly advantageous for separation processes. The chemical modification of PMHS is verified by diffuse reflectance infrared Fourier transform (DRIFT) analysis, and the modified PMHS is shown to be stable when exposed to extreme pH conditions between 2 and 9. Spectrophotometer measurements show that PMHS exhibits over 40% transmittance for ultraviolet (UV) wavelength as low as 220 nm, indicating viability for sensor applications based on UV absorption. The UV transmittance is furthermore observed to be insensitive to thickness for specimens tested between 1.6 mm and 6.4 mm thick. Full curing of PMHS liquid resin occurs between 48 and 72 h at 110 °C with no secondary additives. Casting of microscale features is achieved by using soft lithography methods similar to established techniques for fabrication based on polydimethylsiloxane (PDMS). Microchannels approximately 100 µm wide and 50 µm deep are also demonstrated by carbon dioxide laser ablation, with uniform channels produced using an energy dose of 0.2 mJ mm-1 with respect to line length. Other basic functional requirements for microfluidic chips are discussed, including the ability to bond PMHS substrates by plasma treatment.

  18. A microfluidic chip with hydrodynamic traps for in vitro microscopic investigations of single cells

    NASA Astrophysics Data System (ADS)

    Kukhtevich, I. V.; Belousov, K. I.; Bukatin, A. S.; Dubina, M. V.; Evstrapov, A. A.

    2015-03-01

    The results on making a microfluidic chip for in vitro microscopic investigations of single cells are presented. Numerical simulation of the motion trajectories of microparticles makes it possible to determine the geometry of hydrodynamic traps, their number, and the trap arrangement in a reaction chamber. According to the developed design, microfluidic chips were fabricated from a SU-8 photoresist by photolithography. The microfluidic chips have been tested to prove their operating capacity for isolating and holding K562 human myeloid leukemia cells from a sample flow and their subsequent investigation by confocal laser scanning microscopy.

  19. An integrated, multiparametric flow cytometry chip using "microfluidic drifting" based three-dimensional hydrodynamic focusing.

    PubMed

    Mao, Xiaole; Nawaz, Ahmad Ahsan; Lin, Sz-Chin Steven; Lapsley, Michael Ian; Zhao, Yanhui; McCoy, J Philip; El-Deiry, Wafik S; Huang, Tony Jun

    2012-06-01

    In this work, we demonstrate an integrated, single-layer, miniature flow cytometry device that is capable of multi-parametric particle analysis. The device integrates both particle focusing and detection components on-chip, including a "microfluidic drifting" based three-dimensional (3D) hydrodynamic focusing component and a series of optical fibers integrated into the microfluidic architecture to facilitate on-chip detection. With this design, multiple optical signals (i.e., forward scatter, side scatter, and fluorescence) from individual particles can be simultaneously detected. Experimental results indicate that the performance of our flow cytometry chip is comparable to its bulky, expensive desktop counterpart. The integration of on-chip 3D particle focusing with on-chip multi-parametric optical detection in a single-layer, mass-producible microfluidic device presents a major step towards low-cost flow cytometry chips for point-of-care clinical diagnostics.

  20. 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

  1. PPC750 Performance Monitor

    NASA Technical Reports Server (NTRS)

    Meyer, Donald; Uchenik, Igor

    2007-01-01

    The PPC750 Performance Monitor (Perfmon) is a computer program that helps the user to assess the performance characteristics of application programs running under the Wind River VxWorks real-time operating system on a PPC750 computer. Perfmon generates a user-friendly interface and collects performance data by use of performance registers provided by the PPC750 architecture. It processes and presents run-time statistics on a per-task basis over a repeating time interval (typically, several seconds or minutes) specified by the user. When the Perfmon software module is loaded with the user s software modules, it is available for use through Perfmon commands, without any modification of the user s code and at negligible performance penalty. Per-task run-time performance data made available by Perfmon include percentage time, number of instructions executed per unit time, dispatch ratio, stack high water mark, and level-1 instruction and data cache miss rates. The performance data are written to a file specified by the user or to the serial port of the computer

  2. Integrated circuit/microfluidic chip to programmably trap and move cells and droplets with dielectrophoresis.

    PubMed

    Hunt, Thomas P; Issadore, David; Westervelt, R M

    2008-01-01

    We present an integrated circuit/microfluidic chip that traps and moves individual living biological cells and chemical droplets along programmable paths using dielectrophoresis (DEP). Our chip combines the biocompatibility of microfluidics with the programmability and complexity of integrated circuits (ICs). The chip is capable of simultaneously and independently controlling the location of thousands of dielectric objects, such as cells and chemical droplets. The chip consists of an array of 128 x 256 pixels, 11 x 11 microm(2) in size, controlled by built-in SRAM memory; each pixel can be energized by a radio frequency (RF) voltage of up to 5 V(pp). The IC was built in a commercial foundry and the microfluidic chamber was fabricated on its top surface at Harvard. Using this hybrid chip, we have moved yeast and mammalian cells through a microfluidic chamber at speeds up to 30 microm sec(-1). Thousands of cells can be individually trapped and simultaneously positioned in controlled patterns. The chip can trap and move pL droplets of water in oil, split one droplet into two, and mix two droplets into one. Our IC/microfluidic chip provides a versatile platform to trap and move large numbers of cells and fluid droplets individually for lab-on-a-chip applications.

  3. Lab-chip HPLC with integrated droplet-based microfluidics for separation and high frequency compartmentalisation.

    PubMed

    Kim, Jin-Young; Cho, Soong-Won; Kang, Dong-Ku; Edel, Joshua B; Chang, Soo-Ik; deMello, Andrew J; O'Hare, Danny

    2012-09-21

    We demonstrate the integration of a droplet-based microfluidic device with high performance liquid chromatography (HPLC) in a monolithic format. Sequential operations of separation, compartmentalisation and concentration counter were conducted on a monolithic chip. This describes the use of droplet-based microfluidics for the preservation of chromatographic separations, and its potential application as a high frequency fraction collector.

  4. A world-to-chip socket for microfluidic prototype development

    NASA Astrophysics Data System (ADS)

    Yang, Zhen; Maeda, Ryutaro

    2002-11-01

    This paper reports a prototype for a standard connector between a microfluidic chip and the macro world. This prototype is the first to demonstrate a fully functioning socket for a microchip to access the outside world by means of fluids, data and energy supply, as well as providing process visibility. It has 20 channels for the input and output of liquids or gases, as well as compressed air or vacuum lines for pneumatic power lines. It also contains 42 pins for electrical signals and power. All these connections were designed in a planar configuration with linear orthogonal arrays. The vertical space was opened for optical measurement and evaluation. The die (29.1 mm x 27.5 mm x 0.9 mm) can be easily mounted and dismounted from the socket. No adhesives or solders are used at any contact points. The pressure limit for the connection of working fluids was 0.2 MPa and the current limit for the electrical connections was 1 A. This socket supports both serial and parallel processing applications. It exhibits great potential for developing microfluidic system efficiently.

  5. 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.

  6. A Rapidly Fabricated Microfluidic Chip for Cell Culture.

    PubMed

    Li, Rui; Lv, Xuefei; Hasan, Murtaza; Xu, Jiandong; Xu, Yuanqing; Zhang, Xingjian; Qin, Kuiwei; Wang, Jianshe; Zhou, Di; Deng, Yulin

    2016-04-01

    Microfluidic chips (μFC) are emerging as powerful tools in chemistry, biochemistry, nanotechnology and biotechnology. The microscale size, possibility of integration and high-throughput present huge technical potential to facilitate the research of cell behavior by creating in vivo-like microenvironments. Here, we have developed a new method for rapid fabrication of μFC with Norland Optical Adhesive 81 (NOA81) for multiple cell culture with high efficiency. The proposed method is more suitable for the early structure exploration stage of μFC than existing procedures since no templates are needed and fast fabrication methods are presented. Simple PDMS-NOA81-linked microvalves were embedded in the μFC to control or block the fluid flow effectively, which significantly broadened the applications of μFC. Various types of cells were integrated into the chip and normal viabilities were maintained up to 1 week. Besides, concentration gradient was generated to investigate the cells in the μFC responded to drug stimulation. The cells appeared different in terms of shape and proliferation that strongly demonstrated the potential application of our μFC in online drug delivery. The high biocompatibility of NOA81 and its facile fabrication (μFC) promise its use in various cell analyses, such as cell-cell interactions or tissue engineering.

  7. 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.

  8. PDMS based microfluidic chips and their application in material synthesis

    NASA Astrophysics Data System (ADS)

    Gong, Xiuqing

    reactions. Here, we report the microfluidic fabrication of magnetically responsive microsphere, macroporous polymer microspheres and hollow titania microspheres. To prepare magnetically responsive microsphere, we introduced magnetic particles into liquid shell and drug into liquid core. After cross-linking reaction of the shell, we studied the magnetic contraction and extention behavior which induced the drug release efficiency. To prepare porous polymer, the H 2O2 solution was encapsulated in polymer precursor, after which we investigated its decomposition under UV irradiation, which simultaneously induces the polymerization of the encapsulating shell. Because the H 2O2 decomposition leads to the release of oxygen, porous microspheres were obtained from a combined H2O2-decomposition/polymer precursor polymerization reaction. To prepare hollow titanium gel microspheres, water droplets were first formed by the flow focusing geometry in microfluidic chip and used as a soft template. Then hydrolysis and gelation of titanium alkoxide on the droplet's surface were induced in following serpentine channels, controlled by interface water diffusion. The water diffusion process can be controlled by the amount of the "dewetting" reagent butanol, by which the surface morphology of the titania microspheres can be tuned.

  9. Microfluidic chips for the study of cell migration under the effect of chemicals

    NASA Astrophysics Data System (ADS)

    Kukhtevich, I. V.; Belousov, K. I.; Bukatin, A. S.; Chubinskiy-Nadezhdin, V. I.; Vasileva, V. Yu.; Negulyaev, Yu. A.; Evstrapov, A. A.

    2016-05-01

    Numerical simulation of the formation of a chemoattractant gradient in reaction chambers of a chip having different geometries enabled the determination of a structure suitable for the study of cell migration, in accordance with which hybrid polymer-glass microfluidic devices were manufactured. Verification of the procedures of alignment of cells in the reaction chamber of the chip by centrifugal force and subsequent culturing of the cells showed that microfluidic chips can be used to study cell migration under the effect of the chemoattractant gradient in vitro.

  10. Cooperative Suction by Vertical Capillary Array Pump for Controlling Flow Profiles of Microfluidic Sensor Chips

    PubMed Central

    Horiuchi, Tsutomu; Hayashi, Katsuyoshi; Seyama, Michiko; Inoue, Suzuyo; Tamechika, Emi

    2012-01-01

    A passive pump consisting of integrated vertical capillaries has been developed for a microfluidic chip as an useful component with an excellent flow volume and flow rate. A fluidic chip built into a passive pump was used by connecting the bottoms of all the capillaries to a top surface consisting of a thin layer channel in the microfluidic chip where the thin layer channel depth was smaller than the capillary radius. As a result the vertical capillaries drew fluid cooperatively rather than independently, thus exerting the maximum suction efficiency at every instance. This meant that a flow rate was realized that exhibited little variation and without any external power or operation. A microfluidic chip built into this passive pump had the ability to achieve a quasi-steady rather than a rapidly decreasing flow rate, which is a universal flow characteristic in an ordinary capillary. PMID:23202035

  11. Cooperative suction by vertical capillary array pump for controlling flow profiles of microfluidic sensor chips.

    PubMed

    Horiuchi, Tsutomu; Hayashi, Katsuyoshi; Seyama, Michiko; Inoue, Suzuyo; Tamechika, Emi

    2012-10-18

    A passive pump consisting of integrated vertical capillaries has been developed for a microfluidic chip as an useful component with an excellent flow volume and flow rate. A fluidic chip built into a passive pump was used by connecting the bottoms of all the capillaries to a top surface consisting of a thin layer channel in the microfluidic chip where the thin layer channel depth was smaller than the capillary radius. As a result the vertical capillaries drew fluid cooperatively rather than independently, thus exerting the maximum suction efficiency at every instance. This meant that a flow rate was realized that exhibited little variation and without any external power or operation. A microfluidic chip built into this passive pump had the ability to achieve a quasi-steady rather than a rapidly decreasing flow rate, which is a universal flow characteristic in an ordinary capillary.

  12. Re-use of commercial microfluidics chips for DNA, RNA, and protein electrophoresis.

    PubMed

    Nguyen, Thi; Kwak, Sukyoung; Karpowicz, Steven J

    2014-11-01

    Microfluidics chip technology is a powerful and convenient alternative to agarose gels and PAGE, but costs can be high due to certain chips being non-reusable. Here we describe a method to regenerate, re-use, and store Agilent DNA, RNA, and protein electrophoresis chips designed for use in the Bioanalyzer 2100. By washing the sample wells and displacing the old gel matrix with new gel-dye mix, we have run samples on the same chip up to ten times with negligible loss of signal quality. Chips whose wells were loaded with buffer or water were stored successfully for one week before re-use.

  13. A chemically inert multichannel chip-to-world interface to connect microfluidic chips

    NASA Astrophysics Data System (ADS)

    Neumann, Christiane; Wilhelm, Elisabeth; Duttenhofer, Thomas; Pires, Leonardo; Rapp, Bastian E.

    2014-03-01

    Within the last decades more and more microfluidic systems for applications in chemistry, biology or medicine were developed. Most of them need a connection between the chip and its macroscopic environment e.g., pumps. Numerous concepts for such interconnections are known from literature but most of them allow only a small number of connections and are neither chemically inert nor contamination-free. We developed a chemically inert, reusable, multichannel Chipto- World-Interface (CWI) based on a force fit connection. This principle is comparable to hollow screws as used in highperformance liquid chromatography. The CWI can be used to connect chips, made of different materials, e.g., glass, polydimethylsiloxane (PDMS), or epoxy polymers, with up to 100 thermoplastic tubes. The dimensions of the CWI and the number of connections can be individually adapted depending on the chip dimensions but the pitch between the tubes is fixed. Due to the design of the CWI the fluid is only in contact with the chip and the tubing material, thus leading to a contamination free and zero dead volume interconnection. Using tubes of polytetrafluorethylene (PTFE, Teflon®) even enables probing with organic solvents like dimethylformamide, dichloromethane or tetrahydrofuran over several hours without leakage or corrosion of the CWI. During experiments the CWI with 100 connections resisted pressure up to 630 kPa (6.3 bar) and sustained flow rates higher than 4 ml/min.

  14. Microfluidic cytometers with integrated on-chip optical components for blood cell analysis

    NASA Astrophysics Data System (ADS)

    Zhao, Yingying; Li, Qin; Hu, Xiao-Ming

    2016-10-01

    In the last two decades, microfluidic technologies have shown the great potential in developing portable and point-of care testing blood cell analysis devices. It is challenging to integrate all free-space detecting components in a single microfluidic platform. In this paper, a microfluidic cytometer with integrated on-chip optical components was demonstrated. To facilitate on-chip detection, the device integrated optical fibers and on-chip microlens with microfluidic channels on one polydimethylsiloxane layer by standard soft photolithography. This compact design increased the sensitivity of the device and also eliminated time-consuming free-space optical alignments. Polystyrene particles, together with red blood cells and platelets, were measured in the microfluidic cytometer by small angle forward scatter. Experimental results indicated that the performance of the microfluidic device was comparable to a conventional cytometer. And it was also demonstrated its ability to detect on-chip optical signals in a highly compact, simple, truly portable and low cost format which was perfect suitable for point-of-care testing clinical hematology diagnostics.

  15. Fabrication of 25 μm-filter microfluidic chip on silicon substrate

    NASA Astrophysics Data System (ADS)

    Ngan Le, Nguyen; Khanh Huynh, Kim; Cam Hue Phan, Thi; Dung Dang, Thi My; Chien Dang, Mau

    2017-03-01

    This paper presents the entire fabrication process including photolithography, sputtering, deep reactive ion etching (Bosch DRIE process) on silicon substrate and bonding process between the lid and silicon substrate to create a designed filtration microfluidic chip with dimension of 28 mm × 7 mm, one inlet port and one outlet port. A pattered silver thin film was deposited on a silicon sample by the lift-off method. Subsequently the newly fabricated sample was anisotropically etched by Bosch DRIE process. Some parameters of Bosch DRIE process such as bias power, duration of etching step and passivation step, oxygen presence were studied to explore the dependence of silicon channel depth and etched shape profile on these parameters. An optimized process was utilized to fabricate a featured silicon channel with vertical, smooth sidewalls and an overall good uniformity. The silicon channel has four arrays of microposts with various distances between microposts from 25 μm to 100 μm. The depth of the silicon channel was about 150 μm. After that, silicon substrate was bonded with mica lid by adhesive bonding method to form the completed filtration microfluidic chip. The samples were characterized by scanning electron microscopy (SEM), mechanical profilometer (DEKTAK 6 M), optical microscopy (Olympus MX51). In this paper a test was performed to demonstrate how the microfluidic chip works by pumping solution with many various sizes of particles through the inlet port of the microfluidic chip and obtaining a solution with desired particles sizes (smaller than 25 μm) through another port. Moreover, the chip could be pumped de-ionized water through outlet port for backwash in order to make this microfluidic chip reusable. Finally, a few applications of microfluidic chips are presented to illustrate the advantages of this technology and the potential for future development. Invited talk at 8th International Workshop on Advanced Materials Science and Nanotechnology

  16. Rapid fabrication of a four-layer PMMA-based microfluidic chip using CO2-laser micromachining and thermal bonding

    NASA Astrophysics Data System (ADS)

    Chen, Xueye; Shen, Jienan; Zhou, Mengde

    2016-10-01

    A smart design method to transform the original two-layer microfluidic chip into a four-layer 3D microfluidic chip is proposed. A novel fabrication method is established to rapidly and effectively produce a four-layer microfluidic chip device made entirely from polymethylmethacrylate (PMMA). Firstly, the CO2-laser cuts the PMMA sheets by melting and blowing away vaporized material from the parent material to obtain high-quality channels of the microfluidic chip. An orthogonal experimental method is used to study its processing stability. In addition, a simple, rapid thermal bonding technique is successfully applied in fabricating the four-layer microfluidic chip, which has a bond strength of 1.3 MPa. A wooden pole is used to improve the accuracy of the alignment. Finally, a mixing experiment with blue ink and water is carried out, which proves that this smart design method and rapid manufacturing technology are successful.

  17. On-chip microfluidic biosensor using superparamagnetic microparticles

    PubMed Central

    Kokkinis, G.; Keplinger, F.; Giouroudi, I.

    2013-01-01

    In this paper, an integrated solution towards an on-chip microfluidic biosensor using the magnetically induced motion of functionalized superparamagnetic microparticles (SMPs) is presented. The concept of the proposed method is that the induced velocity on SMPs in suspension, while imposed to a magnetic field gradient, is inversely proportional to their volume. Specifically, a velocity variation of suspended functionalized SMPs inside a detection microchannel with respect to a reference velocity, specified in a parallel reference microchannel, indicates an increase in their non-magnetic volume. This volumetric increase of the SMPs is caused by the binding of organic compounds (e.g., biomolecules) to their functionalized surface. The new compounds with the increased non-magnetic volume are called loaded SMPs (LSMPs). The magnetic force required for the manipulation of the SMPs and LSMPs is produced by current currying conducting microstructures, driven by a programmable microcontroller. Experiments were carried out as a proof of concept. A promising decrease in the velocity of the LSMPs in comparison to that of the SMPs was measured. Thus, it is the velocity variation which determines the presence of the organic compounds in the sample fluid. PMID:24396528

  18. Microfluidic on-chip fluorescence-activated interface control system.

    PubMed

    Haiwang, Li; Nguyen, N T; Wong, T N; Ng, S L

    2010-11-22

    A microfluidic dynamic fluorescence-activated interface control system was developed for lab-on-a-chip applications. The system consists of a straight rectangular microchannel, a fluorescence excitation source, a detection sensor, a signal conversion circuit, and a high-voltage feedback system. Aqueous NaCl as conducting fluid and aqueous glycerol as nonconducting fluid were introduced to flow side by side into the straight rectangular microchannel. Fluorescent dye was added to the aqueous NaCl to work as a signal representing the interface position. Automatic control of the liquid interface was achieved by controlling the electroosmotic effect that exists only in the conducting fluid using a high-voltage feedback system. A LABVIEW program was developed to control the output of high-voltage power supply according the actual interface position, and then the interface position is modified as the output of high-voltage power supply. At last, the interface can be moved to the desired position automatically using this feedback system. The results show that the system presented in this paper can control an arbitrary interface location in real time. The effects of viscosity ratio, flow rates, and polarity of electric field were discussed. This technique can be extended to switch the sample flow and droplets automatically.

  19. Direct optical patterning of poly(dimethylsiloxane) microstructures for microfluidic chips

    NASA Astrophysics Data System (ADS)

    Gao, Shaorui; Tung, Wing-Tai; Wong, Dexter S.; Bian, Liming; Zhang, A. Ping

    2016-10-01

    In this paper, we present an optical maskless exposure approach for direct patterning of large-area high resolution microfluidic chips using photosensitive poly(dimethylsiloxane) (PDMS) materials. Both positive- and negative-tone photosensitive PDMS (photoPDMS) were successfully patterned into various microfluidic devices with complex geometries by using an optical maskless lithography process. The positive-tone PDMS is used for patterning of largearea chips, while the negative-tone PDMS is demonstrated to fabricate high-resolution microstructures and on-chip devices. With the seamless pattern-stitching technique, a large-area microfluidic chip with size of 5.5 × 2.8 cm2 with complex three-dimensional (3D) staggered herringbone mixers (SHMs) for micro-flow gradient generation has been directly fabricated within 125 minutes by using the positive-tone PDMS. A small microfluidic chip with feature size as small as 5 μm is demonstrated by using the negative-tone PDMS. The experimental results reveal that the optical maskless lithography technology enables to rapidly pattern high-resolution microstructures and is very promising for development of lab-on-a-chip devices.

  20. Thin film metal sensors in fusion bonded glass chips for high-pressure microfluidics

    NASA Astrophysics Data System (ADS)

    Andersson, Martin; Ek, Johan; Hedman, Ludvig; Johansson, Fredrik; Sehlstedt, Viktor; Stocklassa, Jesper; Snögren, Pär; Pettersson, Victor; Larsson, Jonas; Vizuete, Olivier; Hjort, Klas; Klintberg, Lena

    2017-01-01

    High-pressure microfluidics offers fast analyses of thermodynamic parameters for compressed process solvents. However, microfluidic platforms handling highly compressible supercritical CO2 are difficult to control, and on-chip sensing would offer added control of the devices. Therefore, there is a need to integrate sensors into highly pressure tolerant glass chips. In this paper, thin film Pt sensors were embedded in shallow etched trenches in a glass wafer that was bonded with another glass wafer having microfluidic channels. The devices having sensors integrated into the flow channels sustained pressures up to 220 bar, typical for the operation of supercritical CO2. No leakage from the devices could be found. Integrated temperature sensors were capable of measuring local decompression cooling effects and integrated calorimetric sensors measured flow velocities over the range 0.5-13.8 mm s-1. By this, a better control of high-pressure microfluidic platforms has been achieved.

  1. IFSA: a microfluidic chip-platform for frit-based immunoassay protocols

    NASA Astrophysics Data System (ADS)

    Hlawatsch, Nadine; Bangert, Michael; Miethe, Peter; Becker, Holger; Gärtner, Claudia

    2013-03-01

    Point-of-care diagnostics (POC) is one of the key application fields for lab-on-a-chip devices. While in recent years much of the work has concentrated on integrating complex molecular diagnostic assays onto a microfluidic device, there is a need to also put comparatively simple immunoassay-type protocols on a microfluidic platform. In this paper, we present the development of a microfluidic cartridge using an immunofiltration approach. In this method, the sandwich immunoassay takes place in a porous frit on which the antibodies have immobilized. The device is designed to be able to handle three samples in parallel and up to four analytical targets per sample. In order to meet the critical cost targets for the diagnostic market, the microfluidic chip has been designed and manufactured using high-volume manufacturing technologies in mind. Validation experiments show comparable sensitivities in comparison with conventional immunofiltration kits.

  2. Development of a Plastic-Based Microfluidic Immunosensor Chip for Detection of H1N1 Influenza

    PubMed Central

    Lee, Kyoung G.; Lee, Tae Jae; Jeong, Soon Woo; Choi, Ho Woon; Heo, Nam Su; Park, Jung Youn; Park, Tae Jung; Lee, Seok Jae

    2012-01-01

    Lab-on-a-chip can provide convenient and accurate diagnosis tools. In this paper, a plastic-based microfluidic immunosensor chip for the diagnosis of swine flu (H1N1) was developed by immobilizing hemagglutinin antigen on a gold surface using a genetically engineered polypeptide. A fluorescent dye-labeled antibody (Ab) was used for quantifying the concentration of Ab in the immunosensor chip using a fluorescent technique. For increasing the detection efficiency and reducing the errors, three chambers and three microchannels were designed in one microfluidic chip. This protocol could be applied to the diagnosis of other infectious diseases in a microfluidic device. PMID:23112630

  3. Integration of Curved D-Type Optical Fiber Sensor with Microfluidic Chip

    PubMed Central

    Sun, Yung-Shin; Li, Chang-Jyun; Hsu, Jin-Cherng

    2016-01-01

    A curved D-type optical fiber sensor (OFS) combined with a microfluidic chip is proposed. This OFS, based on surface plasmon resonance (SPR) of the Kretchmann’s configuration, is applied as a biosensor to measure the concentrations of different bio-liquids such as ethanol, methanol, and glucose solutions. The SPR phenomenon is attained by using the optical fiber to guide the light source to reach the side-polished, gold-coated region. Integrating this OFS with a polymethylmethacrylate (PMMA)-based microfluidic chip, the SPR spectra for liquids with different refractive indices are recorded. Experimentally, the sensitivity of the current biosensor was calculated to be in the order of 10−5 RIU. This microfluidic chip-integrated OFS could be valuable for monitoring subtle changes in biological samples such as blood sugar, allergen, and biomolecular interactions. PMID:28042821

  4. Rapid fabrication of microfluidic chips based on the simplest LED lithography

    NASA Astrophysics Data System (ADS)

    Li, Yue; Wu, Ping; Luo, Zhaofeng; Ren, Yuxuan; Liao, Meixiang; Feng, Lili; Li, Yuting; He, Liqun

    2015-05-01

    Microfluidic chips are generally fabricated by a soft lithography method employing commercial lithography equipment. These heavy machines require a critical room environment and high lamp power, and the cost remains too high for most normal laboratories. Here we present a novel microfluidics fabrication method utilizing a portable ultraviolet (UV) LED as an alternative UV source for photolithography. With this approach, we can repeat several common microchannels as do these conventional commercial exposure machines, and both the verticality of the channel sidewall and lithography resolution are proved to be acceptable. Further microfluidics applications such as mixing, blood typing and microdroplet generation are implemented to validate the practicability of the chips. This simple but innovative method decreases the cost and requirement of chip fabrication dramatically and may be more popular with ordinary laboratories.

  5. 3D-printed microfluidic chips with patterned, cell-laden hydrogel constructs.

    PubMed

    Knowlton, Stephanie; Yu, Chu Hsiang; Ersoy, Fulya; Emadi, Sharareh; Khademhosseini, Ali; Tasoglu, Savas

    2016-06-20

    Three-dimensional (3D) printing offers potential to fabricate high-throughput and low-cost fabrication of microfluidic devices as a promising alternative to traditional techniques which enables efficient design iterations in the development stage. In this study, we demonstrate a single-step fabrication of a 3D transparent microfluidic chip using two alternative techniques: a stereolithography-based desktop 3D printer and a two-step fabrication using an industrial 3D printer based on polyjet technology. This method, compared to conventional fabrication using relatively expensive materials and labor-intensive processes, presents a low-cost, rapid prototyping technique to print functional 3D microfluidic chips. We enhance the capabilities of 3D-printed microfluidic devices by coupling 3D cell encapsulation and spatial patterning within photocrosslinkable gelatin methacryloyl (GelMA). The platform presented here serves as a 3D culture environment for long-term cell culture and growth. Furthermore, we have demonstrated the ability to print complex 3D microfluidic channels to create predictable and controllable fluid flow regimes. Here, we demonstrate the novel use of 3D-printed microfluidic chips as controllable 3D cell culture environments, advancing the applicability of 3D printing to engineering physiological systems for future applications in bioengineering.

  6. Active pneumatic control of centrifugal microfluidic flows for lab-on-a-chip applications.

    PubMed

    Clime, Liviu; Brassard, Daniel; Geissler, Matthias; Veres, Teodor

    2015-06-07

    This paper reports a novel method of controlling liquid motion on a centrifugal microfluidic platform based on the integration of a regulated pressure pump and a programmable electromechanical valving system. We demonstrate accurate control over the displacement of liquids within the system by pressurizing simultaneously multiple ports of the microfluidic device while the platform is rotating at high speed. Compared to classical centrifugal microfluidic platforms where liquids are solely driven by centrifugal and capillary forces, the method presented herein adds a new degree of freedom for fluidic manipulation, which represents a paradigm change in centrifugal microfluidics. We first demonstrate how various core microfluidic functions such as valving, switching, and reverse pumping (i.e., against the centrifugal field) can be easily achieved by programming the pressures applied at dedicated access ports of the microfluidic device. We then show, for the first time, that the combination of centrifugal force and active pneumatic pumping offers the possibility of mixing fluids rapidly (~0.1 s) and efficiently based on the creation of air bubbles at the bottom of a microfluidic reservoir. Finally, the suitability of the developed platform for performing complex bioanalytical assays in an automated fashion is demonstrated in a DNA harvesting experiment where recovery rates of about 70% were systematically achieved. The proposed concept offers the interesting prospect to decouple basic microfluidic functions from specific material properties, channel dimensions and fabrication tolerances, surface treatments, or on-chip active components, thus promoting integration of complex assays on simple and low-cost microfluidic cartridges.

  7. Microfluidic-integrated laser-controlled microactuators with on-chip microscopy imaging functionality

    PubMed Central

    Jung, Jae Hee; Han, Chao; Lee, Seung Ah; Kim, Jinho; Yang, Changhuei

    2014-01-01

    The fabrication of a novel microfluidic system, integrated with a set of laser-controlled microactuators on an ePetri on-chip microscopy platform, is presented in this paper. In the fully integrated microfluidic system, a set of novel thermally actuated paraffin-based microactuators, precisely controlled by programmed laser optics, was developed to regulate flow and to provide pumping of liquid solutions without external connections. The microfluidic chip was fabricated on a complementary metal–oxide–semiconductor (CMOS)-imaging sensor chip on an ePetri platform; this configuration provided real-time, wide field-of-view, high-resolution imaging using a sub-pixel sweeping microscopy technique. The system of microactuators, which consisted of microvalves and a micropump, operated well in the microfluidic channel with a focused near-infrared laser beam providing the actuation control. As a demonstration, we used our prototype to assess cell–drug interactions, and monitored cell growth directly within an incubator in real time. The powerful combination of the laser-actuated microfluidics and chip-scale microscopy techniques represents a significant step forward in terms of a simple, robust, high-throughput, and highly compact analysis system for biomedical and bioscience applications. PMID:25099225

  8. Design, fabrication and test of a microfluidic nebulizer chip for desorption electrospray ionization mass spectrometry

    PubMed Central

    Sen, A K; Darabi, J; Knapp, D R

    2009-01-01

    This paper presents design, microfabrication, and test of a microfluidic nebulizer chip for desorption electrospray ionization mass spectrometry (DESI-MS) in proteomic analysis. The microfluidic chip is fabricated using cyclic olefin copolymer (COC) substrates. The fluidic channels are thermally embossed onto a base substrate using a nickel master and then a top substrate is thermally bonded to seal the channels. Carbon ink embossed into the top COC substrate is used to established electrical connection between the external power supply and the liquid in the channel. The microfluidic chip to external capillary connection is fabricated using Nanoport™ interconnection system. Preliminary leakage test was performed to demonstrate the interconnection system is leak-free and pressure test was performed to evaluate the burst pressure. Finally, the nebulizer chip was used to perform DESI-MS for analyzing peptides (BSA and bradykinin) and reserpine on the nanoporous alumina surface. DESI-MS performance of the microfluidic nebulizer chip is compared with that obtained using a conventional DESI nebulizer. PMID:20161284

  9. Manually operatable on-chip bistable pneumatic microstructures for microfluidic manipulations.

    PubMed

    Chen, Arnold; Pan, Tingrui

    2014-09-07

    Bistable microvalves are of particular interest because of their distinct nature of requiring energy consumption only during the transition between the open and closed states. This characteristic can be highly advantageous in reducing the number of external inputs and the complexity of control circuitries since microfluidic devices as contemporary lab-on-a-chip platforms are transferring from research settings to low-resource environments with high integrability and a small form factor. In this paper, we first present manually operatable, on-chip bistable pneumatic microstructures (BPMs) for microfluidic manipulation. The structural design and operation of the BPM devices can be readily integrated into any pneumatically powered microfluidic network consisting of pneumatic and fluidic channels. It is mainly composed of a vacuum activation chamber (VAC) and a pressure release chamber (PRC), of which users have direct control through finger pressing to switch either to the bistable vacuum state (VS) or the atmospheric state (AS). We have integrated multiple BPM devices into a 4-to-1 microfluidic multiplexor to demonstrate on-chip digital flow switching from different sources. Furthermore, we have shown its clinical relevance in a point-of-care diagnostic chip that processes blood samples to identify the distinct blood types (A/B/O) on-chip.

  10. Fabrication of dielectrophoretic microfluidic chips using a facile screen-printing technique for microparticle trapping

    NASA Astrophysics Data System (ADS)

    Wee, Wei Hong; Li, Zedong; Hu, Jie; Adib Kadri, Nahrizul; Xu, Feng; Li, Fei; Pingguan-Murphy, Belinda

    2015-10-01

    Trapping of microparticles finds wide applications in numerous fields. Microfluidic chips based on a dielectrophoresis (DEP) technique hold several advantages for trapping microparticles, such as fast result processing, a small amount of sample required, high spatial resolution, and high accuracy of target selection. There is an unmet need to develop DEP microfluidic chips on different substrates for different applications in a low cost, facile, and rapid way. This study develops a new facile method based on a screen-printing technique for fabrication of electrodes of DEP chips on three types of substrates (i.e. polymethyl-methacrylate (PMMA), poly(ethylene terephthalate) and A4 paper). The fabricated PMMA-based DEP microfluidic chip was selected as an example and successfully used to trap and align polystyrene microparticles in a suspension and cardiac fibroblasts in a cell culture solution. The developed electrode fabrication method is compatible with different kinds of DEP substrates, which could expand the future application field of DEP microfluidic chips, including new forms of point-of care diagnostics and trapping circulating tumor cells.

  11. Manually Operatable On-Chip Bistable Pneumatic Microstructures for Microfluidic Manipulations

    PubMed Central

    Chen, A.; Pan, T.

    2014-01-01

    Bistable microvalves are of particular interest because of their distinct nature requiring energy consumption only during the transition between the open and closed states. This characteristic can be highly advantageous in reducing the number of external inputs and the complexity of control circuitries for microfluidic devices as contemporary lab-on-a-chip platforms are transferring from research settings to low-resource environments with high integratability and small form factor. In this paper, we first present manually operatable, on-chip bistable pneumatic microstructures (BPM) for microfluidic manipulation. The structural design and operation of the BPM devices can be readily integrated into any pneumatically powered microfluidic network consisting of pneumatic and fluidic channels. It is mainly comprised of a vacuum activation chamber (VAC) and a pressure release chamber (PRC), which users have direct control through finger pressing to switch between bistable vacuum state (VS) or atmospheric state (AS). We have integrated multiple BPM devices into a 4-to-1 microfluidic multiplexor to demonstrate on-chip digital flow switching from different sources. Furthermore, we have shown its clinical relevance in a point-of-care diagnostic chip that process blood samples to identify the distinct blood types (A/B/O) on chip. PMID:25007840

  12. Chip-olate’ and dry-film resists for efficient fabrication, singulation and sealing of microfluidic chips

    NASA Astrophysics Data System (ADS)

    Temiz, Yuksel; Delamarche, Emmanuel

    2014-09-01

    This paper describes a technique for high-throughput fabrication and efficient singulation of chips having closed microfluidic structures and takes advantage of dry-film resists (DFRs) for efficient sealing of capillary systems. The technique is illustrated using 4-inch Si/SiO2 wafers. Wafers carrying open microfluidic structures are partially diced to about half of their thickness. Treatments such as surface cleaning are done at wafer-level, then the structures are sealed using low-temperature (45 °C) lamination of a DFR that is pre-patterned using a craft cutter, and ready-to-use chips are finally separated manually like a chocolate bar by applying a small force (≤ 4 N). We further show that some DFRs have low auto-fluorescence at wavelengths typically used for common fluorescent dyes and that mechanical properties of some DFRs allow for the lamination of 200 μm wide microfluidic structures with negligible sagging (~1 μm). The hydrophilicity (advancing contact angle of ~60°) of the DFR supports autonomous capillary-driven flow without the need for additional surface treatment of the microfluidic chips. Flow rates from 1 to 5 µL min-1 are generated using different geometries of channels and capillary pumps. In addition, the ‘chip-olate’ technique is compatible with the patterning of capture antibodies on DFR for use in immunoassays. We believe this technique to be applicable to the fabrication of a wide range of microfluidic and lab-on-a-chip devices and to offer a viable alternative to many labor-intensive processes that are currently based on wafer bonding techniques or on the molding of poly(dimethylsiloxane) (PDMS) layers.

  13. Ex Situ Integration of Multifunctional Porous Polymer Monoliths into Thermoplastic Microfluidic Chips

    PubMed Central

    Kendall, Eric L.; Wienhold, Erik; Rahmanian, Omid D.; DeVoe, Don L.

    2014-01-01

    A unique method for incorporating functional porous polymer monolith elements into thermoplastic microfluidic chips is described. Monolith elements are formed in a microfabricated mold, rather than within the microchannels, and chemically functionalized off chip before insertion into solvent-softened thermoplastic microchannels during chip assembly. Because monoliths may be trimmed prior to final placement, control of their size, shape, and uniformity is greatly improved over in-situ photopolymerization methods. A characteristic trapezoidal profile facilitates rapid insertion and enables complete mechanical anchoring of the monolith periphery, eliminating the need for chemical attachment to the microchannel walls. Off-chip processing allows the parallel preparation of monoliths of differing compositions and surface chemistries in large batches. Multifunctional flow-through arrays of multiple monolith elements are demonstrated using this approach through the creation of a fluorescent immunosensor with integrated controls, and a microfluidic bubble separator comprising a combination of integrated hydrophobic and hydrophilic monolith elements. PMID:25018587

  14. 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.

  15. A microfluidic approach for protein structure determination at room temperature via on-chip anomalous diffraction.

    PubMed

    Perry, Sarah L; Guha, Sudipto; Pawate, Ashtamurthy S; Bhaskarla, Amrit; Agarwal, Vinayak; Nair, Satish K; Kenis, Paul J A

    2013-08-21

    We report a microfluidic approach for de novo protein structure determination via crystallization screening and optimization, as well as on-chip X-ray diffraction data collection. The structure of phosphonoacetate hydrolase (PhnA) has been solved to 2.11 Åvia on-chip collection of anomalous data that has an order of magnitude lower mosaicity than what is typical for traditional structure determination methods.

  16. 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.

  17. Maskless fabrication of cell-laden microfluidic chips with localized surface functionalization for the co-culture of cancer cells.

    PubMed

    Hamid, Qudus; Wang, Chengyang; Snyder, Jessica; Williams, Shannon; Liu, Yigong; Sun, Wei

    2015-03-02

    The utilization of the microfabrication technique to fabricate advanced computing chips has exponentially increased in the last few decades. Needless to say, this fabrication technique offers some unique advantages to develop micro-systems. Though many conventional microfabrication techniques today uses very harsh chemicals, the authors believe that the manipulation of system components and fabrication methods may aid in the utilization of the microfabrication techniques used in fabricating computer chips to develop advanced biological microfluidic systems. Presented in this paper is a fabrication approach in which popular fabrication methods and techniques are coupled together to develop an integrated system that aids in the fabrication of cell-laden microfluidic systems. This system aims to reduce the uses of harsh chemicals and decreases the lengthy fabrication time. Additionally, this integrated system will enable the printing of cells as the microfluidic chip is being fabricated. To demonstrate the unique capabilities of the integrated system, an advanced microfluidic chip is being fabricated and investigated. The advanced chip will feature the investigation of cancer cells in a co-cultured microfluidic environment. The investigations presented demonstrate co-cultures in a microfluidic chip, advanced cell printing with localized surface enhancement, cell integration, and full additive fabrication of a microfluidic chip.

  18. Numerical design of microfluidic-microelectric hybrid chip for the separation of biological cells.

    PubMed

    Ye, Ting; Li, Hua; Lam, K Y

    2011-03-15

    A miniature microfluidic-microelectric hybrid chip is numerically designed for separation of biological cells, where the characteristic length of the chip is close to the cell radius. A mathematical model is developed to characterize the motion and deformation of a biological cell in the hydrodynamic and nonuniform electric coupled fields, in which the mechanical and dielectric behaviors of the cell are taken into consideration. Subsequently, the model is validated by comparing with the experimental results published previously. By taking a red blood cell (RBC) as the sample of biological cell, the chip structure is numerically designed from the viewpoints of the electrode width, fluid flow velocity, and electric potential, respectively. Using the designed microfluidic-microelectric hybrid chip, the effects of the shape and initial position of the RBC on the separation ability are then analyzed. After that, the separation of the RBCs with the different permittivities or conductivities using the designed chip is simulated, and the deformation behaviors of the RBCs are discussed as well. At the high frequency, the permittivities of the RBCs play a dominant role in the separation of the RBCs, which causes the RBCs moving toward or away from the electrode array. However, the conductivity of the RBC plays a significant role at the low frequency. With suitable suspending fluid therefore, the separation of cells with different permittivities or conductivities can be achieved using the microfluidic-microelectric hybrid chip designed by the present work.

  19. Fuel cell-powered microfluidic platform for lab-on-a-chip applications.

    PubMed

    Esquivel, Juan Pablo; Castellarnau, Marc; Senn, Tobias; Löchel, Bernd; Samitier, Josep; Sabaté, Neus

    2012-01-07

    The achievement of a higher degree of integration of components--especially micropumps and power sources--is a challenge currently being pursued to obtain portable and totally autonomous microfluidic devices. This paper presents the integration of a micro direct methanol fuel cell (μDMFC) in a microfluidic platform as a smart solution to provide both electrical and pumping power to a Lab-on-a-Chip system. In this system the electric power produced by the fuel cell is available to enable most of the functionalites required by the microfluidic chip, while the generated CO(2) from the electrochemical reaction produces a pressure capable of pumping a liquid volume through a microchannel. The control of the fuel cell operating conditions allows regulation of the flow rate of a liquid sample through a microfluidic network. The relation between sample flow rate and the current generated by the fuel cell is practically linear, achieving values in the range of 4-18 μL min(-1) while having an available power between 1-4 mW. This permits adjusting the desired flow rate for a given application by controlling the fuel cell output conditions and foresees a fully autonomous analytical Lab-on-a-Chip in which the same device would provide the electrical power to a detection module and at the same time use the CO(2) pumping action to flow the required analytes through a particular microfluidic design.

  20. 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.

  1. A hybrid paper and microfluidic chip with electrowetting valves and colorimetric detection.

    PubMed

    He, Fei; Grimes, Jeff; Alcaine, Samuel D; Nugen, Sam R

    2014-06-21

    Sequential fluid delivery with minimized external equipment is vital towards a point-of-care diagnostic device. In this work, we have further developed the On-chip Electrowetting Valves concept for the sequential delivery of the reagents to the reaction site in a miniaturized capillary-driven microfluidic chip. Specifically, a disposable polymeric microfluidic device was developed containing capillary force driven microchannels. The device was fabricated using laser ablation and inkjet printing and required no external pumping equipment. The assay was conducted on the microchip containing microfluidic channels with embedded electrowetting valves and a porous membrane patterned with capture molecules and colloidal gold labels. To conduct the assay, the microchip was connected with a low voltage supply which was capable of sequentially opening the valves, delivering the sample and the rinsing reagent to generate visual results. Using T7 bacteriophage as a model, we have demonstrated the development of the device, operation of the valves and execution of the automated assay.

  2. Chip in a lab: Microfluidics for next generation life science research

    PubMed Central

    Streets, Aaron M.; Huang, Yanyi

    2013-01-01

    Microfluidic circuits are characterized by fluidic channels and chambers with a linear dimension on the order of tens to hundreds of micrometers. Components of this size enable lab-on-a-chip technology that has much promise, for example, in the development of point-of-care diagnostics. Micro-scale fluidic circuits also yield practical, physical, and technological advantages for studying biological systems, enhancing the ability of researchers to make more precise quantitative measurements. Microfluidic technology has thus become a powerful tool in the life science research laboratory over the past decade. Here we focus on chip-in-a-lab applications of microfluidics and survey some examples of how small fluidic components have provided researchers with new tools for life science research. PMID:23460772

  3. Microfluidic chips for point-of-care immunodiagnostics.

    PubMed

    Gervais, Luc; de Rooij, Nico; Delamarche, Emmanuel

    2011-06-24

    We might be at the turning point where research in microfluidics undertaken in academia and industrial research laboratories, and substantially sponsored by public grants, may provide a range of portable and networked diagnostic devices. In this Progress Report, an overview on microfluidic devices that may become the next generation of point-of-care (POC) diagnostics is provided. First, we describe gaps and opportunities in medical diagnostics and how microfluidics can address these gaps using the example of immunodiagnostics. Next, we conceptualize how different technologies are converging into working microfluidic POC diagnostics devices. Technologies are explained from the perspective of sample interaction with components of a device. Specifically, we detail materials, surface treatment, sample processing, microfluidic elements (such as valves, pumps, and mixers), receptors, and analytes in the light of various biosensing concepts. Finally, we discuss the integration of components into accurate and reliable devices.

  4. "Print-n-Shrink" technology for the rapid production of microfluidic chips and protein microarrays.

    PubMed

    Sollier, Kevin; Mandon, Céline A; Heyries, Kevin A; Blum, Loïc J; Marquette, Christophe A

    2009-12-21

    An innovative method for the production of microfluidic chips integrating protein spots is described. The technology, called "Print-n-Shrink", is based on the screen-printing of a microfluidic design (using a dielectric ink) onto Polyshrink polystyrene sheets. The initial print which has a minimum size of 15 microm (height) x 230 microm (width) is thermally treated (30 seconds, 163 degrees C) to shrink and generate features of 85 microm (height) x 100 microm (width). Concomitantly, proteins such as monoclonal antibodies or cellular adhesion proteins are spotted onto the Polyshrink sheets and shrunk together with the microfluidic design, creating a complete biochip integrating both complex microfluidic designs and protein spots for bioanalytical applications.

  5. Rapid detection of pathogens using antibody-coated microbeads with bioluminescence in microfluidic chips.

    PubMed

    Guan, Xiao; Zhang, Hui-jing; Bi, Yin-nan; Zhang, Li; Hao, Dun-ling

    2010-08-01

    Detection of pathogens was demonstrated in a polydimethylsiloxane (PDMS)/glass microfluidic chip with which microbead-based immunoseparation platform and the bioluminescence technology were integrated. Escherichia coli (E. coli) O157:H7 was used as the model bacteria. The microchamber in microfluidic chip was filled with glass beads coated with antibodies which could capture specific organism, and the capture efficiency of the chip for the bacteria was about 91.75% approximately 95.62%. Then the concentration of bacteria was determined by detecting adenosine triphosphate (ATP) employing bioluminescence reaction of firefly luciferin-lucifera-ATP on chip. The method allowed reliable detection of E. coli O157:H7 concentrations from 3.2 x 10(1) cfu/microL to 3.2 x 10(5) cfu/microL within 20 min. This research demonstrated excellent reproducibility, stability, and specificity, and could accurately detect the pathogenic bacteria in food samples. The microfluidic chip and the equipments used in this method are easy to miniaturize, thus the method has great potential to be developed to a portable device for rapid detection of pathogens.

  6. Microfluidic Gut-liver chip for reproducing the first pass metabolism.

    PubMed

    Choe, Aerim; Ha, Sang Keun; Choi, Inwook; Choi, Nakwon; Sung, Jong Hwan

    2017-03-01

    After oral intake of drugs, drugs go through the first pass metabolism in the gut and the liver, which greatly affects the final outcome of the drugs' efficacy and side effects. The first pass metabolism is a complex process involving the gut and the liver tissue, with transport and reaction occurring simultaneously at various locations, which makes it difficult to be reproduced in vitro with conventional cell culture systems. In an effort to tackle this challenge, here we have developed a microfluidic gut-liver chip that can reproduce the dynamics of the first pass metabolism. The microfluidic chip consists of two separate layers for gut epithelial cells (Caco-2) and the liver cells (HepG2), and is designed so that drugs go through a sequential absorption in the gut chamber and metabolic reaction in the liver chamber. We fabricated the chip and showed that the two different cell lines can be successfully co-cultured on chip. When the two cells are cultured on chip, changes in the physiological function of Caco-2 and HepG2 cells were noted. The cytochrome P450 metabolic activity of both cells were significantly enhanced, and the absorptive property of Caco-2 cells on chip also changed in response to the presence of flow. Finally, first pass metabolism of a flavonoid, apigenin, was evaluated as a model compound, and co-culture of gut and liver cells on chip resulted in a metabolic profile that is closer to the reported profile than a monoculture of gut cells. This microfluidic gut-liver chip can potentially be a useful platform to study the complex first pass metabolism of drugs in vitro.

  7. Direct-writing colloidal photonic crystal microfluidic chips by inkjet printing for label-free protein detection.

    PubMed

    Shen, Weizhi; Li, Mingzhu; Ye, Changqing; Jiang, Lei; Song, Yanlin

    2012-09-07

    Integrating photonic crystals (PC) into microfluidic systems has attracted immense interest for its novel functions. However, it is still a great challenge to fabricate PC microfluidic chips rapidly with complex functions. In this work, a direct-writing colloidal PC microchannel was firstly achieved by inkjet printing and was used for the surface-tension-confined microfluidic immune assay. PC channels with different structure colors have been successfully integrated on one chip. The fabricated chip has the advantages of rapid fabrication, quick fluidic transport and can monitor the fluidic fluxion using the naked eye. Utilizing this PC microfluidic chip, a colorimetric label-free immune assay was realized without nonspecific adsorption interference of the target.

  8. Digital Microfluidic Dynamic Culture of Mammalian Embryos on an Electrowetting on Dielectric (EWOD) Chip

    PubMed Central

    Huang, Hong-Yuan; Shen, Hsien-Hua; Tien, Chang-Hung; Li, Chin-Jung; Fan, Shih-Kang; Liu, Cheng-Hsien; Hsu, Wen-Syang; Yao, Da-Jeng

    2015-01-01

    Current human fertilization in vitro (IVF) bypasses the female oviduct and manually inseminates, fertilizes and cultivates embryos in a static microdrop containing appropriate chemical compounds. A microfluidic microchannel system for IVF is considered to provide an improved in-vivo-mimicking environment to enhance the development in a culture system for an embryo before implantation. We demonstrate a novel digitalized microfluidic device powered with electrowetting on a dielectric (EWOD) to culture an embryo in vitro in a single droplet in a microfluidic environment to mimic the environment in vivo for development of the embryo and to culture the embryos with good development and live births. Our results show that the dynamic culture powered with EWOD can manipulate a single droplet containing one mouse embryo and culture to the blastocyst stage. The rate of embryo cleavage to a hatching blastocyst with a dynamic culture is significantly greater than that with a traditional static culture (p<0.05). The EWOD chip enhances the culture of mouse embryos in a dynamic environment. To test the reproductive outcome of the embryos collected from an EWOD chip as a culture system, we transferred embryos to pseudo-pregnant female mice and produced live births. These results demonstrate that an EWOD-based microfluidic device is capable of culturing mammalian embryos in a microfluidic biological manner, presaging future clinical application. PMID:25933003

  9. Capillary-driven microfluidic chips with evaporation-induced flow control and dielectrophoretic microbead trapping

    NASA Astrophysics Data System (ADS)

    Temiz, Yuksel; Skorucak, Jelena; Delamarche, Emmanuel

    2014-07-01

    This work reports our efforts on developing simple-to-use microfluidic devices for point-of-care diagnostic applications with recent extensions that include the trapping of microbeads using dielectrophoresis (DEP) and the modulation of the liquid flow using integrated microheaters. DEP serves the purpose of trapping microbeads coated with receptors and analytes for detection of a fluorescent signal. The microheater is actuated once the chip is filled by capillarity, creating an evaporation-induced flow tuned according to assay conditions. The chips are composed of a glass substrate patterned with 50-nm-thick Pd electrodes and microfluidic structures made using a 20-μm-thick dry-film resist (DFR). Chips are covered/sealed by low temperature (50°C) lamination of a 50-μm-thick DFR layer having excellent optical and mechanical properties. To separate cleaned and sealed chips from the wafer, we used an effective chip singulation technique which we informally call the "chip-olate" process. In the experimental section, we first studied dielectrophoretic trapping of 10-μm beads for flow rates ranging from 80 pL s-1 to 2.5 nL s-1 that are generated by an external syringe pump. Then, we characterized the embedded microheater in DFR-covered chips. Flow rates as high as 8 nL s-1 were generated by evaporation-induced flow when the heater was biased by 10 V, corresponding to 270-mW power. Finally, DEP-based trapping and fluorescent detection of functionalized beads were demonstrated as the flow was generated by evaporation-induced flow after the microfluidic structures were filled by capillarity.

  10. Two-stage microfluidic chip for selective isolation of circulating tumor cells (CTCs).

    PubMed

    Hyun, Kyung-A; Lee, Tae Yoon; Lee, Su Hyun; Jung, Hyo-Il

    2015-05-15

    Over the past few decades, circulating tumor cells (CTCs) have been studied as a means of overcoming cancer. However, the rarity and heterogeneity of CTCs have been the most significant hurdles in CTC research. Many techniques for CTC isolation have been developed and can be classified into positive enrichment (i.e., specifically isolating target cells using cell size, surface protein expression, and so on) and negative enrichment (i.e., specifically eluting non-target cells). Positive enrichment methods lead to high purity, but could be biased by their selection criteria, while the negative enrichment methods have relatively low purity, but can isolate heterogeneous CTCs. To compensate for the known disadvantages of the positive and negative enrichments, in this study we introduced a two-stage microfluidic chip. The first stage involves a microfluidic magnetic activated cell sorting (μ-MACS) chip to elute white blood cells (WBCs). The second stage involves a geometrically activated surface interaction (GASI) chip for the selective isolation of CTCs. We observed up to 763-fold enrichment in cancer cells spiked into 5 mL of blood sample using the μ-MACS chip at 400 μL/min flow rate. Cancer cells were successfully separated with separation efficiencies ranging from 10.19% to 22.91% based on their EpCAM or HER2 surface protein expression using the GASI chip at a 100 μL/min flow rate. Our two-stage microfluidic chips not only isolated CTCs from blood cells, but also classified heterogeneous CTCs based on their characteristics. Therefore, our chips can contribute to research on CTC heterogeneity of CTCs, and, by extension, personalized cancer treatment.

  11. 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.

  12. Identification of microfluidic two-phase flow patterns in lab-on-chip devices.

    PubMed

    Yang, Zhaochu; Dong, Tao; Halvorsen, Einar

    2014-01-01

    This work describes a capacitive sensor for identification of microfluidic two-phase flow in lab-on-chip devices. With interdigital electrodes and thin insulation layer utilized, this sensor is capable of being integrated with the microsystems easily. Transducing principle and design considerations are presented with respect to the microfluidic gas/liquid flow patterns. Numerical simulation results verify the operational principle. And the factors affecting the performance of the sensor are discussed. Besides, a feasible process flow for the fabrication is also proposed.

  13. Optical fiber LPG biosensor integrated microfluidic chip for ultrasensitive glucose detection

    PubMed Central

    Yin, Ming-jie; Huang, Bobo; Gao, Shaorui; Zhang, A. Ping; Ye, Xuesong

    2016-01-01

    An optical fiber sensor integrated microfluidic chip is presented for ultrasensitive detection of glucose. A long-period grating (LPG) inscribed in a small-diameter single-mode fiber (SDSMF) is employed as an optical refractive-index (RI) sensor. With the layer-by-layer (LbL) self-assembly technique, poly (ethylenimine) (PEI) and poly (acrylic acid) (PAA) multilayer film is deposited on the SDSMF-LPG sensor for both supporting and signal enhancement, and then a glucose oxidase (GOD) layer is immobilized on the outer layer for glucose sensing. A microfluidic chip for glucose detection is fabricated after embedding the SDSMF-LPG biosensor into the microchannel of the chip. Experimental results reveal that the SDSMF-LPG biosensor based on such a hybrid sensing film can ultrasensitively detect glucose concentration as low as 1 nM. After integration into the microfluidic chip, the detection range of the sensor is extended from 2 µM to 10 µM, and the response time is remarkablely shortened from 6 minutes to 70 seconds. PMID:27231643

  14. Microfluidic chip-based technologies: emerging platforms for cancer diagnosis

    PubMed Central

    2013-01-01

    The development of early and personalized diagnostic protocols is considered the most promising avenue to decrease mortality from cancer and improve outcome. The emerging microfluidic-based analyzing platforms hold high promises to fulfill high-throughput and high-precision screening with reduced equipment cost and low analysis time, as compared to traditional bulky counterparts in bench-top laboratories. This article overviewed the potential applications of microfluidic technologies for detection and monitoring of cancer through nucleic acid and protein biomarker analysis. The implications of the technologies in cancer cytology that can provide functional personalized diagnosis were highlighted. Finally, the future niches for using microfluidic-based systems in tumor screening were briefly discussed. PMID:24070124

  15. Microfluidic organ-on-chip technology for blood-brain barrier research.

    PubMed

    van der Helm, Marinke W; van der Meer, Andries D; Eijkel, Jan C T; van den Berg, Albert; Segerink, Loes I

    2016-01-01

    Organs-on-chips are a new class of microengineered laboratory models that combine several of the advantages of current in vivo and in vitro models. In this review, we summarize the advances that have been made in the development of organ-on-chip models of the blood-brain barrier (BBBs-on-chips) and the challenges that are still ahead. The BBB is formed by specialized endothelial cells and separates blood from brain tissue. It protects the brain from harmful compounds from the blood and provides homeostasis for optimal neuronal function [corrected]. Studying BBB function and dysfunction is important for drug development and biomedical research. Microfluidic BBBs-on-chips enable real-time study of (human) cells in an engineered physiological microenvironment, for example incorporating small geometries and fluid flow as well as sensors. Examples of BBBs-on-chips in literature already show the potential of more realistic microenvironments and the study of organ-level functions. A key challenge in the field of BBB-on-chip development is the current lack of standardized quantification of parameters such as barrier permeability and shear stress. This limits the potential for direct comparison of the performance of different BBB-on-chip models to each other and existing models. We give recommendations for further standardization in model characterization and conclude that the rapidly emerging field of BBB-on-chip models holds great promise for further studies in BBB biology and drug development.

  16. Microfluidic organ-on-chip technology for blood-brain barrier research

    PubMed Central

    van der Helm, Marinke W; van der Meer, Andries D; Eijkel, Jan C T; van den Berg, Albert; Segerink, Loes I

    2016-01-01

    ABSTRACT Organs-on-chips are a new class of microengineered laboratory models that combine several of the advantages of current in vivo and in vitro models. In this review, we summarize the advances that have been made in the development of organ-on-chip models of the blood-brain barrier (BBBs-on-chips) and the challenges that are still ahead. The BBB is formed by specialized e3ndothelial cells and separates blood from brain tissue. It protects the brain from harmful compounds from the blood and provides homeostasis for optimal neuronal function. Studying BBB function and dysfunction is important for drug development and biomedical research. Microfluidic BBBs-on-chips enable real-time study of (human) cells in an engineered physiological microenvironment, for example incorporating small geometries and fluid flow as well as sensors. Examples of BBBs-on-chips in literature already show the potential of more realistic microenvironments and the study of organ-level functions. A key challenge in the field of BBB-on-chip development is the current lack of standardized quantification of parameters such as barrier permeability and shear stress. This limits the potential for direct comparison of the performance of different BBB-on-chip models to each other and existing models. We give recommendations for further standardization in model characterization and conclude that the rapidly emerging field of BBB-on-chip models holds great promise for further studies in BBB biology and drug development. PMID:27141422

  17. Interfacial tension based on-chip extraction of microparticles confined in microfluidic Stokes flows

    NASA Astrophysics Data System (ADS)

    Huang, Haishui; He, Xiaoming

    2014-10-01

    Microfluidics involving two immiscible fluids (oil and water) has been increasingly used to produce hydrogel microparticles with wide applications. However, it is difficult to extract the microparticles out of the microfluidic Stokes flows of oil that have a Reynolds number (the ratio of inertia to viscous force) much less than one, where the dominant viscous force tends to drive the microparticles to move together with the surrounding oil. Here, we present a passive method for extracting hydrogel microparticles in microfluidic Stokes flow from oil into aqueous extracting solution on-chip by utilizing the intrinsic interfacial tension between oil and the microparticles. We further reveal that the thickness of an "extended confining layer" of oil next to the interface between oil and aqueous extracting solution must be smaller than the radius of microparticles for effective extraction. This method uses a simple planar merging microchannel design that can be readily fabricated and further integrated into a fluidic system to extract microparticles for wide applications.

  18. Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip.

    PubMed

    Hou, Hsien-San; Chang, Hui-Fang; Cheng, Ji-Yen

    2015-12-29

    The behavior of directional cell migration under a direct current electric-field (dcEF) is referred to as electrotaxis. The significant role of physiological dcEF in guiding cell movement during embryo development, cell differentiation, and wound healing has been demonstrated in many studies. By applying microfluidic chips to an electrotaxis assay, the investigation process is shortened and experimental errors are minimized. In recent years, microfluidic devices made of polymeric substances (e.g., polymethylmethacrylate, PMMA, or acrylic) or polydimethylsiloxane (PDMS) have been widely used in studying the responses of cells to electrical stimulation. However, unlike the numerous steps required to fabricate a PDMS device, the simple and rapid construction of the acrylic microfluidic chip makes it suitable for both device prototyping and production. Yet none of the reported devices facilitate the efficient study of the simultaneous chemical and dcEF effects on cells. In this report, we describe our design and fabrication of an acrylic-based multichannel dual-electric-field (MDF) chip to investigate the concurrent effect of chemical and electrical stimulation on lung cancer cells. The MDF chip provides eight combinations of electrical/chemical stimulations in a single test. The chip not only greatly shortens the required experimental time but also increases accuracy in electrotaxis studies.

  19. Microfluidic positioning of pollen grains in lab-on-a-chip for single cell analysis.

    PubMed

    Ghanbari, Mahmood; Nezhad, Amir Sanati; Agudelo, Carlos G; Packirisamy, Muthukumaran; Geitmann, Anja

    2014-04-01

    A lab-on-a-chip device with a knot shaped microfluidic network is presented to enable trapping of single pollen grains at the entrances of a series of microchannels. This set-up serves to create identical growth conditions for serially arranged tip growing plant cells such as pollen tubes. The design consists of an inlet to introduce the pollen suspension into the chip, three outlets to evacuate excess medium or cells, a distribution chamber to guide the pollen grains toward the growth microchannels and a serial arrangement of microchannels with different geometries connected to the distribution chamber. These microchannels are to harbor the individual pollen tubes. Two different criteria were established to assess the efficiency and optimize the device: trapping probability and uniformity of fluid flow conditions within the microchannels. The performance of different geometries of the microfluidic network was numerically analyzed and experimentally tested.

  20. Microfluidic Organ/Body-on-a-Chip Devices at the Convergence of Biology and Microengineering.

    PubMed

    Perestrelo, Ana Rubina; Águas, Ana C P; Rainer, Alberto; Forte, Giancarlo

    2015-12-10

    Recent advances in biomedical technologies are mostly related to the convergence of biology with microengineering. For instance, microfluidic devices are now commonly found in most research centers, clinics and hospitals, contributing to more accurate studies and therapies as powerful tools for drug delivery, monitoring of specific analytes, and medical diagnostics. Most remarkably, integration of cellularized constructs within microengineered platforms has enabled the recapitulation of the physiological and pathological conditions of complex tissues and organs. The so-called "organ-on-a-chip" technology, which represents a new avenue in the field of advanced in vitro models, with the potential to revolutionize current approaches to drug screening and toxicology studies. This review aims to highlight recent advances of microfluidic-based devices towards a body-on-a-chip concept, exploring their technology and broad applications in the biomedical field.

  1. 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

  2. Single-use thermoplastic microfluidic burst valves enabling on-chip reagent storage.

    PubMed

    Rahmanian, Omid D; DeVoe, Don L

    2015-05-01

    A simple and reliable method for fabricating single-use normally closed burst valves in thermoplastic microfluidic devices is presented, using a process flow that is readily integrated into established workflows for the fabrication of thermoplastic microfluidics. An experimental study of valve performance reveals the relationships between valve geometry and burst pressure. The technology is demonstrated in a device employing multiple valves engineered to actuate at different inlet pressures that can be generated using integrated screw pumps. On-chip storage and reconstitution of fluorescein salt sealed within defined reagent chambers are demonstrated. By taking advantage of the low gas and water permeability of cyclic olefin copolymer, the robust burst valves allow on-chip hermetic storage of reagents, making the technology well suited for the development of integrated and disposable assays for use at the point of care.

  3. 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.

  4. Validation of a fully autonomous phosphate analyser based on a microfluidic lab-on-a-chip.

    PubMed

    Slater, C; Cleary, J; Lau, K-T; Snakenborg, D; Corcoran, B; Kutter, J P; Diamond, D

    2010-01-01

    This work describes the design of a phosphate analyser that utilises a microfluidic lab-on-a-chip. The analyser contains all the required chemical storage, pumping and electronic components to carry out a complete phosphate assay. The system is self-calibrating and self-cleaning, thus capable of long-term operation. This was proven by a bench top calibration of the analyser using standard solutions and also by comparing the analyser's performance to a commercially available phosphate monitor installed at a waste water treatment plant. The output of the microfluidic lab-on-a-chip analyser was shown to have sensitivity and linear range equivalent to the commercially available monitor and also the ability to operate over an extended period of time.

  5. Rheology of conductive ink flow for printed electronics on a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Jang, Young-Sik; Song, Simon

    2012-07-01

    Printed electronics have recently attracted extensive attention due to their superior productivity to conventional semiconductor fabrication methods. To develop printing devices optimized for printed electronics, numerical studies on ink flows are often necessary, and, therefore, it is critical to provide accurate ink properties for reliable numerical results. However, it is difficult to find such data in literature since inks for printed electronics contains conductive metallic nanoparticles and they are not only non-Newtonian but expensive. Thus, we propose utilizing a microfluidic chip to investigate rheological properties of conductive inks. By using micro particle image velocimeter along with an immersion oil technique, we examine the flow characteristics of two commercial conductive inks containing Ag nanoparticles on microfluidic chips. We found that the ink flows show a stronger shear-thinning behavior as the Ag content increases. Finally, suitable rheological models applicable to numerical simulations for those inks are suggested after comparing the experimental data to frequently used rheological models.

  6. Capillary-driven multiparametric microfluidic chips for one-step immunoassays.

    PubMed

    Gervais, Luc; Hitzbleck, Martina; Delamarche, Emmanuel

    2011-09-15

    Here we present a capillary-driven microfluidic chip for "one-step" immunoassays. The chip allows for easy modification of several assay parameters such as the flow rates of sample, the volumes of samples for tests, and the type of reagents and receptors for detecting analytes. We therefore term such a chip a multiparametric chip and illustrate this concept with the integration and release of anti-C-reactive protein (CRP) detection antibodies (dAbs) together with splitting flow of samples containing CRP across lines of anti-CRP capture antibodies (cAbs). The microfluidic chip is fabricated in Si and is sealed with polydimethylsiloxane (PDMS) patterned with cAbs. The microfluidic chip is ∼1.7×3.4 cm(2) and is capable of analyzing 20 μL of human serum in 6 parallel flow paths with a range of flow rates from 3.3 nL s(-1) to 0.46 nL s(-1). An inkjet spotter was used to deposit 10.6 nL of dAb solution in a structure vicinal to the main flow path of the chip. The consequent asymmetric release of dAbs in a stream of human serum is compensated by a Dean flow mixer having 9 mixing loops and a footprint of 2.8 mm × 0.78 mm. The quantity of dAb present in the half of the flow path close to the spotting region decreases from 83% at the entrance of the mixer to 52% in the region after the mixer. The sample is then equally split into 6 reaction chambers and proceeds via connecting channels to 2 μL capillary pumps. The hydraulic resistance of the connecting channels is designed to vary flow rates, and therefore the kinetics of capture of CRP-dAb complexes, from 10 min to 72 min. The increased incubation time leads to a fourfold increase in detection signal in the reaction chamber with the longer incubation time. The concept presented here is flexible and suited for implementing various surface fluorescence immunoassays on a capillary-driven microfluidic chip.

  7. A disposable microfluidic biochip with on-chip molecularly imprinted biosensors for optical detection of anesthetic propofol.

    PubMed

    Hong, Chien-Chong; Chang, Po-Hsiang; Lin, Chih-Chung; Hong, Chian-Lang

    2010-05-15

    This paper presents a disposable microfluidic biochip with on-chip molecularly imprinted biosensors for optical detection of anesthetic propofol. So far, the methods to detect anesthetic propofol in hospitals are liquid chromatography (LC), high-performance liquid chromatography (HPLC), and gas chromatography-mass spectroscopy (GC-MS). These conventional instruments are bulky, expensive, and not ease of access. In this work, a novel plastic microfluidic biochip with on-chip anesthetic biosensor has been developed and characterized for rapid detection of anesthetic propofol. The template-molecule imprinted polymers were integrated into microfluidic biochips to be used for detecting anesthetic propofol optically at 655 nm wavelength after the reaction of propofol with color reagent. Experimental results show that the sensitivity of the microfluidic biochip with on-chip molecularly imprinted polymers (MIPs) biosensor is 6.47 mV/(ppm mm(2)). The specific binding of MIP to non-imprinted polymer (NIP) is up to 456%. And the detection limit of the microsystem is 0.25 ppm with a linear detection range from 0.25 to 10 ppm. The disposable microfluidic biochip with on-chip anesthetic biosensor using molecularly imprinted polymers presented in this work showed excellent performance in separation and sensing of anesthetic propofol molecules. While compared to large-scale conventional instruments, the developed microfluidic biochips with on-chip MIP biosensors have the advantages of compact size, high sensitivity, high selectivity, low cost, and fast response.

  8. Fabrication of Poly(methyl Methacrylate) microfluidic chips by redox-initiated polymerization

    SciTech Connect

    Chen, Jiang; Lin, Yuehe; Chen, Gang

    2007-08-16

    In this report, a method based on the redox-initiated polymerization of methyl methacrylate (MMA) has been developed for the rapid fabrication of PMMA microfluidic chips.The new fabrication approach obviates the need for special equipment and significantly simplifies the process of fabricating microdevices. The attractive performance of the novel PMMA microchips has been demonstrated in connection with contactless conductivity detection for the separation and detection of ionic species.

  9. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems

    NASA Astrophysics Data System (ADS)

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-01

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems.

  10. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems.

    PubMed

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-29

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems.

  11. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems

    PubMed Central

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-01

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems. PMID:26823292

  12. A review of digital microfluidics as portable platforms for lab-on a-chip applications.

    PubMed

    Samiei, Ehsan; Tabrizian, Maryam; Hoorfar, Mina

    2016-07-07

    Following the development of microfluidic systems, there has been a high tendency towards developing lab-on-a-chip devices for biochemical applications. A great deal of effort has been devoted to improve and advance these devices with the goal of performing complete sets of biochemical assays on the device and possibly developing portable platforms for point of care applications. Among the different microfluidic systems used for such a purpose, digital microfluidics (DMF) shows high flexibility and capability of performing multiplex and parallel biochemical operations, and hence, has been considered as a suitable candidate for lab-on-a-chip applications. In this review, we discuss the most recent advances in the DMF platforms, and evaluate the feasibility of developing multifunctional packages for performing complete sets of processes of biochemical assays, particularly for point-of-care applications. The progress in the development of DMF systems is reviewed from eight different aspects, including device fabrication, basic fluidic operations, automation, manipulation of biological samples, advanced operations, detection, biological applications, and finally, packaging and portability of the DMF devices. Success in developing the lab-on-a-chip DMF devices will be concluded based on the advances achieved in each of these aspects.

  13. 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.

  14. Refolding of difficult-to-fold proteins by a gradual decrease of denaturant using microfluidic chips.

    PubMed

    Yamaguchi, Hiroshi; Miyazaki, Masaya; Briones-Nagata, Maria Portia; Maeda, Hideaki

    2010-06-01

    Protein refolding is an important process to obtain active recombinant proteins from inclusion bodies (protein aggregates). However, the conventional refolding method of dialysis or dilution is a time consuming procedure and often, recovering yields of active proteins are low. In this study, we used controllable diffusion through laminar flow in microchannels to control the denaturant concentration. The performance of the designed microfluidic chips was evaluated by the refolding of difficult-to-fold proteins (citrate synthase and the zeta-associated protein 70-kDa protein kinase domain). We demonstrated this by varying the flow rates of the diluting buffer stream(s) and multi-junctions which could control the different flow rate ratios of the buffer stream(s) and the denatured protein stream. By this strategy, we were able to improve the efficiency of protein refolding. Our method achieved refolding within a short period of time at room temperature without the need of any small molecules or chaperone proteins. Moreover, the efficiency of protein refolding by microfluidic chip was found higher than that prepared by dialysis or dilution. These results suggest that microfluidic chips employing this strategy may provide miniaturized tools for rapid and efficient recovery of active proteins from inclusion bodies.

  15. Recent Results of the Investigation of a Microfluidic Sampling Chip and Sampling System for Hot Cell Aqueous Processing Streams

    SciTech Connect

    Julia Tripp; Jack Law; Tara Smith

    2013-10-01

    A Fuel Cycle Research and Development project has investigated an innovative sampling method that could evolve into the next generation sampling and analysis system for metallic elements present in aqueous processing streams. Initially sampling technologies were evaluated and microfluidics sampling chip technology was selected and tested. A conceptual design for a fully automated microcapillary-based system was completed and a robotic automated sampling system was fabricated. The mechanical and sampling operation of the completed sampling system was investigated. In addition, the production of a less expensive, mass produced sampling chip was investigated to avoid chip reuse thus increasing sampling reproducibility/accuracy. The microfluidic-based robotic sampling system’s mechanical elements were tested to ensure analytical reproducibility and the optimum robotic handling of microfluidic sampling chips.

  16. Scattering detection using a photonic-microfluidic integrated device with on-chip collection capabilities.

    PubMed

    Watts, Benjamin R; Zhang, Zhiyi; Xu, Chang Qing; Cao, Xudong; Lin, Min

    2014-02-01

    SU-8-based photonic-microfluidic integrated devices with on-chip beam shaping and collection capabilities were demonstrated in a scattering detection and counting application. Through the proper deployment of the tailored beam geometries via the on-chip excitation optics, excellent CV values were measured for 1, 2, and 5 μm blank beads, 16.4, 11.0, and 12.5%, respectively, coupled with a simple free-space optical detection scheme. The performance of these devices was found dependent on the combination of on-chip, lens-shaped beam geometry and bead size. While very low CVs were obtained when the combination was ideal, a nonideal combination could still result in acceptable CVs for flow cytometry; the reliability was confirmed via devices being able to resolve separate populations of 2.0 and 5.0 μm beads from their mixture with low CV values of 15.9 and 18.5%, respectively. On-chip collection using integrated on-chip optical waveguides was shown to be very reliable in comparison with a free-space collection scheme, yielding a coincident rate of 94.2%. A CV as low as 19.2% was obtained from the on-chip excitation and collection of 5 μm beads when the on-chip lens-shaped beam had a 6.0-μm beam waist.

  17. Microfluidic chip-based analytical system for rapid screening of photocatalysts.

    PubMed

    Zhang, Hao; Wang, Jing-Jing; Fan, Jie; Fang, Qun

    2013-11-15

    A simple and efficient microfluidic chip-based analytical system for rapid screening of photocatalysts was developed. The catalyst screening system consisted of a microchip with multiple channels for parallel reactions, a UV light source, and a CCD camera-based photometric detection system for monitoring the photocatalytic reaction. A novel microfluidic introduction method for loading particle samples into chip microchannels was established using dry sample powders and wedge-structure channel design. With this method, multiple different photocatalyst samples could be quickly introduced into the microchip with good reproducibility without the need of additional pumps or valves. We applied the present system in the rapid screening of doping TiO2 photocatalysts in terms of their activity for methylene blue (MB) degradation under UV light irradiation. Ten parallel photocatalyst screening reactions were achieved within 15 min in the multi-channel chip. We also examined nine element doped TiO2 materials to investigate the doping effects of different elements on TiO2. Compared with conventional systems, the photocatalyst consumption (0.1mg) in the present system was significantly reduced at least 100 times. High reaction rate in chip microreactors was obtained with an increase of two orders of magnitude over bulk reactors. The miniaturization of the photocatalytic reaction on the microchip significantly improves the reaction rates, reduces the sample and reagent consumptions, and increases the throughput of screening for multiple catalyst samples in parallel. The present work provides a novel application for microfluidic chip-based analytical systems, as well as a rapid, highly-efficient and low-consumption method for screening of photocatalysts.

  18. Capillary-driven microfluidic chips with evaporation-induced flow control and dielectrophoretic microbead trapping

    NASA Astrophysics Data System (ADS)

    Temiz, Yuksel; Skorucak, Jelena; Delamarche, Emmanuel

    2014-03-01

    This work reports our efforts on developing simple-to-use microfluidic devices for point-of-care diagnostic applications with recent extensions that include the trapping of microbeads using dielectrophoresis (DEP) and the modulation of capillary-driven flow using integrated microheaters. DEP serves the purpose of trapping microbeads coated with receptors and analytes for detection of a fluorescent signal. The microheater is actuated once the chip is filled by capillarity, creating an evaporation-induced flow tuned according to assay conditions. The chips are composed of a glass substrate patterned with 50-nm-thick Pd electrodes and microfluidic structures made using a 20-μm-thick dry-film resist (DFR). Chips are covered/sealed by low-temperature (50 °C) lamination of a 50-μm-thick DFR layer having excellent optical and mechanical properties. To separate cleaned and sealed chips from the wafer, we used an effective chip singulation technique that we informally call the "chip-olate" process. In the experimental section, we first studied dielectrophoretic trapping of 10 μm beads for flow rates ranging from 80 pL s-1 to 2.5 nL s-1 and that are generated by an external syringe pump. Then, we characterized the embedded microheater in DFR-covered chips. Flow rates as high as 8 nL s-1 were generated by evaporation-induced flow when the heater was biased by 10 V, corresponding to 270 mW power. Finally, DEP-based trapping and fluorescent detection of functionalized beads were demonstrated as the flow was generated by the combination of capillary filling and evaporation-induced flow.

  19. Development of a microfluidic "click chip" incorporating an immobilized Cu(I) catalyst.

    PubMed

    Li, Hairong; Whittenberg, Joseph J; Zhou, Haiying; Ranganathan, David; Desai, Amit V; Koziol, Jan; Zeng, Dexing; Kenis, Paul J A; Reichert, David E

    2015-01-01

    We have developed a microfluidic "click chip" incorporating an immobilized Cu(I) catalyst for click reactions. The microfluidic device was fabricated from polydimethylsiloxane (PDMS) bonded to glass and featured ~14,400 posts on the surface to improve catalyst immobilization. This design increased the immobilization efficiency and reduces the reagents' diffusion time to active catalyst site. The device also incorporates five reservoirs to increase the reaction volume with minimal hydrodynamic pressure drop across the device. A novel water-soluble tris-(benzyltriazolylmethyl)amine (TBTA) derivative capable of stabilizing Cu(I), ligand 2, was synthesized and successfully immobilized on the chip surface. The catalyst immobilized chip surface was characterized by X-ray photoelectron spectroscopy (XPS). The immobilization efficiency was evaluated via radiotracer methods: the immobilized Cu(I) was measured as 1136±272 nmol and the surface immobilized Cu(I) density was 81±20 nmol cm(-2). The active Cu(I)-ligand 2 could be regenerated up to five times without losing any catalyst efficiency. The "click" reaction of Flu568-azide and propargylamine was studied on chip for proof-of-principle. The on-chip reaction yields were ca. 82% with a 50 min reaction time or ca. 55% with a 15 min period at 37 °C, which was higher than those obtained in the conventional reaction. The on-chip "click" reaction involving a biomolecule, cyclo(RGDfK) peptide was also studied and demonstrated a conversion yield of ca. 98%. These encouraging results show promise on the application of the Cu(I) catalyst immobilized "click chip" for the development of biomolecule based imaging agents.

  20. Single-layer planar on-chip flow cytometer using microfluidic drifting based three-dimensional (3D) hydrodynamic focusing.

    PubMed

    Mao, Xiaole; Lin, Sz-Chin Steven; Dong, Cheng; Huang, Tony Jun

    2009-06-07

    In this work, we demonstrate an on-chip microfluidic flow cytometry system based on a three-dimensional (3D) hydrodynamic focusing technique, microfluidic drifting. By inducing Dean flow in a curved microfluidic channel, microfluidic drifting can be used to hydrodynamically focus cells or particles in the vertical direction and enables the 3D hydrodynamic focusing in a single-layer planar microfluidic device. Through theoretical calculation, numerical simulation, and experimental characterization, we found that the microfluidic drifting technique can be effectively applied to three-dimensionally focus microparticles with density and size equivalent to those of human CD4+ T lymphocytes. In addition, we developed a flow cytometry platform by integrating the 3D focusing device with a laser-induced fluorescence (LIF) detection system. The system was shown to provide effective high-throughput flow cytometry measurements at a rate of greater than 1700 cells s(-1).

  1. Temperature regulation during ultrasonic manipulation for long-term cell handling in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Svennebring, J.; Manneberg, O.; Wiklund, M.

    2007-12-01

    We demonstrate simultaneous micromanipulation and temperature regulation by the use of ultrasonic standing wave technology in a microfluidic chip. The system is based on a microfabricated silicon structure sandwiched between two glass layers, and an external ultrasonic transducer using a refractive wedge placed on top of the chip for efficient coupling of ultrasound into the microchannel. The chip is fully transparent and compatible with any kind of high-resolution optical microscopy. The temperature regulation method uses calibration data of the temperature increase due to the ultrasonic actuation for determining the temperature of the surrounding air and microscope table, controlled by a warm-air heating unit and a heatable mounting frame. The heating methods are independent of each other, resulting in a flexible choice of ultrasonic actuation voltage and flow rate for different cell and particle manipulation purposes. Our results indicate that it is possible to perform stable temperature regulation with an accuracy of the order of ±0.1 °C around any physiologically relevant temperature (e.g., 37 °C) with high temporal stability and repeatability. The purpose is to use ultrasound for long-term cell and/or particle handling in a microfluidic chip while controlling and maintaining the biocompatibility of the system.

  2. Integration of programmable microfluidics and on-chip fluorescence detection for biosensing applications

    PubMed Central

    Parks, J. W.; Olson, M. A.; Kim, J.; Ozcelik, D.; Cai, H.; Carrion, R.; Patterson, J. L.; Mathies, R. A.; Hawkins, A. R.; Schmidt, H.

    2014-01-01

    We describe the integration of an actively controlled programmable microfluidic sample processor with on-chip optical fluorescence detection to create a single, hybrid sensor system. An array of lifting gate microvalves (automaton) is fabricated with soft lithography, which is reconfigurably joined to a liquid-core, anti-resonant reflecting optical waveguide (ARROW) silicon chip fabricated with conventional microfabrication. In the automaton, various sample handling steps such as mixing, transporting, splitting, isolating, and storing are achieved rapidly and precisely to detect viral nucleic acid targets, while the optofluidic chip provides single particle detection sensitivity using integrated optics. Specifically, an assay for detection of viral nucleic acid targets is implemented. Labeled target nucleic acids are first captured and isolated on magnetic microbeads in the automaton, followed by optical detection of single beads on the ARROW chip. The combination of automated microfluidic sample preparation and highly sensitive optical detection opens possibilities for portable instruments for point-of-use analysis of minute, low concentration biological samples. PMID:25584111

  3. 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

  4. Asymmetric cancer-cell filopodium growth induced by electric-fields in a microfluidic culture chip.

    PubMed

    Wang, Chun-Chieh; Kao, Yu-Chiu; Chi, Pei-Yin; Huang, Ching-Wen; Lin, Jiunn-Yuan; Chou, Chia-Fu; Cheng, Ji-Yen; Lee, Chau-Hwang

    2011-02-21

    We combine a micro-fluidic electric-field cell-culture (MEC) chip with structured-illumination nano-profilometry (SINAP) to quantitatively study the variations of cancer cell filopodia under external direct-current electric field (dcEF) stimulations. Because the lateral resolution of SINAP is better than 150 nm in bright-field image modality, filopodia with diameters smaller than 200 nm can be observed clearly without fluorescent labeling. In the MEC chip, a homogeneous EF is generated inside the culture area that simulates the endogenous EF environment. With this MEC chip-SINAP system, we directly observe and quantify the biased growth of filopodia of lung cancer cells toward the cathode. The epidermal growth factor receptors around the cell edges are also redistributed to the cathodal side. These results suggest that cancer-cell filopodia respond to the changes in EFs in the microenvironment.

  5. A chitosan coated monolith for nucleic acid capture in a thermoplastic microfluidic chip

    PubMed Central

    Kendall, Eric L.; Wienhold, Erik; DeVoe, Don L.

    2014-01-01

    A technique for microfluidic, pH modulated DNA capture and purification using chitosan functionalized glycidyl methacrylate monoliths is presented. Highly porous polymer monoliths are formed and subsequently functionalized off-chip in a batch process before insertion into thermoplastic microchannels prior to solvent bonding, simplifying the overall fabrication process by eliminating the need for on-chip surface modifications. The monolith anchoring method allows for the use of large cross-section monoliths enabling high flowrates and high DNA capture capacity with a minimum of added design complexity. Using monolith capture elements requiring less than 1 mm2 of chip surface area, loading levels above 100 ng are demonstrated, with DNA capture and elution efficiency of 54.2% ± 14.2% achieved. PMID:25379094

  6. Microfluidic chip for stacking, separation and extraction of multiple DNA fragments.

    PubMed

    Wu, Ruige; Seah, Y P; Wang, Zhiping

    2016-03-11

    A disposable integrated microfluidic device was developed for rapid sample stacking, separation and extraction of multiple DNA fragments from a relatively large amount of sample. Isotachophoresis hyphenated gel electrophoresis (ITP-GE) was used to pre-concentrate and separate DNA fragments, followed by extraction of pure DNA fragments with electroelution on-chip. DNA fragments of 200bp, 500bp and 1kbp were successfully separated and collected in the extraction chamber within 25min. The extraction efficiency obtained from the chip was 49.9%, 52.1% and 53.7% for 200bp, 500bp and 1kbp DNA fragments, respectively. The extracted DNA fragments exhibited compatibility with downstream enzymatic reactions, for example PCR. The chip was also used to extract DNA fragments with specific size range from sheared genomic DNA and demonstrated similar performance to that using traditional gel cutting method. The whole assay can finish in 32min, 6 times faster than traditional method.

  7. Room-temperature bonding for plastic high-pressure microfluidic chips.

    PubMed

    Mair, Dieudonne A; Rolandi, Marco; Snauko, Marian; Noroski, Richard; Svec, Frantisek; Fréchet, Jean M J

    2007-07-01

    A generic method for the rapid, reproducible, and robust bonding of microfluidic chips fabricated from plastics has been developed and optimized. One of the bonding surfaces is exposed to solvent vapor prior to bringing the mating parts into contact and applying a load. Nanoindentation measurements performed by atomic force microscopy show that a reversible material softening occurs upon exposure to solvent vapor. Subsequent exposure of the bonded chip to UV light then strengthens the bond between mating parts and increases the burst pressure by 50% due to partial cross-linking and chain scission reactions as measured by size exclusion chromatography-multiangle light scattering (SEC-MALS). Performing all steps of this procedure at room temperature eliminates channel distortion observed during thermal bonding and affords channels with highly uniform cross-sectional dimensions. Our technique enables chips resistant to pressures as high as 34.6 MPa.

  8. Rapid cell-patterning and microfluidic chip fabrication by crack-free CO2 laser ablation on glass

    NASA Astrophysics Data System (ADS)

    Yen, Meng-Hua; Cheng, Ji-Yen; Wei, Cheng-Wey; Chuang, Yung-Chuan; Young, Tai-Horng

    2006-07-01

    This paper uses a widely available CO2 laser scriber (λ = 10.6 µm) to perform the direct-writing ablation of quartz, borofloat and pyrex substrates for the development of microfluidic chips and cell chips. The surface quality of the ablated microchannels and the presence of debris and distortion are examined by scanning electron microscopy, atomic force microscopy and surface profile measurement techniques. The developed laser ablation system provides a versatile and economic approach for the fabrication of glass microfluidic chips with crack-free structures. In the laser writing process, the desired microfluidic patterns are designed using commercial computer software and are then transferred to the laser scriber to ablate the trenches. This process eliminates the requirement for corrosive chemicals and photomasks, and hence the overall microchip development time is limited to less than 24 h. Additionally, since the laser writing process is not limited by the dimensions of a photomask, the microchannels can be written over a large substrate area. The machining capability and versatility of the laser writing system are demonstrated through its application to the fabrication of a borofloat microfluidic chip and the writing of a series of asymmetric trenches in a microwell array. It is shown that the minimum attainable trench width is 95 µm and that the maximum trench depth is 225 µm. The system provides an economic and powerful means of rapid glass microfluidic chip development. A rapid cell-patterning method based on this method is also demonstrated.

  9. Microfluidic chip containing porous gradient for chemotaxis study

    NASA Astrophysics Data System (ADS)

    Al-Abboodi, Aswan; Tjeung, Ricky; Doran, Pauline; Yeo, Leslie; Friend, James; Chan, Peggy

    2011-12-01

    We have developed a new porous gradient microfluidic device based on in situ Gtn-HPA/CMC-Tyr hydrogel that comprises gelatin hydroxyphenylpropionic acid (Gtn-HPA) conjugate and carboxymethyl cellulose tyramine (CMC-Tyr) conjugate. The device is fabricated using a soft lithographic technique, in which microstructures were patterned on a thin layer of polydimethylsiloxane (PDMS) using a polymeric mold. Human fibrosarcoma cells (HT1080) were employed as invasive cancer cell model. Porosity gradients were generated by flowing pore etching fluid in the gradient generator network. Results suggested that spatial control of the porosity can be obtained, which mimics the 3-dimensional microenvironment in vivo for cell-based screening applications including real time chemotaxis, cytotoxicity, and continuous drug-response monitoring. A chemoattractant gradient is then generated and cell migration is monitored in real time using fluorescence microscopy. The viability of cells was evaluated using calcien AM stain. Herein, we successfully monitored the chemotactic responses of cancer cells, confirmed the validity of using in situ porous hydrogels as a construction material for a microchemotaxis device, and demonstrated the potential of the hydrogel with tunable porosity based microfluidic device in biological experiments. This device will also be practical in controlling the chemical and mechanical properties of the surroundings during the formation of tissue engineered constructs.

  10. A microfluidic chip for controlled release of drugs from microcapsules

    PubMed Central

    Cheng, Wen-Chuan; He, Yuan; Chang, An-Yi; Que, Long

    2013-01-01

    A new microfluidic device with liquid-droplet merging and droplet storage functions for the controlled release of drugs from microcapsules is reported. A switching channel is designed and integrated within the microfluidic device, facilitating the generation and capturing of uniform droplets by the storage chambers. The drug model is the MnCO3 microparticle, which is encapsulated by a microcapsule and fabricated using a simple layer-by-layer nanoassembly process. The merging function is used for dynamically adding the control solution into the droplets, which contain drugs within the microcapsules (DWμCs) and water. The storage chambers are used for collecting DWμCs-laden droplets so that the controlled-drug release in specific droplets can be monitored for an extended period of time, which has been experimentally implemented successfully. This technology could offer a promising technical platform for the long-term observation and studies of drug effects on specific cells in a controlled manner, which is especially useful for single cell analysis. PMID:24396536

  11. On-chip integration of droplet microfluidics and nanostructure-initiator mass spectrometry for enzyme screening.

    PubMed

    Heinemann, Joshua; Deng, Kai; Shih, Steve C C; Gao, Jian; Adams, Paul D; Singh, Anup K; Northen, Trent R

    2017-01-17

    Biological assays often require expensive reagents and tedious manipulations. These shortcomings can be overcome using digitally operated microfluidic devices that require reduced sample volumes to automate assays. One particular challenge is integrating bioassays with mass spectrometry based analysis. Towards this goal we have developed μNIMS, a highly sensitive and high throughput technique that integrates droplet microfluidics with nanostructure-initiator mass spectrometry (NIMS). Enzyme reactions are carried out in droplets that can be arrayed on discrete NIMS elements at defined time intervals for subsequent mass spectrometry analysis, enabling time resolved enzyme activity assay. We apply the μNIMS platform for kinetic characterization of a glycoside hydrolase enzyme (CelE-CMB3A), a chimeric enzyme capable of deconstructing plant hemicellulose into monosaccharides for subsequent conversion to biofuel. This study reveals NIMS nanostructures can be fabricated into arrays for microfluidic droplet deposition, NIMS is compatible with droplet and digital microfluidics, and can be used on-chip to assay glycoside hydrolase enzyme in vitro.

  12. High adhesion strength and hybrid irreversible/reversible full-PDMS microfluidic chips.

    PubMed

    Shiroma, Letícia S; Oliveira, Aline F; Lobo-Júnior, Eulicio O; Coltro, Wendell K T; Gobbi, Angelo L; de La Torre, Lucimara G; Lima, Renato S

    2017-01-25

    To the best of our knowledge, this paper outlines for the first time high adhesion and hybrid irreversible/reversible microfluidic devices fully composed of polydimethylsiloxane (PDMS). These chips were fabricated by the sandwich bonding (SWB), a method that was recently deployed by our group. SWB offers simple, fast, and low cost operation requiring only a laboratory oven. The devices showed burst pressures of up to 4.5 MPa. This value is more than tenfold the pressures withstood by the full-PDMS chips described in literature. In terms of the reversible behavior, the ability for disassembling the chip slides is crucial in research and development stages, especially when the device integrates high-cost components or harsh cleaning steps are needed. Following successive steps of detachment and bonding, the channels still withstood high pressures of approximately 1.8 MPa. Finally, the emulsification of corn oil 4.0% w/w to polyglycerol polyricinoleate with 10.0 μmol L(-1) rhodamine B aqueous solution was realized to show the relevance in enhancing the flow rate in microfluidics. Such experiment was conducted at total flow rates of 0.8-160.0 μL min(-1). The decrease in size and polydispersity of the droplets was observed at increasing flow rates. Monodisperse emulsions were achieved only at 160.0 μL min(-1).

  13. Microfluidic chip integrating high throughput continuous-flow PCR and DNA hybridization for bacteria analysis.

    PubMed

    Jiang, Xiran; Shao, Ning; Jing, Wenwen; Tao, Shengce; Liu, Sixiu; Sui, Guodong

    2014-05-01

    Rapid identification of clinical pathogens is the initial and essential step for antimicrobial therapy. Herein, we successfully developed a microfluidic device which combines high-throughput continuous-flow PCR and DNA hybridization for the detection of various bacterial pathogens. Universal primers were designed based on the conserved regions of bacterial 16S ribosomal DNA (16S rDNA), and specific probes were designed from a variable region of 16S rDNA within the amplicon sequences. In the chip operation, after the continuous flow PCR was achieved in the first microfluidic chip, the product was directly introduced into a hybridization chip integrated with microarray containing the immobilized DNA probes. The target-probe hybridization was completed within 1h at 55 °C, and fluorescence signals were obtained as the readout. The presented device is simple, versatile and with less sample consumption compared with traditional instruments. It can perform high-throughput bacteria detections continuously in a single assay, which makes it a promising platform for clinical bacteria identifications.

  14. Fabrication of anti-protein-fouling poly(ethylene glycol) microfluidic chip electrophoresis by sandwich photolithography.

    PubMed

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Liu, Huwei; Yuan, Hua

    2016-07-01

    Microfluidic chip electrophoresis (MCE) is a powerful separation tool for biomacromolecule analysis. However, adsorption of biomacromolecules, particularly proteins onto microfluidic channels severely degrades the separation performance of MCE. In this paper, an anti-protein-fouling MCE was fabricated using a novel sandwich photolithography of poly(ethylene glycol) (PEG) prepolymers. Photopatterned microchannel with a minimum resolution of 10 μm was achieved. After equipped with a conventional online electrochemical detector, the device enabled baseline separation of bovine serum albumin, lysozyme (Lys), and cytochrome c (Cyt-c) in 53 s under a voltage of 200 V. Compared with a traditional polydimethylsiloxane MCE made by soft lithography, the PEG MCE made by the sandwich photolithography not only eliminated the need of a master mold and the additional modification process of the microchannel but also showed excellent anti-protein-fouling properties for protein separation.

  15. Study of individual erythrocyte deformability susceptibility to INFeD and ethanol using a microfluidic chip

    PubMed Central

    Liu, Lihong; Huang, Sha; Xu, Xiaoying; Han, Jongyoon

    2016-01-01

    Human red blood cells (RBCs) deformability in vitro was assessed during iron dextran (INFeD) loading and/or ethanol co-administration using microfluidic deformability screening. The results showed donor-specific variations in dose dependent deformability shift were revealed below 500 μg/mL iron dextran. Two out of nine blood samples exhibited significant cell stiffening at 500 μg/mL iron dextran loading concentration (p < 0.05, Tukey test). More interestingly, co-administration of moderate amount of ethanol was identified to have significant protective effects on RBC deformability. We also noted that ethanol can reverse the deformability of impaired RBCs. Meanwhile obvious donor dependent response to ethanol administration on RBC deformability was noted using our biomimetic microfluidic chip. PMID:26964754

  16. Heteronanojunctions with atomic size control using a lab-on-chip electrochemical approach with integrated microfluidics.

    PubMed

    Lunca Popa, P; Dalmas, G; Faramarzi, V; Dayen, J F; Majjad, H; Kemp, N T; Doudin, B

    2011-05-27

    A versatile tool for electrochemical fabrication of heteronanojunctions with nanocontacts made of a few atoms and nanogaps of molecular spacing is presented. By integrating microfluidic circuitry in a lab-on-chip approach, we keep control of the electrochemical environment in the vicinity of the nanojunction and add new versatility for exchanging and controlling the junction's medium. Nanocontacts made of various materials by successive local controlled depositions are demonstrated, with electrical properties revealing sizes reaching a few atoms only. Investigations on benchmark molecular electronics material, trapped between electrodes, reveal the possibility to create nanogaps of size matching those of molecules. We illustrate the interest of a microfluidic approach by showing that exposure of a fabricated molecular junction to controlled high solvent flows can be used as a reliability criterion for the presence of molecular entities in a gap.

  17. Heteronanojunctions with atomic size control using a lab-on-chip electrochemical approach with integrated microfluidics

    NASA Astrophysics Data System (ADS)

    Lunca Popa, P.; Dalmas, G.; Faramarzi, V.; Dayen, J. F.; Majjad, H.; Kemp, N. T.; Doudin, B.

    2011-05-01

    A versatile tool for electrochemical fabrication of heteronanojunctions with nanocontacts made of a few atoms and nanogaps of molecular spacing is presented. By integrating microfluidic circuitry in a lab-on-chip approach, we keep control of the electrochemical environment in the vicinity of the nanojunction and add new versatility for exchanging and controlling the junction's medium. Nanocontacts made of various materials by successive local controlled depositions are demonstrated, with electrical properties revealing sizes reaching a few atoms only. Investigations on benchmark molecular electronics material, trapped between electrodes, reveal the possibility to create nanogaps of size matching those of molecules. We illustrate the interest of a microfluidic approach by showing that exposure of a fabricated molecular junction to controlled high solvent flows can be used as a reliability criterion for the presence of molecular entities in a gap.

  18. Preparation of monodisperse PEG hydrogel composite microspheres via microfluidic chip with rounded channels

    NASA Astrophysics Data System (ADS)

    Yu, Bing; Cong, Hailin; Liu, Xuesong; Ren, Yumin; Wang, Jilei; Zhang, Lixin; Tang, Jianguo; Ma, Yurong; Akasaka, Takeshi

    2013-09-01

    An effective microfluidic method to fabricate monodisperse polyethylene glycol (PEG) hydrogel composite microspheres with tunable dimensions and properties is reported in this paper. A T-junction microfluidic chip equipped with rounded channels and online photopolymerization system is applied for the microsphere microfabrication. The shape and size of the microspheres are well controlled by the rounded channels and PEG prepolymer/silicon oil flow rate ratios. The obtained PEG/aspirin composite microspheres exhibit a sustained release of aspirin for a wide time range; the obtained PEG/Fe3O4 nanocomposite microspheres exhibit excellent magnetic properties; and the obtained binary PEG/dye composite microspheres show the ability to synchronously load two functional components in the same peanut-shaped or Janus hydrogel particles.

  19. Microfluidic Organ/Body-on-a-Chip Devices at the Convergence of Biology and Microengineering

    PubMed Central

    Perestrelo, Ana Rubina; Águas, Ana C. P.; Rainer, Alberto; Forte, Giancarlo

    2015-01-01

    Recent advances in biomedical technologies are mostly related to the convergence of biology with microengineering. For instance, microfluidic devices are now commonly found in most research centers, clinics and hospitals, contributing to more accurate studies and therapies as powerful tools for drug delivery, monitoring of specific analytes, and medical diagnostics. Most remarkably, integration of cellularized constructs within microengineered platforms has enabled the recapitulation of the physiological and pathological conditions of complex tissues and organs. The so-called “organ-on-a-chip” technology, which represents a new avenue in the field of advanced in vitro models, with the potential to revolutionize current approaches to drug screening and toxicology studies. This review aims to highlight recent advances of microfluidic-based devices towards a body-on-a-chip concept, exploring their technology and broad applications in the biomedical field. PMID:26690442

  20. Determination of Apparent Amylose Content in Rice by Using Paper-Based Microfluidic Chips.

    PubMed

    Hu, Xianqiao; Lu, Lin; Fang, Changyun; Duan, Binwu; Zhu, Zhiwei

    2015-11-11

    Determination of apparent amylose content in rice is a key function for rice research and the rice industry. In this paper, a novel approach with paper-based microfluidic chip is reported to determine apparent amylose content in rice. The conventional color reaction between amylose and iodine was employed. Blue color of amylose-iodine complex generated on-chip was converted to gray and measured with Photoshop after the colored chip was scanned. The method for preparation of the paper chip is described. In situ generation of iodine for on-chip color reaction was designed, and factors influencing color reaction were investigated in detail. Elimination of yellow color interference of excess iodine by exploiting color removal function of Photoshop was presented. Under the optimized conditions, apparent amylose content in rice ranging from 1.5 to 26.4% can be determined, and precision was 6.3%. The analytical results obtained with the developed approach were in good agreement with those with the continuous flow analyzer method.

  1. Development of multistage distillation in a microfluidic chip.

    PubMed

    Lam, K F; Cao, E; Sorensen, E; Gavriilidis, A

    2011-04-07

    Although there has been a lot of work on the development of microchemical processing systems such as micro-reactors and micro-sensors, little attention has been paid to micro-separation units, and in particular, microscale distillation. In this paper, various silicon-glass microscale distillation chips with different channel configurations were fabricated and tested. A temperature gradient was setup across the chip by heating and cooling the two ends. The feed was located at the middle of the microchannel. Arrays of micropillars were incorporated in order to guide the liquid flow. It was found that the separation performance was promoted by increasing the length of the microchannel. However, this created an imbalance of the liquid flows at the two sides of the microchannel and caused flooding. This hydrodynamic limitation was addressed by incorporating micropillars on both sides of the channel. The most efficient microdistillation chip consisted of a microchannel with 600 microns width and 40 cm length. Experimental results showed high efficiency for the separation of a 50 mol% acetone-water mixture when the heating and cooling temperature were 95 °C and 42 °C respectively. The concentrations of acetone were 3 mol% in the bottom stream and 95 mol% in the distillate, which was equivalent to at least 4 equilibrium stages at total reflux conditions. Furthermore, a 50 mol% methanol-toluene mixture was separated into nearly pure toluene in the bottom stream and 75 mol% methanol in the distillate. The performance of the microdistillation unit was reproducible in repeated tests.

  2. A Novel Electrochemical Microfluidic Chip Combined with Multiple Biomarkers for Early Diagnosis of Gastric Cancer

    NASA Astrophysics Data System (ADS)

    Xie, Yao; Zhi, Xiao; Su, Haichuan; Wang, Kan; Yan, Zhen; He, Nongyue; Zhang, Jingpu; Chen, Di; Cui, Daxiang

    2015-12-01

    Early diagnosis is very important to improve the survival rate of patients with gastric cancer and to understand the biology of cancer. In order to meet the clinical demands for early diagnosis of gastric cancer, we developed a disposable easy-to-use electrochemical microfluidic chip combined with multiple antibodies against six kinds of biomarkers (carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), Helicobacter pylori CagA protein (H.P.), P53oncoprotein (P53), pepsinogen I (PG I), and PG-II). The six kinds of biomarkers related to gastric cancer can be detected sensitively and synchronously in a short time. The specially designed three electrodes system enables cross-contamination to be avoided effectively. The linear ranges of detection of the electrochemical microfluidic chip were as follows: 0.37-90 ng mL-1 for CEA, 10.75-172 U mL-1 for CA19-9, 10-160 U L-1 for H.P., 35-560 ng mL-1 for P53, 37.5-600 ng mL-1 for PG I, and 2.5-80 ng mL-1for PG II. This method owns better sensitivity compared with enzyme-linked immunosorbent assay (ELISA) results of 394 specimens of gastric cancer sera. Furthermore, we established a multi-index prediction model based on the six kinds of biomarkers for predicting risk of gastric cancer. In conclusion, the electrochemical microfluidic chip for detecting multiple biomarkers has great potential in applications such as early screening of gastric cancer patients, and therapeutic evaluation, and real-time dynamic monitoring the progress of gastric cancer in near future.

  3. Immuno-capture and in situ detection of Salmonella typhimurium on a novel microfluidic chip.

    PubMed

    Wang, Renjie; Ni, Yanan; Xu, Yi; Jiang, Yan; Dong, Chunyan; Chuan, Na

    2015-01-01

    The new method presented in this article achieved the goal of capturing Salmonella typhimurium via immunoreaction and rapid in situ detection of the CdSe/ZnS quantum dots (QDs) labeled S. typhimurium by self-assembly light-emitting diode-induced fluorescence detection (LIF) microsystem on a specially designed multichannel microfluidic chip. CdSe/ZnS QDs were used as fluorescent markers improving detection sensitivity. The microfluidic chip developed in this study was composed of 12 sample channels, 3 mixing zones, and 6 immune reaction zones, which also acted as fluorescence detection zones. QDs-IgG-primary antibody complexes were generated by mixing CdSe/ZnS QDs conjugated secondary antibody (QDs-IgG) and S. typhimurium antibody (primary antibody) in mixing zones. Then, the complexes went into immune reaction zones to label previously captured S. typhimurium in the sandwich mode. The capture rate of S. typhimurium in each detection zone was up to 70%. The enriched QDs-labeled S. typhimurium was detected using a self-assembly LIF microsystem. A good linear relationship was obtained in the range from 3.7×10 to 3.7×10(5) cfu mL(-1) using the equation I=0.1739 log (C)-0.1889 with R(2)=0.9907, and the detection limit was down to 37 cfu mL(-1). The proposed method of online immunolabeling with QDs for in situ fluorescence detection on the designed multichannel microfluidic chip had been successfully used to detect S. typhimurium in pork sample, and it has shown potential advantages in practice.

  4. A Novel Electrochemical Microfluidic Chip Combined with Multiple Biomarkers for Early Diagnosis of Gastric Cancer.

    PubMed

    Xie, Yao; Zhi, Xiao; Su, Haichuan; Wang, Kan; Yan, Zhen; He, Nongyue; Zhang, Jingpu; Chen, Di; Cui, Daxiang

    2015-12-01

    Early diagnosis is very important to improve the survival rate of patients with gastric cancer and to understand the biology of cancer. In order to meet the clinical demands for early diagnosis of gastric cancer, we developed a disposable easy-to-use electrochemical microfluidic chip combined with multiple antibodies against six kinds of biomarkers (carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), Helicobacter pylori CagA protein (H.P.), P53oncoprotein (P53), pepsinogen I (PG I), and PG-II). The six kinds of biomarkers related to gastric cancer can be detected sensitively and synchronously in a short time. The specially designed three electrodes system enables cross-contamination to be avoided effectively. The linear ranges of detection of the electrochemical microfluidic chip were as follows: 0.37-90 ng mL(-1) for CEA, 10.75-172 U mL(-1) for CA19-9, 10-160 U L(-1) for H.P., 35-560 ng mL(-1) for P53, 37.5-600 ng mL(-1) for PG I, and 2.5-80 ng mL(-1)for PG II. This method owns better sensitivity compared with enzyme-linked immunosorbent assay (ELISA) results of 394 specimens of gastric cancer sera. Furthermore, we established a multi-index prediction model based on the six kinds of biomarkers for predicting risk of gastric cancer. In conclusion, the electrochemical microfluidic chip for detecting multiple biomarkers has great potential in applications such as early screening of gastric cancer patients, and therapeutic evaluation, and real-time dynamic monitoring the progress of gastric cancer in near future.

  5. Investigating Nonalcoholic Fatty Liver Disease in a Liver-on-a-Chip Microfluidic Device

    PubMed Central

    Simonelli, Maria Chiara; Giannitelli, Sara Maria; Businaro, Luca; Trombetta, Marcella; Rainer, Alberto

    2016-01-01

    Background and Aim Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease worldwide, ranging from simple steatosis to nonalcoholic steatohepatitis, which may progress to cirrhosis, eventually leading to hepatocellular carcinoma (HCC). HCC ranks as the third highest cause of cancer-related death globally, requiring an early diagnosis of NAFLD as a potential risk factor. However, the molecular mechanisms underlying NAFLD are still under investigation. So far, many in vitro studies on NAFLD have been hampered by the limitations of 2D culture systems, in which cells rapidly lose tissue-specific functions. The present liver-on-a-chip approach aims at filling the gap between conventional in vitro models, often scarcely predictive of in vivo conditions, and animal models, potentially biased by their xenogeneic nature. Methods HepG2 cells were cultured into a microfluidically perfused device under free fatty acid (FFA) supplementation, namely palmitic and oleic acid, for 24h and 48h. The device mimicked the endothelial-parenchymal interface of a liver sinusoid, allowing the diffusion of nutrients and removal of waste products similar to the hepatic microvasculature. Assessment of intracellular lipid accumulation, cell viability/cytotoxicity and oxidative stress due to the FFA overload, was performed by high-content analysis methodologies using fluorescence-based functional probes. Results The chip enables gradual and lower intracellular lipid accumulation, higher hepatic cell viability and minimal oxidative stress in microfluidic dynamic vs. 2D static cultures, thus mimicking the chronic condition of steatosis observed in vivo more closely. Conclusions Overall, the liver-on-a-chip system provides a suitable culture microenvironment, representing a more reliable model compared to 2D cultures for investigating NAFLD pathogenesis. Hence, our system is amongst the first in vitro models of human NAFLD developed within a microfluidic device in a sinusoid

  6. Alternating Current Cloud Point Extraction on a Microfluidic Chip: the Use of Ferrocenyl Surfactants.

    PubMed

    Usui, Yuya; Sasaki, Naoki

    2016-01-01

    Alternating current cloud point extraction (ACPE) is a preconcentration technique that can be employed in the analysis of membrane proteins on a microfluidic chip. However, the selectivity of ACPE relies on the hydrophobicity of the analytes. In this study, 11-ferrocenyltrimethylundecylammonium bromide (FTMA) was utilized to introduce electrostatic interaction as part of the ACPE technique. The use of ACPE with oxidized FTMA resulted in efficient concentration of fluorescently labeled anionic membrane proteins. We expect the approach outlined in this report to be useful in the preconcentration technique of microchip electrophoresis.

  7. Effect of the gelation process on the production of alginate microbeads by microfluidic chip technology.

    PubMed

    Capretto, Lorenzo; Mazzitelli, Stefania; Balestra, Cosimo; Tosi, Azzura; Nastruzzi, Claudio

    2008-04-01

    The present paper reports the production of Ba-alginate microspheres by microfluidic chip technology. The general production strategy is based on the formation of an alginate multiphase flow by a 'Y' junction squeezing mechanism. Special emphasis is given to the relationship existing between the gelation process and the final morphological characteristics of the produced microbeads. A series of different gelation strategies, namely: 'external gelation', 'internal gelation' and 'partial gelation' were compared in terms of size, size distribution and morphology of the produced microbeads.

  8. Gold nanorod-facilitated localized heating of droplets in microfluidic chips.

    PubMed

    Li, Zhiyong; Wang, Pan; Tong, Limin; Zhang, Lei

    2013-01-14

    A gold nanorod-facilitated optical heating method for droplets in microfluidic chips is reported. Individual and stream nanoliter level droplets containing gold nanorods are heated by a low power 808-nm-wavelength laser. Owing to the high photothermal conversion efficiency of gold nanorods, a droplet temperature of 95 °C is achieved by employing a 13.6 mW laser with good reproducibility. The heating and cooling times are 200 and 800 ms, respectively, which are attributed to the fast thermal-transfer rates of the droplets. By controlling the irradiation laser power, the temperature cycles for polymerase chain reaction are also demonstrated.

  9. A Simple Microfluidic Chip Design for Fundamental Bioseparation

    PubMed Central

    Chan, Alan S.; Danquah, Michael K.; Hartley, Patrick G.; Zhu, Yonggang

    2014-01-01

    A microchip pressure-driven liquid chromatographic system with a packed column has been designed and fabricated by using poly(dimethylsiloxane) (PDMS). The liquid chromatographic column was packed with mesoporous silica beads of Ia3d space group. Separation of dyes and biopolymers was carried out to verify the performance of the chip. A mixture of dyes (fluorescein and rhodamine B) and a biopolymer mixture (10 kDa Dextran and 66 kDa BSA) were separated and the fluorescence technique was employed to detect the movement of the molecules. Fluorescein molecule was a nonretained species and rhodamine B was attached onto silica surface when dye mixture in deionized water was injected into the microchannel. The retention times for dextran molecule and BSA molecule in biopolymer separation experiment were 45 s and 120 s, respectively. Retention factor was estimated to be 3.3 for dextran and 10.4 for BSA. The selectivity was 3.2 and resolution was 10.7. Good separation of dyes and biopolymers was achieved and the chip design was verified. PMID:24527255

  10. On-chip gradient generation in 256 microfluidic cell cultures: simulation and experimental validation.

    PubMed

    Somaweera, Himali; Haputhanthri, Shehan O; Ibraguimov, Akif; Pappas, Dimitri

    2015-08-07

    A microfluidic diffusion diluter was used to create a stable concentration gradient for dose response studies. The microfluidic diffusion diluter used in this study consisted of 128 culture chambers on each side of the main fluidic channel. A calibration method was used to find unknown concentrations with 12% error. Flow rate dependent studies showed that changing the flow rates generated different gradient patterns. Mathematical simulations using COMSOL Multi-physics were performed to validate the experimental data. The experimental data obtained for the flow rate studies agreed with the simulation results. Cells could be loaded into culture chambers using vacuum actuation and cultured for long times under low shear stress. Decreasing the size of the culture chambers resulted in faster gradient formation (20 min). Mass transport into the side channels of the microfluidic diffusion diluter used in this study is an important factor in creating the gradient using diffusional mixing as a function of the distance. To demonstrate the device's utility, an H2O2 gradient was generated while culturing Ramos cells. Cell viability was assayed in the 256 culture chambers, each at a discrete H2O2 concentration. As expected, the cell viability for the high concentration side channels increased (by injecting H2O2) whereas the cell viability in the low concentration side channels decreased along the chip due to diffusional mixing as a function of distance. COMSOL simulations were used to identify the effective concentration of H2O2 for cell viability in each side chamber at 45 min. The gradient effects were confirmed using traditional H2O2 culture experiments. Viability of cells in the microfluidic device under gradient conditions showed a linear relationship with the viability of the traditional culture experiment. Development of the microfluidic device used in this study could be used to study hundreds of concentrations of a compound in a single experiment.

  11. Microfluidic Devices for Terahertz Spectroscopy of Live Cells Toward Lab-on-a-Chip Applications.

    PubMed

    Tang, Qi; Liang, Min; Lu, Yi; Wong, Pak Kin; Wilmink, Gerald J; Zhang, Donna; Xin, Hao

    2016-04-04

    THz spectroscopy is an emerging technique for studying the dynamics and interactions of cells and biomolecules, but many practical challenges still remain in experimental studies. We present a prototype of simple and inexpensive cell-trapping microfluidic chip for THz spectroscopic study of live cells. Cells are transported, trapped and concentrated into the THz exposure region by applying an AC bias signal while the chip maintains a steady temperature at 37 °C by resistive heating. We conduct some preliminary experiments on E. coli and T-cell solution and compare the transmission spectra of empty channels, channels filled with aqueous media only, and channels filled with aqueous media with un-concentrated and concentrated cells.

  12. Microfluidic Devices for Terahertz Spectroscopy of Live Cells Toward Lab-on-a-Chip Applications

    PubMed Central

    Tang, Qi; Liang, Min; Lu, Yi; Wong, Pak Kin; Wilmink, Gerald J.; D. Zhang, Donna; Xin, Hao

    2016-01-01

    THz spectroscopy is an emerging technique for studying the dynamics and interactions of cells and biomolecules, but many practical challenges still remain in experimental studies. We present a prototype of simple and inexpensive cell-trapping microfluidic chip for THz spectroscopic study of live cells. Cells are transported, trapped and concentrated into the THz exposure region by applying an AC bias signal while the chip maintains a steady temperature at 37 °C by resistive heating. We conduct some preliminary experiments on E. coli and T-cell solution and compare the transmission spectra of empty channels, channels filled with aqueous media only, and channels filled with aqueous media with un-concentrated and concentrated cells. PMID:27049392

  13. Measurement of Single-Cell Deformability Using Impedance Analysis on Microfluidic Chip

    NASA Astrophysics Data System (ADS)

    Kim, Dongil; Choi, Eunpyo; Choi, Sung Sik; Lee, Sangho; Park, Jungyul; Yun, Kwang-Seok

    2010-12-01

    In this paper, we propose a microfluidic chip that measures the deformability of single cells by an impedance measurement method. The proposed chip is designed to differentiate the deformability of various cells by measuring the length of their stretched membrane indirectly according to the variation of the impedance after applying aspiration pressure to the cell membrane. The length of the stretched cell membrane is proportional to the applied pressure. Lengths of 18 and 21 µm were observed at the same suction pressure for human breast normal cells (MCF-10A) and caner cells (MCF-7), respectively. Electrical measurement was performed using an impedance analyzer at various frequencies. Results revealed that the impedance measurement method can be used to analyze the biomechanical characteristics of single cells, which indicates the state of malignancy of cells.

  14. Fabrication of microfluidic chips using lithographic patterning and adhesive bonding of the thick negative photoresist AZ 125 nXT

    NASA Astrophysics Data System (ADS)

    Knoll, Thorsten; Bergmann, Andreas; Nußbaum, Dominic

    2015-05-01

    In this work, for the first time the negative photoresist AZ 125 nXT was used for the fabrication of a microfluidic chip. Usually, fabrication of microfluidic devices on the basis of silicon or glass substrates is done by using the epoxy-based negative photoresist SU-8 or other thick film polymer materials. The suitability of SU-8 for various microfluidic applications has been shown in the fields of bioanalytic devices, lab-on-chip systems or microreaction technology. However, processing is always a very challenging task with regard to the adaptation of process parameters to the individual design and required functionality. Now, the AZ 125 nXT allows for the fabrication of structures in a wide thickness range with only one type of viscosity. In contrast to SU-8, the AZ 125 nXT is fully cross-linked during UV exposure and does not require a time-consuming post-exposure bake. 90 μm deep microfluidic channels were defined by lithographic patterning of AZ 125 nXT. Sealing of the open microfluidic channels was performed by a manual adhesive bonding process at a temperature of 100 °C. The fluidic function was successfully tested with flow rates up to 20 ml/min by means of a microfluidic edge connector. Long term stability and chemical resistance of the fabricated microfluidic channels will be investigated in the near future. The presented work shows the potential of AZ 125 nXT as a possible alternative to SU-8 for the fabrication of microfluidic chips.

  15. Miniature interferometer for refractive index measurement in microfluidic chip

    NASA Astrophysics Data System (ADS)

    Chen, Minghui; Geiser, Martial; Truffer, Frederic; Song, Chengli

    2012-12-01

    The design and development of the miniaturized interferometer for measurement of the refractive index or concentration of sub-microliter volume aqueous solution in microfludic chip is presented. It is manifested by a successful measurement of the refractive index of sugar-water solution, by utilizing a laser diode for light source and the small robust instrumentation for practical implementation. Theoretically, the measurement principle and the feasibility of the system are analyzed. Experimental device is constructed with a diode laser, lens, two optical plate and a complementary metal oxide semiconductor (CMOS). Through measuring the positional changes of the interference fringes, the refractive index change are retrieved. A refractive index change of 10-4 is inferred from the measured image data. The entire system is approximately the size of half and a deck of cards and can operate on battery power for long time.

  16. Novel microfluidic platform for automated lab-on-chip testing of hypercoagulability panel.

    PubMed

    Emani, Sirisha; Sista, Ramakrishna; Loyola, Hugo; Trenor, Cameron C; Pamula, Vamsee K; Emani, Sitaram M

    2012-12-01

    Current methods for hypercoagulability panel testing require large blood volumes and long turn-around testing times. A novel microfluidic platform has been designed to perform automated multiplexed hypercoagulability panel testing at near patient, utilizing only a single droplet of blood sample. We test the hypothesis that this novel platform could be utilized to perform specific multiplexed ELISA-based hypercoagulability panel testing for antithrombin III, protein C, protein S and factor VIII antigens, as well as anticardiolipin/human anti-β2-glycoprotein-1 IgG antibodies--on blood samples. Sandwich ELISA was modified by utilizing magnetic beads coated with specific antibodies as the solid phase using fluorescence readout. Percentage recovery was calculated using four-parameter logistic curves. On-chip ELISA with single factors was compared with multiplex factor ELISA for known concentrations of sample. Blood samples were analyzed on-chip and compared with traditional bench-top assays. Time for multiplexed performance of hypercoagulability panel ELISA on-chip with controls is 72 min. Recovery rates (range 80-120%) for known concentrations of specific factors was not significantly different when assays were performed using a single factor vs. multiplex factor analysis. Assay results were not significantly different between individual assays performed either on bench-top or on-chip with patient blood and/or plasma. Utilizing a novel digital microfluidic platform, we demonstrate the feasibility of automated hypercoagulability panel testing on small volume of plasma and whole blood patient samples with high fidelity. Further investigation is required to test the application of this novel technology at point-of-care clinical settings.

  17. All inkjet-printed electroactive polymer actuators for microfluidic lab-on-chip systems

    NASA Astrophysics Data System (ADS)

    Pabst, Oliver; Beckert, Erik; Perelaer, Jolke; Schubert, Ulrich S.; Eberhardt, Ramona; Tünnermann, Andreas

    2013-04-01

    Piezoelectric electroactive polymers (EAP) are promising materials for applications in microfluidic lab-on-chip systems. In such systems, fluids can be analyzed by different chemical or physical methods. During the analysis the fluids need to be distributed through the channels of the chip, which requires a pumping function. We present here all inkjet-printed EAP actuators that can be configured as a membrane-based micropump suitable for direct integration into lab-on-chip systems. Drop-on-demand inkjet printing is a versatile digital deposition technique that is capable of depositing various functional materials onto a wide variety of substrates in an additive way. Compared to conventional lithography-based processing it is cost-efficient and flexible, as no masking is required. The actuators consist of a polymer foil substrate with an inkjet-printed EAP layer sandwiched between a set of two electrodes. The actuators are printed using a commercially available EAP solution and silver nanoparticle inks. When a voltage is applied across the polymer layer, piezoelectric strain leads to a bending deflection of the beam or membrane. Circular membrane actuators with 20 mm diameter and EAP thicknesses of 10 to 15 μm exhibit deflections of several μm when driven at their resonance frequency with voltages of 110 V. From the behavior of membrane actuators a pumping rate of several 100 μL/min can be estimated, which is promising for applications in lab-on-chip devices.

  18. Fabrication of a polystyrene microfluidic chip coupled to electrospray ionization mass spectrometry for protein analysis.

    PubMed

    Hu, Xianqiao; Dong, Yuanyuan; He, Qiaohong; Chen, Hengwu; Zhu, Zhiwei

    2015-05-15

    A highly integrated polystyrene (PS) microfluidic chip coupled to electrospray ionization mass spectrometry for on-chip protein digestion and online analysis was developed. The immobilized enzymatic microreactor for on-chip protein digestion was integrated onto microchip via the novel method of region-selective UV-modification combined with glutaraldehyde-based immobilization. The micro film electric contact for applying high voltage was prepared on chips by using UV-directed electroless plating technique. A micro-tip was machined at the end of main channel, serving as the interface between microchip and mass spectrometric detector. On-chip digestion and online detection of protein was carried out by coupling the microchip with mass spectrometry (MS). The influences of methanol flow rate in side channel on the stability of spray and intensity of signals were investigated systematically. Also the influence of sample flow rate on the performance of immobilized enzymatic reactor were investigated. Stable spray was obtained at the spray voltage of 2.8-3.0kV and the methanol flow rate of 500-700nLmin(-1) with the relative standard deviation (RSD) of total ion current (TIC) less than 10%. The influence of sample flow rate on the performance of immobilized enzymatic reactor was also studied. The sequence coverage of protein identification decreased with the increase of flow rate of the sample solution. A sequence coverage of 96% was obtained with immobilized enzymatic reactor at the sample flow rate of 100nLmin(-1) with the reaction time of 8.4min. It could detect cytochrome c as low as 10μgmL(-1) with the developed system. No obvious decrease in protein digestion efficiency was observed after the chip continuously performed for 4h and stored for 15d.

  19. Microfluidic Arrayed Lab-On-A-Chip for Electrochemical Capacitive Detection of DNA Hybridization Events.

    PubMed

    Ben-Yoav, Hadar; Dykstra, Peter H; Bentley, William E; Ghodssi, Reza

    2017-01-01

    A microfluidic electrochemical lab-on-a-chip (LOC) device for DNA hybridization detection has been developed. The device comprises a 3 × 3 array of microelectrodes integrated with a dual layer microfluidic valved manipulation system that provides controlled and automated capabilities for high throughput analysis of microliter volume samples. The surface of the microelectrodes is functionalized with single-stranded DNA (ssDNA) probes which enable specific detection of complementary ssDNA targets. These targets are detected by a capacitive technique which measures dielectric variation at the microelectrode-electrolyte interface due to DNA hybridization events. A quantitative analysis of the hybridization events is carried out based on a sensing modeling that includes detailed analysis of energy storage and dissipation components. By calculating these components during hybridization events the device is able to demonstrate specific and dose response sensing characteristics. The developed microfluidic LOC for DNA hybridization detection offers a technology for real-time and label-free assessment of genetic markers outside of laboratory settings, such as at the point-of-care or in-field environmental monitoring.

  20. Continuous Flow 1H and 13C NMR Spectroscopy in Microfluidic Stripline NMR Chips

    PubMed Central

    2017-01-01

    Microfluidic stripline NMR technology not only allows for NMR experiments to be performed on small sample volumes in the submicroliter range, but also experiments can easily be performed in continuous flow because of the stripline’s favorable geometry. In this study we demonstrate the possibility of dual-channel operation of a microfluidic stripline NMR setup showing one- and two-dimensional 1H, 13C and heteronuclear NMR experiments under continuous flow. We performed experiments on ethyl crotonate and menthol, using three different types of NMR chips aiming for straightforward microfluidic connectivity. The detection volumes are approximately 150 and 250 nL, while flow rates ranging from 0.5 μL/min to 15 μL/min have been employed. We show that in continuous flow the pulse delay is determined by the replenishment time of the detector volume, if the sample trajectory in the magnet toward NMR detector is long enough to polarize the spin systems. This can considerably speed up quantitative measurement of samples needing signal averaging. So it can be beneficial to perform continuous flow measurements in this setup for analysis of, e.g., reactive, unstable, or mass-limited compounds. PMID:28194934

  1. Continuous Flow (1)H and (13)C NMR Spectroscopy in Microfluidic Stripline NMR Chips.

    PubMed

    Oosthoek-de Vries, Anna Jo; Bart, Jacob; Tiggelaar, Roald M; Janssen, Johannes W G; van Bentum, P Jan M; Gardeniers, Han J G E; Kentgens, Arno P M

    2017-02-21

    Microfluidic stripline NMR technology not only allows for NMR experiments to be performed on small sample volumes in the submicroliter range, but also experiments can easily be performed in continuous flow because of the stripline's favorable geometry. In this study we demonstrate the possibility of dual-channel operation of a microfluidic stripline NMR setup showing one- and two-dimensional (1)H, (13)C and heteronuclear NMR experiments under continuous flow. We performed experiments on ethyl crotonate and menthol, using three different types of NMR chips aiming for straightforward microfluidic connectivity. The detection volumes are approximately 150 and 250 nL, while flow rates ranging from 0.5 μL/min to 15 μL/min have been employed. We show that in continuous flow the pulse delay is determined by the replenishment time of the detector volume, if the sample trajectory in the magnet toward NMR detector is long enough to polarize the spin systems. This can considerably speed up quantitative measurement of samples needing signal averaging. So it can be beneficial to perform continuous flow measurements in this setup for analysis of, e.g., reactive, unstable, or mass-limited compounds.

  2. Soil-on-a-Chip: microfluidic platforms for environmental organismal studies.

    PubMed

    Stanley, Claire E; Grossmann, Guido; i Solvas, Xavier Casadevall; deMello, Andrew J

    2016-01-21

    Soil is the habitat of countless organisms and encompasses an enormous variety of dynamic environmental conditions. While it is evident that a thorough understanding of how organisms interact with the soil environment may have substantial ecological and economical impact, current laboratory-based methods depend on reductionist approaches that are incapable of simulating natural diversity. The application of Lab-on-a-Chip or microfluidic technologies to organismal studies is an emerging field, where the unique benefits afforded by system miniaturisation offer new opportunities for the experimentalist. Indeed, precise spatiotemporal control over the microenvironments of soil organisms in combination with high-resolution imaging has the potential to provide an unprecedented view of biological events at the single-organism or single-cell level, which in turn opens up new avenues for environmental and organismal studies. Herein we review some of the most recent and interesting developments in microfluidic technologies for the study of soil organisms and their interactions with the environment. We discuss how so-called "Soil-on-a-Chip" technology has already contributed significantly to the study of bacteria, nematodes, fungi and plants, as well as inter-organismal interactions, by advancing experimental access and environmental control. Most crucially, we highlight where distinct advantages over traditional approaches exist and where novel biological insights will ensue.

  3. A Butyl Methacrylate Monolithic Column Prepared In-Situ on a Microfluidic Chip and its Applications

    PubMed Central

    Xu, Yi; Zhang, Wenpin; Zeng, Ping; Cao, Qiang

    2009-01-01

    A butyl methacrylate (BMA) monolithic column was polymerized in-situ with UV irradiation in an ultraviolet transparent PDMS micro-channel on a homemade micro-fluidic chip. Under the optimized conditions and using a typical polymerization mixture consisting of 75% porogenic solvents and 25% monomers, the BMA monolithic column was obtained as expected. The BET surface area ratio of the BMA monolithic column was 366 m2·g-1. The corresponding SEM images showed that the monolithic column material polymerized in a glass channel was composed of uniform pores and spherical particles with diameters ranging from 3 to 5 μm. The promethazine–luminal–potassium ferricyanide chemiluminescence system was selected for testing the capability of the column. A flow injection analytical technique–chemiluminescence (FIA–CL) system on the microfluidic chip with a BMA monolithic column pretreatment unit was established to determine promethazine. Trace promethazine was enriched by the BMA monolithic column, with more than a 10-fold average enrichment ratio. The proposed method has a linear response concentration range of 1.0×10-8 - 1.0×10-6g·mL-1 and the detection limit was 1.6×10-9g·mL-1. PMID:22412320

  4. Recent advancements in chemical luminescence-based lab-on-chip and microfluidic platforms for bioanalysis.

    PubMed

    Mirasoli, Mara; Guardigli, Massimo; Michelini, Elisa; Roda, Aldo

    2014-01-01

    Miniaturization of analytical procedures through microchips, lab-on-a-chip or micro total analysis systems is one of the most recent trends in chemical and biological analysis. These systems are designed to perform all the steps in an analytical procedure, with the advantages of low sample and reagent consumption, fast analysis, reduced costs, possibility of extra-laboratory application. A range of detection technologies have been employed in miniaturized analytical systems, but most applications relied on fluorescence and electrochemical detection. Chemical luminescence (which includes chemiluminescence, bioluminescence, and electrogenerated chemiluminescence) represents an alternative detection principle that offered comparable (or better) analytical performance and easier implementation in miniaturized analytical devices. Nevertheless, chemical luminescence-based ones represents only a small fraction of the microfluidic devices reported in the literature, and until now no review has been focused on these devices. Here we review the most relevant applications (since 2009) of miniaturized analytical devices based on chemical luminescence detection. After a brief overview of the main chemical luminescence systems and of the recent technological advancements regarding their implementation in miniaturized analytical devices, analytical applications are reviewed according to the nature of the device (microfluidic chips, microchip electrophoresis, lateral flow- and paper-based devices) and the type of application (micro-flow injection assays, enzyme assays, immunoassays, gene probe hybridization assays, cell assays, whole-cell biosensors).

  5. Microfluidic integration of wirebonded microcoils for on-chip applications in nuclear magnetic resonance

    NASA Astrophysics Data System (ADS)

    Meier, Robert Ch; Höfflin, Jens; Badilita, Vlad; Wallrabe, Ulrike; Korvink, Jan G.

    2014-04-01

    We present an integrated microfluidic device for on-chip nuclear magnetic resonance (NMR) studies of microscopic samples. The devices are fabricated by means of a MEMS compatible process, which joins the automatic wirebond winding of solenoidal microcoils and the manufacturing of a complex microfluidic network using dry-photoresist lamination. The wafer-scale cleanroom process is potentially capable of mass fabrication. Since the non-invasive NMR analysis technique is rather insensitive, particularly when microscopic sample volumes are to be investigated, we also focus on the optimization of the wirebonded microcoil for this purpose. The on-chip measurement of NMR signals from a 20 nl sample are evaluated for imaging analysis of microparticles, as well as for spectroscopy. Whereas the latter revealed that the sensitivity of the MEMS microcoil is comparable with hand-wound devices and achieves a full-width-half-maximum linewidth of 8 Hz, the imaging experiment demonstrated 10 μm isotropic spatial resolution within an experiment time of 38 min for a 3D image with a field of view of 1 mm × 1 mm × 0.5 mm (500 000 voxels).

  6. Ultrasmall volume molecular isothermal amplification in microfluidic chip with advanced surface processing

    NASA Astrophysics Data System (ADS)

    Huang, Guoliang; Ma, Li; Yang, Xiaoyong; Yang, Xu

    2011-01-01

    In this paper, we developed a metal micro-fluidic chip with advanced surface processing for ultra-small volume molecular isothermal amplification. This method takes advantages of the nucleic acid amplification with good stability and consistency, high sensitivity about 31 genomic DNA copies and bacteria specific gene identification. Based on the advanced surface processing, the bioreaction assays of nucleic acid amplification was dropped about 392nl in volume. A high numerical aperture confocal optical detection system was advanced to sensitively monitor the DNA amplification with low noise and high power collecting fluorescence near to the optical diffraction limit. A speedy nucleic acid isothermal amplification was performed in the ultra-small volume microfluidic chip, where the time at the inflexions of second derivative to DNA exponential amplified curves was brought forward and the sensitivity was improved about 65 folds to that of in current 25μl Ep-tube amplified reaction, which indicates a promising clinic molecular diagnostics in the droplet amplification.

  7. A microfluidic device for on-chip agarose microbead generation with ultralow reagent consumption.

    PubMed

    Desbois, Linda; Padirac, Adrien; Kaneda, Shohei; Genot, Anthony J; Rondelez, Yannick; Hober, Didier; Collard, Dominique; Fujii, Teruo

    2012-01-01

    Water-in-oil microdroplets offer microreactors for compartmentalized biochemical reactions with high throughput. Recently, the combination with a sol-gel switch ability, using agarose-in-oil microdroplets, has increased the range of possible applications, allowing for example the capture of amplicons in the gel phase for the preservation of monoclonality during a PCR reaction. Here, we report a new method for generating such agarose-in-oil microdroplets on a microfluidic device, with minimized inlet dead volume, on-chip cooling, and in situ monitoring of biochemical reactions within the gelified microbeads. We used a flow-focusing microchannel network and successfully generated agarose microdroplets at room temperature using the "push-pull" method. This method consists in pushing the oil continuous phase only, while suction is applied to the device outlet. The agarose phase present at the inlet is thus aspirated in the device, and segmented in microdroplets. The cooling system consists of two copper wires embedded in the microfluidic device. The transition from agarose microdroplets to microbeads provides additional stability and facilitated manipulation. We demonstrate the potential of this method by performing on-chip a temperature-triggered DNA isothermal amplification in agarose microbeads. Our device thus provides a new way to generate microbeads with high throughput and no dead volume for biochemical applications.

  8. Surface Roughness Study on Microchannels of CO2 Laser Fabricating Pmma-Based Microfluidic Chip

    NASA Astrophysics Data System (ADS)

    Chen, Xueye; Li, Tiechuan; Fu, Baoding

    A novel method named soak sacrificial layer ultrasonic method (SSLUM) has been presented for optimizing the surface roughness of the microchannels of polymethyl methacrylate (PMMA)-based microfluidic chips. CO2 laser was used for ablative microchannels on the PMMA sheet, and the effects of key parameters including laser power, laser ablation speed and solution concentration on the surface roughness of microchannels were estimated and optimized by SSLUM. The experimental observation demonstrates that the surface roughness results mainly from the residues on the channel wall, which are produced by the bubbles movement and bursting. The research results show that the surface roughness can be improved effectively by using SSLUM. In our experiment, the best value was Ra = 110nm with laser power 12W, laser ablation speed 10mm/s, the solution concentration 75%, and the time of ultrasonic vibration 25min. SSLUM is proven to be an effective, simple and rapid method for optimizing the surface roughness of microchannels of microfluidic chips.

  9. Detection of DNAs by Using Dual Packed Polystyrene Bead-Quantum Dots in a Microfluidic Chip.

    PubMed

    Le, Ngoc Tam; Kim, Jong Sung

    2015-01-01

    The semiconductor nanocrystals (or quantum dots) have shown peculiar optical and electrical properties due to their exceptionally small size. In recent years, tremendous researches on quantum dots have been carried out. Among them, QDs as sensing media for biological assay have achieved a great progress. Recently we have reported the detection of DNAs by using fluorescence quenching of QDs after DNA hybridization. Several oligonucleotides and human genomic genes could be detected. In this report we used dual packing of polystyrene bead-quantum dots to detect different kinds of DNAs simultaneously. QDs with different emission peaks were used. Carboxylated-CdSe/ZnS QDs (emission: 525, 605 nm) could bind to microbeads of polystyrene/divinyl benzene via EDC/NHS cross-linking reaction. Polystyrene bead-QDs with different colors were packed in the channel of the microfluidic chip. The fluorescence quenching from the QDs by intercalating dye was observed after hybridization of exon 6 and 7 of p53 gene at the weir in the channel of microfluidic chip. The simultaneous fluorescence quenching of the QDs by PI and TOTO-3 were observed.

  10. The Evopopbot Chip: Ultra High-throughput Evolutionary Population Bottlenecking using Drop-Based Microfluidics

    NASA Astrophysics Data System (ADS)

    Chang, Connie; Rotem, Assaf; Serohijos, Adrian; Zhang, Huidan; Tao, Ye; Fischer Hesselbrock, Audrey; Thielen, Peter; Mehoke, Thomas; Wolfe, Joshua; Wobus, Christiane; Feldman, Andrew; Shakhnovich, Eugene; Weitz, David

    2014-03-01

    The study of how viruses propagate is important for curing disease and preventing viral outbreaks. In nature, viruses can compete with one another, and the most evolutionary fit virus usually takes over a population. Yet there exist variants in the population that can escape subjected evolutionary pressures and eventually dominate the population. Successful studies of viral epidemics hinges on the ability to access these variants. Here, we present the use of droplet-based microfluidics as a simple method to segregate and propagate a viral population as individual viral lineages, simultaneously performing millions of in vitroevolutionary bottlenecking experiments. We introduce a novel microfluidic device, called the ``Evopopbot Chip'', that allows for simultaneous passaging of millions of evolutionary bottlenecking events by splitting drops containing previous generations of viruses and merging with drops containing new host cells. After several generations of viral replication in the evolution chip, we discover hundreds of new viruses that are able to escape a neutralizing antibody selection pressure compared to bulk passaging.

  11. Hot embossed polyethylene through-hole chips for bead-based microfluidic devices

    PubMed Central

    Chou, Jie; Du, Nan; Ou, Tina; Floriano, Pierre N.; Christodoulides, Nicolaos; McDevitt, John T.

    2013-01-01

    Over the past decade, there has been a growth of interest in the translation of microfluidic systems into real-world clinical practice, especially for use in point-of-care or near patient settings. While initial fabrication advances in microfluidics involved mainly the etching of silicon and glass, the economics of scaling of these materials is not amendable for point-of-care usage where single-test applications forces cost considerations to be kept low and throughput high. As such, a materials base more consistent with point-of-care needs is required. In this manuscript, the fabrication of a hot embossed, through-hole low-density polyethylene ensembles derived from an anisotropically etched silicon wafer is discussed. This semi-opaque polymer that can be easily sterilized and recycled provides low background noise for fluorescence measurements and yields more affordable cost than other thermoplastics commonly used for microfluidic applications such as cyclic olefin copolymer (COC). To fabrication through-hole microchips from this alternative material for microfluidics, a fabrication technique that uses a high-temperature, high-pressure resistant mold is described. This aluminum-based epoxy mold, serving as the positive master mold for embossing, is casted over etched arrays of pyramidal pits in a silicon wafer. Methods of surface treatment of the wafer prior to casting and PDMS casting of the epoxy are discussed to preserve the silicon wafer for future use. Changes in the thickness of polyethylene are observed for varying embossing temperatures. The methodology described herein can quickly fabricate 20 disposable, single use chips in less than 30 minutes with the ability to scale up 4x by using multiple molds simultaneously. When coupled as a platform supporting porous bead sensors, as in the recently developed Programmable Bio-Nano-Chip, this bead chip system can achieve limits of detection, for the cardiac biomarker C-reactive protein, of 0.3 ng/mL, thereby

  12. Hot embossed polyethylene through-hole chips for bead-based microfluidic devices.

    PubMed

    Chou, Jie; Du, Nan; Ou, Tina; Floriano, Pierre N; Christodoulides, Nicolaos; McDevitt, John T

    2013-04-15

    Over the past decade, there has been a growth of interest in the translation of microfluidic systems into real-world clinical practice, especially for use in point-of-care or near patient settings. While initial fabrication advances in microfluidics involved mainly the etching of silicon and glass, the economics of scaling of these materials is not amendable for point-of-care usage where single-test applications force cost considerations to be kept low and throughput high. As such, materials base more consistent with point-of-care needs is required. In this manuscript, the fabrication of a hot embossed, through-hole low-density polyethylene ensembles derived from an anisotropically etched silicon wafer is discussed. This semi-opaque polymer that can be easily sterilized and recycled provides low background noise for fluorescence measurements and yields more affordable cost than other thermoplastics commonly used for microfluidic applications such as cyclic olefin copolymer (COC). To fabrication through-hole microchips from this alternative material for microfluidics, a fabrication technique that uses a high-temperature, high-pressure resistant mold is described. This aluminum-based epoxy mold, serving as the positive master mold for embossing, is casted over etched arrays of pyramidal pits in a silicon wafer. Methods of surface treatment of the wafer prior to casting and PDMS casting of the epoxy are discussed to preserve the silicon wafer for future use. Changes in the thickness of polyethylene are observed for varying embossing temperatures. The methodology described herein can quickly fabricate 20 disposable, single use chips in less than 30 min with the ability to scale up 4 times by using multiple molds simultaneously. When coupled as a platform supporting porous bead sensors, as in the recently developed Programmable Bio-Nano-Chip, this bead chip system can achieve limits of detection, for the cardiac biomarker C-reactive protein, of 0.3 ng/mL, thereby

  13. Detection of influenza virus using a lateral flow immunoassay for amplified DNA by a microfluidic RT-PCR chip.

    PubMed

    Nagatani, Naoki; Yamanaka, Keiichiro; Ushijima, Hiromi; Koketsu, Ritsuko; Sasaki, Tadahiro; Ikuta, Kazuyoshi; Saito, Masato; Miyahara, Toshiro; Tamiya, Eiichi

    2012-08-07

    Influenza virus RNA was amplified by a continuous-flow polydimethylsiloxane microfluidic RT-PCR chip within 15-20 min. The amplified influenza virus RNA was observed with the naked eye, as the red color at the test line, using a lateral flow immunoassay within 1 min.

  14. An integrated sample-in-answer-out microfluidic chip for rapid human identification by STR analysis.

    PubMed

    Le Roux, Delphine; Root, Brian E; Hickey, Jeffrey A; Scott, Orion N; Tsuei, Anchi; Li, Jingyi; Saul, David J; Chassagne, Luc; Landers, James P; de Mazancourt, Philippe

    2014-11-21

    A fully integrated microfluidic chip for human identification by short tandem repeat (STR) analysis that includes a unique enzymatic liquid preparation of the DNA, microliter non-contact PCR, and a polymer that allows a high-resolution separation within a compact microchip footprint has been developed. A heat-activated enzyme that digests biological materials is employed to generate the target yield of DNA from a buccal swab or FTA paper. The microfluidic architecture meters an aliquot of the liberated DNA and mixes it with the PCR reagents prior to non-contact IR-mediated PCR amplification. The products of PCR amplification are mixed with a sizing standard (ladder) and the 18-plex STR amplicons are separated in an effective length (Leff) of just 7 cm. The development, optimization and integration of each of these processes within the microfluidic chip are described. The device is able to generate genetic profiles in approximately 2 hours that match the profiles from the conventional processes performed using separate conventional instruments. Analysis is performed on a single plastic microchip with a size similar to that of a 96-well plate and only a few mm thick with no pretreatment of any of the functional domains. This is significant advancement in terms of ease of fabrication over glass microdevices or polymeric systems assembled from multiple components. Consequently, this fully integrated sample-in-answer-out microchip is an important step toward generation of a rapid micro-total analysis system for point-of-collection human identification based on genetic analysis.

  15. Self-digitization microfluidic chip for absolute quantification of mRNA in single cells.

    PubMed

    Thompson, Alison M; Gansen, Alexander; Paguirigan, Amy L; Kreutz, Jason E; Radich, Jerald P; Chiu, Daniel T

    2014-12-16

    Quantification of mRNA in single cells provides direct insight into how intercellular heterogeneity plays a role in disease progression and outcomes. Quantitative polymerase chain reaction (qPCR), the current gold standard for evaluating gene expression, is insufficient for providing absolute measurement of single-cell mRNA transcript abundance. Challenges include difficulties in handling small sample volumes and the high variability in measurements. Microfluidic digital PCR provides far better sensitivity for minute quantities of genetic material, but the typical format of this assay does not allow for counting of the absolute number of mRNA transcripts samples taken from single cells. Furthermore, a large fraction of the sample is often lost during sample handling in microfluidic digital PCR. Here, we report the absolute quantification of single-cell mRNA transcripts by digital, one-step reverse transcription PCR in a simple microfluidic array device called the self-digitization (SD) chip. By performing the reverse transcription step in digitized volumes, we find that the assay exhibits a linear signal across a wide range of total RNA concentrations and agrees well with standard curve qPCR. The SD chip is found to digitize a high percentage (86.7%) of the sample for single-cell experiments. Moreover, quantification of transferrin receptor mRNA in single cells agrees well with single-molecule fluorescence in situ hybridization experiments. The SD platform for absolute quantification of single-cell mRNA can be optimized for other genes and may be useful as an independent control method for the validation of mRNA quantification techniques.

  16. Microfluidic Pneumatic Cages: A Novel Approach for In-chip Crystal Trapping, Manipulation and Controlled Chemical Treatment

    PubMed Central

    Abrishamkar, Afshin; Paradinas, Markos; Bailo, Elena; Rodriguez-Trujillo, Romen; Pfattner, Raphael; Rossi, René M.; Ocal, Carmen; deMello, Andrew J.; Amabilino, David B.; Puigmartí-Luis, Josep

    2016-01-01

    The precise localization and controlled chemical treatment of structures on a surface are significant challenges for common laboratory technologies. Herein, we introduce a microfluidic-based technology, employing a double-layer microfluidic device, which can trap and localize in situ and ex situ synthesized structures on microfluidic channel surfaces. Crucially, we show how such a device can be used to conduct controlled chemical reactions onto on-chip trapped structures and we demonstrate how the synthetic pathway of a crystalline molecular material and its positioning inside a microfluidic channel can be precisely modified with this technology. This approach provides new opportunities for the controlled assembly of structures on surface and for their subsequent treatment. PMID:27500740

  17. A Low-Cost Microfluidic Chip for Rapid Genotyping of Malaria-Transmitting Mosquitoes

    PubMed Central

    Liu, Changchun; Mauk, Michael G.; Hart, Robert; Bonizzoni, Mariangela; Yan, Guiyun; Bau, Haim H.

    2012-01-01

    Background Vector control is one of the most effective measures to prevent the transmission of malaria, a disease that causes over 600,000 deaths annually. Around 30–40 Anopheles mosquito species are natural vectors of malaria parasites. Some of these species cannot be morphologically distinguished, but have behavioral and ecological differences. Emblematic of this is the Anopheles gambiae species complex. The correct identification of vector species is fundamental to the development of control strategies and epidemiological studies of disease transmission. Methodology/Principal Findings An inexpensive, disposable, field-deployable, sample-to-answer, microfluidic chip was designed, constructed, and tested for rapid molecular identification of Anopheles gambiae and Anopheles arabiensis. The chip contains three isothermal amplification reactors. One test reactor operates with specific primers to amplify Anopheles gambiae DNA, another with specific primers for Anopheles arabiensis DNA, and the third serves as a negative control. A mosquito leg was crushed on an isolation membrane. Two discs, laden with mosquito tissue, were punched out of the membrane and inserted into the two test chambers. The isolated, disc-bound DNA served as a template in the amplification processes. The amplification products were detected with intercalating fluorescent dye that was excited with a blue light-emitting diode. The emitted light was observed by eye and recorded with a cell-phone camera. When the target consisted of Anopheles gambiae, the reactor containing primers specific to An. gambiae lit up while the other two reactors remained dark. When the target consisted of Anopheles arabiensis, the reactor containing primers specific to An. arabiensis lit up while the other two reactors remained dark. Conclusions/Significance The microfluidic chip provides a means to identify mosquito type through molecular analysis. It is suitable for field work, allowing one to track the geographical

  18. From bioseparation to artificial micro-organs: microfluidic chip based particle manipulation techniques

    NASA Astrophysics Data System (ADS)

    Stelzle, Martin

    2010-02-01

    Microfluidic device technology provides unique physical phenomena which are not available in the macroscopic world. These may be exploited towards a diverse array of applications in biotechnology and biomedicine ranging from bioseparation of particulate samples to the assembly of cells into structures that resemble the smallest functional unit of an organ. In this paper a general overview of chip-based particle manipulation and separation is given. In the state of the art electric, magnetic, optical and gravitational field effects are utilized. Also, mechanical obstacles often in combination with force fields and laminar flow are employed to achieve separation of particles or molecules. In addition, three applications based on dielectrophoretic forces for particle manipulation in microfluidic systems are discussed in more detail. Firstly, a virus assay is demonstrated. There, antibody-loaded microbeads are used to bind virus particles from a sample and subsequently are accumulated to form a pico-liter sized aggregate located at a predefined position in the chip thus enabling highly sensitive fluorescence detection. Secondly, subcellular fractionation of mitochondria from cell homogenate yields pure samples as was demonstrated by Western Blot and 2D PAGE analysis. Robust long-term operation with complex cell homogenate samples while avoiding electrode fouling is achieved by a set of dedicated technical means. Finally, a chip intended for the dielectrophoretic assembly of hepatocytes and endothelial cells into a structure resembling a liver sinusoid is presented. Such "artificial micro organs" are envisioned as substance screening test systems providing significantly higher predictability with respect to the in vivo response towards a substance under test.

  19. On-chip quantitative detection of pathogen genes by autonomous microfluidic PCR platform.

    PubMed

    Tachibana, Hiroaki; Saito, Masato; Shibuya, Shogo; Tsuji, Koji; Miyagawa, Nobuyuki; Yamanaka, Keiichiro; Tamiya, Eiichi

    2015-12-15

    Polymerase chain reaction (PCR)-based genetic testing has become a routine part of clinical diagnoses and food testing. In these fields, rapid, easy-to-use, and cost-efficient PCR chips are expected to be appeared for providing such testing on-site. In this study, a new autonomous disposable plastic microfluidic PCR chip was created, and was utilized for quantitative detection of pathogenic microorganisms. To control the capillary flow of the following solution in the PCR microchannel, a driving microchannel was newly designed behind the PCR microchannel. This allowed the effective PCR by simply dropping the PCR solution onto the inlet without any external pumps. In order to achieve disposability, injection-molded cyclo-olefin polymer (COP) of a cost-competitive plastic was used for the PCR chip. We discovered that coating the microchannel walls with non-ionic surfactant produced a suitable hydrophilic surface for driving the capillary flow through the 1250-mm long microchannel. As a result, quantitative real-time PCR with the lowest initial concentration of human, Escherichia coli (E. coli), and pathogenic E. coli O157 genomic DNA of 4, 0.0019, 0.031 pg/μl, respectively, was successfully achieved in less than 18 min. Our results indicate that the platform presented in this study provided a rapid, easy-to-use, and low-cost real-time PCR system that could be potentially used for on-site gene testing.

  20. Biomolecule storage on non-modified thermoplastic microfluidic chip by ink-jet printing of ionogels

    PubMed Central

    Tijero, M.; Díez-Ahedo, R.; Benito-Lopez, F.; Basabe-Desmonts, L.; Castro-López, V.; Valero, A.

    2015-01-01

    This paper reports an innovative technique for reagents storage in microfluidic devices by means of a one-step UV-photoprintable ionogel-based microarray on non-modified polymeric substrates. Although the ionogel and the ink-jet printing technology are well published, this is the first study where both are used for long-term reagent storage in lab-on-a-chip devices. This technology for reagent storage is perfectly compatible with mass production fabrication processes since pre-treatment of the device substrate is not necessary and inkjet printing allows for an efficient reagent deposition process. The functionality of this microarray is demonstrated by testing the release of biotin-647 after being stored for 1 month at room temperature. Analysis of the fluorescence of the ionogel-based microarray that contains biotin-647 demonstrated that 90% of the biotin-647 present was released from the ionogel-based microarray after pumping PBS 0.1% Tween at 37 °C. Moreover, the activity of biotin-647 after being released from the ionogel-based microarray was investigated trough the binding capability of this biotin to a microcontact printed chip surface with avidin. These findings pave the way for a novel, one-step, cheap and mass production on-chip reagents storage method applicable to other reagents such as antibodies and proteins and enzymes. PMID:26339323

  1. A microfluidic chip for formation and collection of emulsion droplets utilizing active pneumatic micro-choppers and micro-switches.

    PubMed

    Lai, Chia-Wei; Lin, Yen-Heng; Lee, Gwo-Bin

    2008-10-01

    The formation of emulsification droplets is crucial for many industrial applications. This paper reports a new microfluidic chip capable of formation and collection of micro-droplets in liquids for emulsion applications. This microfluidic chip comprising microchannels, a micro-chopper and a micro-switch was fabricated by using micro-electro-mechanical-systems (MEMS) technology. The microfluidic chip can generate uniform droplets with tunable sizes by using combination of flow-focusing and liquid-chopping techniques. The droplet size can be actively fine-tuned by controlling either the relative sheath/sample flow velocity ratios or the chopping frequency. The generated droplets can be then sorted to a specific collection area utilizing an active pneumatic micro-switch formed with three micro-valves. Experimental data showed that the olive oil and sodium-alginate (Na-alginate) droplets with diameters ranging from 3 mum to 70 mum with a variation less than 14% is successfully generated and collected. The development of this microfluidic system can be promising for emulsion, drug delivery and nano-medicine applications.

  2. Single-bead arrays for fluorescence-based immunoassays on capillary-driven microfluidic chips

    NASA Astrophysics Data System (ADS)

    Temiz, Yuksel; Lim, Michel; Delamarche, Emmanuel

    2016-03-01

    We report a concept for the simple fabrication of easy-to-use chips for immunoassays in the context of point-of-care diagnostics. The chip concept comprises mainly three features: (1) the efficient integration of reagents using beads functionalized with receptors, (2) the generation of capillary-driven liquid flows without using external pumps, and (3) a high-sensitivity detection of analytes using fluorescence microscopy. We fabricated prototype chips using dry etching of Si wafers. 4.5-μm-diameter beads were integrated into hexagonal arrays by sedimentation and removing the excess using a stream of water. We studied the effect of different parameters and showed that array occupancies from 30% to 50% can be achieved by pipetting a 250 nL droplet of 1% bead solution and allowing the beads sediment for 3 min. Chips with integrated beads were sealed using a 50-μm-thick dry-film resist laminated at 45 °C. Liquids pipetted to loading pads were autonomously pulled by capillary pumps at a rate of 0.35 nL s-1 for about 30 min. We studied ligand-receptor interactions and binding kinetics using time-lapse fluorescence microscopy and demonstrated a 5 pM limit of detection (LOD) for an anti-biotin immunoassay. As a clinically-relevant example, we implemented an immunoassay to detect prostate specific antigen (PSA) and showed an LOD of 108 fM (i.e. 3.6 pg mL-1). While a specific implementation is provided here for the detection of PSA, we believe that combining capillary-driven microfluidics with arrays of single beads and fluorescence readout to be very flexible and sufficiently sensitive for the detection of other clinically-relevant analytes.

  3. Microfluidic chips with multi-junctions: an advanced tool in recovering proteins from inclusion bodies.

    PubMed

    Yamaguchi, Hiroshi; Miyazaki, Masaya

    2015-01-01

    Active recombinant proteins are used for studying the biological functions of genes and for the development of therapeutic drugs. Overexpression of recombinant proteins in bacteria often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. Protein refolding is an important process for obtaining active recombinant proteins from inclusion bodies. However, the conventional refolding method of dialysis or dilution is time-consuming and recovered active protein yields are often low, and a cumbersome trial-and-error process is required to achieve success. To circumvent these difficulties, we used controllable diffusion through laminar flow in microchannels to regulate the denaturant concentration. This method largely aims at reducing protein aggregation during the refolding procedure. This Commentary introduces the principles of the protein refolding method using microfluidic chips and the advantage of our results as a tool for rapid and efficient recovery of active recombinant proteins from inclusion bodies.

  4. Dynamic monitoring of membrane nanotubes formation induced by vaccinia virus on a high throughput microfluidic chip

    PubMed Central

    Xiao, Min; Xu, Na; Wang, Cheng; Pang, Dai-Wen; Zhang, Zhi-Ling

    2017-01-01

    Membrane nanotubes (MNTs) are physical connections for intercellular communication and induced by various viruses. However, the formation of vaccinia virus (VACV)-induced MNTs has never been studied. In this report, VACV-induced MNTs formation process was monitored on a microfluidic chip equipped with a series of side chambers, which protected MNTs from fluidic shear stress. MNTs were formed between susceptible cells and be facilitated by VACV infection through three patterns. The formed MNTs varied with cell migration and virus concentration. The length of MNTs was positively correlated with the distance of cell migration. With increasing virus titer, the peak value of the ratio of MNT-carried cell appeared earlier. The immunofluorescence assay indicated that the rearrangement of actin fibers induced by VACV infection may lead to the formation of MNTs. This study presents evidence for the formation of MNTs induced by virus and helps us to understand the relationship between pathogens and MNTs. PMID:28317863

  5. Dynamic monitoring of membrane nanotubes formation induced by vaccinia virus on a high throughput microfluidic chip

    NASA Astrophysics Data System (ADS)

    Xiao, Min; Xu, Na; Wang, Cheng; Pang, Dai-Wen; Zhang, Zhi-Ling

    2017-03-01

    Membrane nanotubes (MNTs) are physical connections for intercellular communication and induced by various viruses. However, the formation of vaccinia virus (VACV)-induced MNTs has never been studied. In this report, VACV-induced MNTs formation process was monitored on a microfluidic chip equipped with a series of side chambers, which protected MNTs from fluidic shear stress. MNTs were formed between susceptible cells and be facilitated by VACV infection through three patterns. The formed MNTs varied with cell migration and virus concentration. The length of MNTs was positively correlated with the distance of cell migration. With increasing virus titer, the peak value of the ratio of MNT-carried cell appeared earlier. The immunofluorescence assay indicated that the rearrangement of actin fibers induced by VACV infection may lead to the formation of MNTs. This study presents evidence for the formation of MNTs induced by virus and helps us to understand the relationship between pathogens and MNTs.

  6. An integrated microfluidic chip for immunocapture, preconcentration and separation of β-amyloid peptides

    PubMed Central

    Mohamadi, Reza M.; Svobodova, Zuzana; Bilkova, Zuzana; Otto, Markus; Taverna, Myriam; Descroix, Stephanie; Viovy, Jean-Louis

    2015-01-01

    We present an integrated microfluidic chip for detection of β-amyloid (Aβ) peptides. Aβ peptides are major biomarkers for the diagnosis of Alzheimer's disease (AD) in its early stages. This microfluidic device consists of three main parts: (1) An immunocapture microcolumn based on self-assembled magnetic beads coated with antibodies specific to Aβ peptides, (2) a nano-porous membrane made of photopolymerized hydrogel for preconcentration, and (3) a microchip electrophoresis (MCE) channel with fluorescent detection. Sub-milliliter sample volume is either mixed off-chip with antibody coated magnetic beads and injected into the device or is injected into an already self-assembled column of magnetic beads in the microchannel. The captured peptides on the beads are then electrokinetically eluted and re-concentrated onto the nano-membrane in a few nano-liters. By integrating the nano-membrane, total assay time was reduced and also off-chip re-concentration or buffer exchange steps were not needed. Finally, the concentrated peptides in the chip are separated by electrophoresis in a polymer-based matrix. The device was applied to the capture and MCE analysis of differently truncated peptides Aβ (1–37, 1–39, 1–40, and 1–42) and was able to detect as low as 25 ng of synthetic Aβ peptides spiked in undiluted cerebrospinal fluid (CSF). The device was also tested with CSF samples from healthy donors. CSF samples were fluorescently labelled and pre-mixed with the magnetic beads and injected into the device. The results indicated that Aβ1-40, an important biomarker for distinguishing patients with frontotemporal lobe dementia from controls and AD patients, was detectable. Although the sensitivity of this device is not yet enough to detect all Aβ subtypes in CSF, this is the first report on an integrated or semi-integrated device for capturing and analyzing of differently truncated Aβ peptides. The method is less demanding and faster than the conventional

  7. Smart portable electrophoresis instrument based on multipurpose microfluidic chips with electrochemical detection.

    PubMed

    Fernández-la-Villa, Ana; Sánchez-Barragán, Dámaso; Pozo-Ayuso, Diego F; Castaño-Álvarez, Mario

    2012-09-01

    A second generation of a battery-powered portable electrophoresis instrument for the use of ME with electrochemical detection was developed. As the first-generation, the main unit of the instrument (150 mm × 165 mm × 95 mm) consists of four-outputs high-voltage power supply (HVPS) with maximum voltage of 3 KV and acquisition system (bipotentiostat) containing 2-channels for dual electrochemical detection. A new reusable microfluidic platform was designed in order to incorporate the microchips with the portable instrument. In this case, the platform is integrated to the main unit of the instrument so that it is not necessary to have any external cable for the interconnection of both parts, making the use of the complete system easier. The new platform contains all the electrical connections for the HVPS and bipotentiostat, as well as fluidic ports for driving the solutions. The microfluidic electrophoresis instrument is controlled by means of a user-friendly interface from a computer. The possibility of wireless connection (Bluetooth®) allows the use of the instrument without any external cable improving the portability. Therefore, the second generation brings a more compact and integrated electrophoresis instrument for "in situ" applications using microfluidic chips in an easy way. The performance of the electrophoresis system was initially evaluated using single- and dual-channel SU-8/Pyrex microchips with different models of integrated electrodes including microelectrodes and interdigitated arrays. The method was tested in different analytical applications such as separation of neurotransmitters, chlorophenols, purine derivatives, vitamins, polyphenolic acids, and flavones.

  8. A hybrid microfluidic chip with electrowetting functionality using ultraviolet (UV)-curable polymer.

    PubMed

    Gu, Hao; Duits, Michel H G; Mugele, Frieder

    2010-06-21

    Electrowetting (EW) is widely used in digital microfluidics for the manipulation of drops sandwiched between two parallel plates. In contrast, demonstrations of closed microfluidic channels enhanced with EW functionality are scarce. Here, we report a simple, low-cost method to construct such microchannels enclosed between two glass plates, each of which comprises electrodes and insulating layers. Our method uses soft imprint lithography with thiolene precursors to design the channel geometry. UV exposure is used to seal the chips permanently and a silanization treatment renders all inner channel surfaces hydrophobic. Compared to earlier polydimethylsiloxane-based designs, this method allows us to make microchannels with smaller dimensions (down to 10 microns), lower aspect ratios (down to height/length=1/10), and symmetric electrodes both on the top and the bottom of the channel. We demonstrate the new capabilities with two examples: (i) EW-enhanced drop generation in a flow focusing geometry allows precise and continuous control on drop diameter in the range approximately 1-15 microns while maintaining monodispersity; (ii) EW allows tuning of the excess water pressure needed to displace oil in a microchannel, leading to spontaneous imbibition at EW number eta>0.89.

  9. Single Cell Mass Measurement Using Drag Force Inside Lab-on-Chip Microfluidics System.

    PubMed

    Rahman, Md Habibur; Ahmad, Mohd Ridzuan; Takeuchi, Masaru; Nakajima, Masahiro; Hasegawa, Yasuhisa; Fukuda, Toshio

    2015-12-01

    Single cell mass (SCM) is an intrinsic property of single cell, it arouses a great interest among scientists as cell mass depends on the synthesis of proteins, DNA replication, cell wall stiffness, cell cytoplasm density, cell growth, ribosome, and other analogous of organisms. To date, several great strides have been taken to the advancements of SCM measurement techniques. Nevertheless, more works are required to enable the technology to push frontier in deep analysis of SCM measurement, hence to elucidate intracellular properties. In this paper, we present a lab-on-chip microfluidics system for SCM measurement, related with the force required to drag a single cell and Newton's law of motion inside microfluidics channel. Drag force on the cell was generated by a pressure driven syringe micropump and the motion of the cell was measured using optical observation under an inverted microscope. This approach of measuring SCM was calibrated using known mass (77.3 pg) of a polystyrene particle of 5.2 μm diameter. Furthermore, we used Saccharomyces cerevisiae baker's yeast cells of different sizes ([Formula: see text] diameter) for SCM measurement. Mass of 4.4 μm diameter of single yeast cell was measured as 2.12 pg which is in the range of previously reported single yeast cell mass (2-3 pg). In addition, we also studied the relation between SCM and single cell size. Results showed that single yeast cell mass increases exponentially with the increasing of single cell size.

  10. Single Cell Mass Measurement Using Drag ForceInside Lab-on-Chip Microfluidics System.

    PubMed

    Rahman, Md; Ahmad, Mohd; Takeuchi, Masaru; Nakajima, Masahiro; Hasegawa, Yasuhisa; Fukuda, Toshio

    2015-12-22

    Single Cell Mass (SCM) is an intrinsic property of single cell, it arouses a great interest among scientists as cell mass depends on the synthesis of proteins, DNA replication, cell wall stiffness, cell cytoplasm density, cell growth, ribosome and other analogous of organisms. To date, several great strides have been taken to the advancements of SCM measurement techniques. Nevertheless, more works are required to enable the technology to push frontier in deep analysis of SCM measurement, hence to elucidate intracellular properties. In this paper, we present a Lab-on-Chip microfluidics system for SCM measurement, related with the force required to drag a single cell and Newton's law of motion inside microfluidics channel. Drag force on the cell was generated by a pressure driven syringe micropump and the motion of the cell was measured using optical observation under an inverted microscope. This approach of measuring SCM was calibrated using known mass (77.3 pg) of a polystyrene particle of 5.2 μm diameter. Furthermore, we used Saccharomyces cerevisiae baker's yeast cells of different sizes (2-7 μm diameter) for SCM measurement. Mass of 4.4 μm diameter of single yeast cell was measured as 2.12 pg which is in the range of previously reported single yeast cell mass (2-3 pg). In addition, we also studied the relation between SCM and single cell size. Results showed that single yeast cell mass increases exponentially with the increasing of single cell size.

  11. On chip droplet characterization: a practical, high-sensitivity measurement of droplet impedance in digital microfluidics.

    PubMed

    Sadeghi, Saman; Ding, Huijiang; Shah, Gaurav J; Chen, Supin; Keng, Pei Yuin; Kim, Chang-Jin; van Dam, R Michael

    2012-02-21

    We demonstrate a new approach to impedance measurement on digital microfluidics chips for the purpose of simple, sensitive, and accurate volume and liquid composition measurement. Adding only a single series resistor to existing AC droplet actuation circuits, the platform is simple to implement and has negligible effect on actuation voltage. To accurately measure the complex voltage across the resistor (and hence current through the device and droplet), the designed system is based on software-implemented lock-in amplification detection of the voltage drop across the resistor which filters out noise, enabling high-resolution and low-limit signal recovery. We observe picoliter sensitivity with linear correlation of voltage to volume extending to the microliter volumes that can be handled by digital microfluidic devices. Due to the minimal hardware, the system is robust and measurements are highly repeatable. The detection technique provides both phase and magnitude information of the real-time current flowing through the droplet for a full impedance measurement. The sensitivity and resolution of this platform enables it to distinguish between various liquids which, as demonstrated in this paper, could potentially be extended to quantify solute concentrations, liquid mixtures, and presence of analytes.

  12. Recombinant spider silk from aqueous solutions via a bio-inspired microfluidic chip

    NASA Astrophysics Data System (ADS)

    Peng, Qingfa; Zhang, Yaopeng; Lu, Li; Shao, Huili; Qin, Kankan; Hu, Xuechao; Xia, Xiaoxia

    2016-11-01

    Spiders achieve superior silk fibres by controlling the molecular assembly of silk proteins and the hierarchical structure of fibres. However, current wet-spinning process for recombinant spidroins oversimplifies the natural spinning process. Here, water-soluble recombinant spider dragline silk protein (with a low molecular weight of 47 kDa) was adopted to prepare aqueous spinning dope. Artificial spider silks were spun via microfluidic wet-spinning, using a continuous post-spin drawing process (WS-PSD). By mimicking the natural spinning apparatus, shearing and elongational sections were integrated in the microfluidic spinning chip to induce assembly, orientation of spidroins, and fibril structure formation. The additional post-spin drawing process following the wet-spinning section partially mimics the spinning process of natural spider silk and substantially contributes to the compact aggregation of microfibrils. Subsequent post-stretching further improves the hierarchical structure of the fibres, including the crystalline structure, orientation, and fibril melting. The tensile strength and elongation of post-treated fibres reached up to 510 MPa and 15%, respectively.

  13. Recombinant spider silk from aqueous solutions via a bio-inspired microfluidic chip

    PubMed Central

    Peng, Qingfa; Zhang, Yaopeng; Lu, Li; Shao, Huili; Qin, Kankan; Hu, Xuechao; Xia, Xiaoxia

    2016-01-01

    Spiders achieve superior silk fibres by controlling the molecular assembly of silk proteins and the hierarchical structure of fibres. However, current wet-spinning process for recombinant spidroins oversimplifies the natural spinning process. Here, water-soluble recombinant spider dragline silk protein (with a low molecular weight of 47 kDa) was adopted to prepare aqueous spinning dope. Artificial spider silks were spun via microfluidic wet-spinning, using a continuous post-spin drawing process (WS-PSD). By mimicking the natural spinning apparatus, shearing and elongational sections were integrated in the microfluidic spinning chip to induce assembly, orientation of spidroins, and fibril structure formation. The additional post-spin drawing process following the wet-spinning section partially mimics the spinning process of natural spider silk and substantially contributes to the compact aggregation of microfibrils. Subsequent post-stretching further improves the hierarchical structure of the fibres, including the crystalline structure, orientation, and fibril melting. The tensile strength and elongation of post-treated fibres reached up to 510 MPa and 15%, respectively. PMID:27819339

  14. Microfluidic chip with optical sensor for rapid detection of nerve agent Sarin in water samples

    NASA Astrophysics Data System (ADS)

    Tan, Hsih Yin; Nguyen, Nam-Trung; Loke, Weng Keong; Tan, Yong Teng

    2007-12-01

    The chemical warfare agent Sarin is an organophosphate that is highly toxic to humans as they can act as cholinesterase inhibitors, that disrupts neuromuscular transmission. As these nerve agents are colorless, odorless and highly toxic, they can be introduced into drinking water as a means of terrorist sabotage. Hence, numerous innovative devices and methods have been developed for rapid detection of these organophosphates. Microfluidic technology allows the implementation of fast and sensitive detection of Sarin. In this paper, a micro-total analysis systems (TAS), also known as Lab-on-a-chip, fitted with an optical detection system has been developed to analyze the presence of the nerve agent sarin in water samples. In the present set-up, inhibition of co-introduced cholinesterase and water samples containing trace amounts of nerve agent sarin into the microfluidic device was used as the basis for selective detection of sarin. The device was fabricated using polymeric micromachining with PMMA (poly (methymethacrylate)) as the substrate material. A chromophore was utilized to measure the activity of remnant cholinesterase activity, which is inversely related to the amount of sarin present in the water samples. Comparisons were made between two different optical detection techniques and the findings will be presented in this paper. The presented measurement method is simple, fast and as sensitive as Gas Chromatography.

  15. Generation of Monodisperse Liquid Droplets in a Microfluidic Chip Using a High-Speed Gaseous Microflow

    NASA Astrophysics Data System (ADS)

    Tirandazi, Pooyan; Hidrovo, Carlos

    2015-11-01

    Over the last few years, microfluidic systems known as Lab-on-a-Chip (LOC) and micro total analysis systems (μTAS) have been increasingly developed as essential components for numerous biochemical applications. Droplet microfluidics, however, provides a distinctive attribute for delivering and processing discrete as well as ultrasmall volumes of fluid, which make droplet-based systems more beneficial over their continuous-phase counterparts. Droplet generation in its conventional scheme usually incorporates the injection of a liquid (water) into a continuous immiscible liquid (oil) medium. In this study we demonstrate a novel scheme for controlled generation of monodisperse droplets in confined gas-liquid microflows. We experimentally investigate the manipulation of water droplets in flow-focusing configurations using a high inertial air stream. Different flow regimes are observed by varying the gas and liquid flow rates, among which, the ``dripping regime'' where monodisperse droplets are generated is of great importance. The controlled size and generation rate of droplets in this region provide the capability for precise and contaminant-free delivery of microliter to nanoliter volumes of fluid. Furthermore, the high speed droplets generated in this method represent the basis for a new approach based on droplet pair collisions for fast efficient micromixing which provides a significant development in modern LOC and μTAS devices. This project is currently being supported by an NSF CAREER Award grant CBET-1151091.

  16. Combined Dielectrophoresis and Impedance Systems for Bacteria Analysis in Microfluidic On-Chip Platforms

    PubMed Central

    Páez-Avilés, Cristina; Juanola-Feliu, Esteve; Punter-Villagrasa, Jaime; del Moral Zamora, Beatriz; Homs-Corbera, Antoni; Colomer-Farrarons, Jordi; Miribel-Català, Pere Lluís; Samitier, Josep

    2016-01-01

    Bacteria concentration and detection is time-consuming in regular microbiology procedures aimed to facilitate the detection and analysis of these cells at very low concentrations. Traditional methods are effective but often require several days to complete. This scenario results in low bioanalytical and diagnostic methodologies with associated increased costs and complexity. In recent years, the exploitation of the intrinsic electrical properties of cells has emerged as an appealing alternative approach for concentrating and detecting bacteria. The combination of dielectrophoresis (DEP) and impedance analysis (IA) in microfluidic on-chip platforms could be key to develop rapid, accurate, portable, simple-to-use and cost-effective microfluidic devices with a promising impact in medicine, public health, agricultural, food control and environmental areas. The present document reviews recent DEP and IA combined approaches and the latest relevant improvements focusing on bacteria concentration and detection, including selectivity, sensitivity, detection time, and conductivity variation enhancements. Furthermore, this review analyses future trends and challenges which need to be addressed in order to successfully commercialize these platforms resulting in an adequate social return of public-funded investments. PMID:27649201

  17. Microfluidic devices for cell culture and handling in organ-on-a-chip applications

    NASA Astrophysics Data System (ADS)

    Becker, Holger; Schulz, Ingo; Mosig, Alexander; Jahn, Tobias; Gärtner, Claudia

    2014-03-01

    For many problems in system biology or pharmacology, in-vivo-like models of cell-cell interactions or organ functions are highly sought after. Conventional stationary cell culture in 2D plates quickly reaches its limitations with respect to an in-vivo like expression and function of individual cell types. Microfabrication technologies and microfluidics offer an attractive solution to these problems. The ability to generate flow as well as geometrical conditions for cell culture and manipulation close to the in-vivo situation allows for an improved design of experiments and the modeling of organ-like functionalities. Furthermore, reduced internal volumes lead to a reduction in reagent volumes necessary as well as an increased assay sensitivity. In this paper we present a range of microfluidic devices designed for the co-culturing of a variety of cells. The influence of substrate materials and surface chemistry on the cell morphology and viability for long-term cell culture has been investigated as well as strategies and medium supply for on-chip cell cultivation.

  18. Assessment of mitochondrial membrane potential using an on-chip microelectrode in a microfluidic device.

    PubMed

    Lim, Tae-Sun; Dávila, Antonio; Wallace, Douglas C; Burke, Peter

    2010-07-07

    The mitochondrial membrane potential is used to generate and regulate energy in living systems, driving the conversion of ADP to ATP, regulating ion homeostasis, and controlling apoptosis, all central to human health and disease. Therefore, there is a need for tools to study its regulation in a controlled environment for potential clinical and scientific applications. For this aim, an on-chip tetraphenylphosphonium (TPP(+)) selective microelectrode sensor was constructed in a microfluidic environment. The concentration of isolated mitochondria (Heb7A) used in a membrane potential measurement was 0.3 ng microL(-1), four orders of magnitude smaller than the concentration used in conventional assays (3 microg microL(-1)). In addition, the volume of the chamber (85 microL) is 2 orders of magnitude smaller than traditional experiments. As a demonstration, changes in the membrane potential are clearly measured in response to a barrage of well-known substrates and inhibitors of the electron transport chain. This general approach, which to date has not been demonstrated for study of mitochondrial function and bio-energetics in generally, can be instrumental in advancing the field of mitochondrial research and clinical applications by allowing high throughput studies of the regulation, dynamics, and statistical properties of the mitochondrial membrane potential in response to inhibitors and inducers of apoptosis in a controlled (microfluidic) chemical environment.

  19. High-throughput sorting of drops in microfluidic chips using electric capacitance

    PubMed Central

    Pit, Arjen M.; de Ruiter, Riëlle; Kumar, Anand; Wijnperlé, Daniel; Duits, Michèl H. G.; Mugele, Frieder

    2015-01-01

    We analyze a recently introduced approach for the sorting of aqueous drops with biological content immersed in oil, using a microfluidic chip that combines the functionality of electrowetting with the high throughput of two-phase flow microfluidics. In this electrostatic sorter, three co-planar electrodes covered by a thin dielectric layer are placed directly below the fluidic channel. Switching the potential of the central electrode creates an electrical guide that leads the drop to the desired outlet. The generated force, which deflects the drop, can be tuned via the voltage. The working principle is based on a contrast in conductivity between the drop and the continuous phase, which ensures successful operation even for drops of highly conductive biological media like phosphate buffered saline. Moreover, since the electric field does not penetrate the drop, its content is protected from electrical currents and Joule heating. A simple capacitive model allows quantitative prediction of the electrostatic forces exerted on drops. The maximum achievable sorting rate is determined by a competition between electrostatic and hydrodynamic forces. Sorting speeds up to 1200 per second are demonstrated for conductive drops of 160 pl in low viscosity oil. PMID:26339316

  20. Measurement of platelet aggregation functions using whole blood migration ratio in a microfluidic chip.

    PubMed

    Seo, Hong Seog; Choi, Sung Hyuk; Han, Miran; Kim, Kyeong Ah; Cho, Chi Hyun; An, Seong Soo A; Lim, Chae Seung; Shin, Sehyun

    2016-01-01

    Platelets play a major role in maintaining endothelial integrity and hemostasis. Of the various soluble agonists, ADP is an important in vivo stimulus for inducing platelet aggregation. In this study, a simple, rapid, and affordable method was designed for testing bleeding time (BT) and platelet aggregation with a two-channel microfluidic chip. Whole blood migration ratio (MR) from a microchip system was evaluated in comparison to the closure time (CT) from PFA-100 assays (Siemens, Germany) and CD62P expression on platelets. To induce platelet aggregation, a combination of collagen (1.84 mg/ml) and ADP (37.5 mg/ml) were used as agonists. After adding the agonists to samples, whole blood MR from the microchip system was measured. The outcome of the assessment depended on reaction time and agonist concentration. MR of whole blood from the microchip system was significantly correlated with CT from PFA-100 (r = 0.61, p <  0.05, n = 60). In addition, MR was negatively correlated with CD62P expression (r =-0.95, p <  0.05, n = 60). These results suggest that the measurement of MR using agonists is an easy, simple and efficient method for monitoring platelet aggregation in normal and ADP-receptors defective samples, along with the BT test. Thus, usage of the current microfluidic method could expand to diverse applications, including efficacy assessments in platelet therapy.

  1. Glucose microfluidic fuel cell based on silver bimetallic selective catalysts for on-chip applications

    NASA Astrophysics Data System (ADS)

    Cuevas-Muñiz, F. M.; Guerra-Balcázar, M.; Esquivel, J. P.; Sabaté, N.; Arriaga, L. G.; Ledesma-García, J.

    2012-10-01

    A glucose microfluidic fuel cell with outstanding performance at zero flow condition is presented. Polarization tests showed that bimetallic materials based in silver (AuAg/C as anode, PtAg/C as cathode) exhibit tolerance to byproducts and crossover effect. This allowed achieving one of the highest power densities reported for glucose fuel cells, up to a value of 630 μW cm-2 using two separated laminar flows of reactants. Furthermore, the tolerance to crossover effect caused by the selectivity of PtAg/C to oxygen reduction reaction in presence of glucose permitted using a single flow containing a mixture of glucose/oxygen, yielding a performance as high as 270 μW cm-2. Microfluidic fuel cell was further evaluated with a simulated body fluid solution that contained salts commonly present in the human blood plasma, reaching a power of 240 μW cm-2 at zero flow. These results envisage the incorporation of this fuel cell as a portable power source in Lab-on-a-Chip devices without the need of external pumps.

  2. Continuous enrichment of circulating tumor cells using a microfluidic lateral flow filtration chip.

    PubMed

    Lee, Sung-Woo; Hyun, Kyung-A; Kim, Seung-Il; Kang, Ji-Yoon; Jung, Hyo-Il

    2015-01-16

    The isolation and characterization of circulating tumor cells (CTC) is of great importance in cancer diagnosis and prognosis. Highly sensitive detection of CTCs can be very difficult because they are extremely rare (i.e., 1-5 CTCs per 10(9) erythrocytes) in blood. Recently, various devices have been developed that exploit biochemical (affinity-based) and physical (size or density) methods. Antibody-based isolation has its own limitations, as the expression level of the epitopes for an antibody varies due to the heterogeneity of cancer cells. Harsh conditions associated with physical methods can cause the deformation and damage of CTCs during the isolation process. Here, we propose a microfluidic lateral flow filtration (μ-LaFF) chip in which lateral flow was combined with vertical flow into the filter to capture the CTCs gently. The CTCs experienced weak shear flow owing to the lateral flow and traveled alongside the filter channel until finally being captured. The vertical flow in the filter held the captured cells tightly and served as an exit for uncaptured hematological cells (white and red blood cells). From our μ-LaFF chip we obtained a high capture efficiency (95%) and purity (99%), minimizing any damage to the CTCs. Our μ-LaFF technology is expected to be useful in the diagnosis and prognosis of various cancers.

  3. Rapid and Low-Cost CRP Measurement by Integrating a Paper-Based Microfluidic Immunoassay with Smartphone (CRP-Chip).

    PubMed

    Dong, Meili; Wu, Jiandong; Ma, Zimin; Peretz-Soroka, Hagit; Zhang, Michael; Komenda, Paul; Tangri, Navdeep; Liu, Yong; Rigatto, Claudio; Lin, Francis

    2017-03-26

    Traditional diagnostic tests for chronic diseases are expensive and require a specialized laboratory, therefore limiting their use for point-of-care (PoC) testing. To address this gap, we developed a method for rapid and low-cost C-reactive protein (CRP) detection from blood by integrating a paper-based microfluidic immunoassay with a smartphone (CRP-Chip). We chose CRP for this initial development because it is a strong biomarker of prognosis in chronic heart and kidney disease. The microfluidic immunoassay is realized by lateral flow and gold nanoparticle-based colorimetric detection of the target protein. The test image signal is acquired and analyzed using a commercial smartphone with an attached microlens and a 3D-printed chip-phone interface. The CRP-Chip was validated for detecting CRP in blood samples from chronic kidney disease patients and healthy subjects. The linear detection range of the CRP-Chip is up to 2 μg/mL and the detection limit is 54 ng/mL. The CRP-Chip test result yields high reproducibility and is consistent with the standard ELISA kit. A single CRP-Chip can perform the test in triplicate on a single chip within 15 min for less than 50 US cents of material cost. This CRP-Chip with attractive features of low-cost, fast test speed, and integrated easy operation with smartphones has the potential to enable future clinical PoC chronic disease diagnosis and risk stratification by parallel measurements of a panel of protein biomarkers.

  4. Multiscale variation-aware techniques for high-performance digital microfluidic lab-on-a-chip component placement.

    PubMed

    Liao, Chen; Hu, Shiyan

    2011-03-01

    The invention of microfluidic lab-on-a-chip alleviates the burden of traditional biochemical laboratory procedures which are often very expensive. Device miniaturization and increasing design complexity have mandated a shift in digital microfluidic lab-on-a-chip design from traditional manual design to computer-aided design (CAD) methodologies. As an important procedure in the lab-on-a-chip layout CAD, the lab-on-a-chip component placement determines the physical location and the starting time of each operation such that the overall completion time is minimized while satisfying nonoverlapping constraint, resource constraint, and scheduling constraint. In this paper, a multiscale variation-aware optimization technique based on integer linear programming is proposed for the lab-on-a-chip component placement. The simulation results demonstrate that without considering variations, our technique always satisfies the design constraints and largely outperforms the state-of-the-art approach, with up to 65.9% reduction in completion time. When considering variations, the variation-unaware design has the average yield of 2%, while our variation-aware technique always satisfies the yield constraint with only 7.7% completion time increase.

  5. Microfluidic chip with integrated electrical cell-impedance sensing for monitoring single cancer cell migration in three-dimensional matrixes.

    PubMed

    Nguyen, Tien Anh; Yin, Tsung-I; Reyes, Diego; Urban, Gerald A

    2013-11-19

    Cell migration has been recognized as one hallmark of malignant tumor progression. By integrating the method of electrical cell-substrate impedance sensing (ECIS) with the Boyden chamber design, the state-of-the-art techniques provide kinetic information about cell migration and invasion processes in three-dimensional (3D) extracellular matrixes. However, the information related to the initial stage of cell migration with single-cell resolution, which plays a unique role in the metastasis-invasion cascade of cancer, is not yet available. In this paper, we present a microfluidic device integrated with ECIS for investigating single cancer cell migration in 3D matrixes. Using microfluidics techniques without the requirement of physical connections to off-chip pneumatics, the proposed sensor chip can efficiently capture single cells on microelectrode arrays for sequential on-chip 2D or 3D cell culture and impedance measurement. An on-chip single-cell migration assay was successfully demonstrated within several minutes. Migration of single metastatic MDA-MB-231 cells in their initial stage can be monitored in real time; it shows a rapid change in impedance magnitude of approximately 10 Ω/s, whereas no prominent impedance change is observed for less-metastasis MCF-7 cells. The proposed sensor chip, allowing for a rapid and selective detection of the migratory properties of cancer cells at the single-cell level, could be applied as a new tool for cancer research.

  6. DNA-library assembly programmed by on-demand nano-liter droplets from a custom microfluidic chip

    PubMed Central

    Tangen, Uwe; Minero, Gabriel Antonio S.; Sharma, Abhishek; Wagler, Patrick F.; Cohen, Rafael; Raz, Ofir; Marx, Tzipy; Ben-Yehezkel, Tuval; McCaskill, John S.

    2015-01-01

    Nanoscale synthetic biology can benefit from programmable nanoliter-scale processing of DNA in microfluidic chips if they are interfaced effectively to biochemical arrays such as microwell plates. Whereas active microvalve chips require complex fabrication and operation, we show here how a passive and readily fabricated microchip can be employed for customizable nanoliter scale pipetting and reaction control involving DNA. This recently developed passive microfluidic device, supporting nanoliter scale combinatorial droplet generation and mixing, is here used to generate a DNA test library with one member per droplet exported to addressed locations on microwell plates. Standard DNA assembly techniques, such as Gibson assembly, compatible with isothermal on-chip operation, are employed and checked using off-chip PCR and assembly PCR. The control of output droplet sequences and mixing performance was verified using dyes and fluorescently labeled DNA solutions, both on-chip and in external capillary channels. Gel electrophoresis of products and DNA sequencing were employed to further verify controlled combination and functional enzymatic assembly. The scalability of the results to larger DNA libraries is also addressed by combinatorial input expansion using sequential injection plugs from a multiwell plate. Hence, the paper establishes a proof of principle of the production of functional combinatorial mixtures at the nanoliter scale for one sequence per well DNA libraries. PMID:26221198

  7. Processing window for femtosecond laser microsurgery and fluorescence imaging of an arterial tissue hosted in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Karimelahi, Samira; Li, Jianzhao; Herman, Peter R.

    2016-02-01

    We study the exposure limitations of femtosecond laser microsurgery and multiphoton imaging in a microfluidic chip environment, assessing damage thresholds at various interfaces as well as interference from bubble formation in the hosting solution. Both heat accumulation and incubation effects from multipulse laser exposures at 1-MHz repetition rate were evaluated. For demonstration, three microsurgery approaches of laser scribing, percussion drilling and trepanning were applied to arterial walls loaded in vitro in a lab-on-a-chip device. We report that deleterious effects from interface damage and microbubble formation can be avoided to offer laser processing windows for damage-free fluorescence imaging and precise microsurgery of live tissue hosted inside small microfluidic chambers.

  8. Continuous transfer of liquid metal droplets across a fluid-fluid interface within an integrated microfluidic chip.

    PubMed

    Gol, Berrak; Tovar-Lopez, Francisco J; Kurdzinski, Michael E; Tang, Shi-Yang; Petersen, Phred; Mitchell, Arnan; Khoshmanesh, Khashayar

    2015-06-07

    Micro scale liquid metal droplets have been hailed as the potential key building blocks of future micro-electro-mechanical systems (MEMS). However, most of the current liquid metal enabled systems involve millimeter scale droplets, which are manually injected onto the desired locations of the microchip. Despite its simplicity, this method is impractical for patterning large arrays or complex systems based on micro scale droplets. Here, we present a microfluidic chip, which integrates continuous generation of micro scale galinstan droplets in glycerol, and the hydrodynamic transfer of these droplets into sodium hydroxide (NaOH) solution. Observation via high-speed imaging along with computational fluid dynamics (CFD) analysis are utilised to comprehend the lateral migration of droplets from the glycerol to NaOH fluid. This platform is simple, can be readily integrated into other microfluidic systems, and creates flexibility by separating the continuous phase for droplet generation from the eventual target carrier fluid within a monolithic chip.

  9. A coral-on-a-chip microfluidic platform enabling live-imaging microscopy of reef-building corals.

    PubMed

    Shapiro, Orr H; Kramarsky-Winter, Esti; Gavish, Assaf R; Stocker, Roman; Vardi, Assaf

    2016-03-04

    Coral reefs, and the unique ecosystems they support, are facing severe threats by human activities and climate change. Our understanding of these threats is hampered by the lack of robust approaches for studying the micro-scale interactions between corals and their environment. Here we present an experimental platform, coral-on-a-chip, combining micropropagation and microfluidics to allow direct microscopic study of live coral polyps. The small and transparent coral micropropagates are ideally suited for live-imaging microscopy, while the microfluidic platform facilitates long-term visualization under controlled environmental conditions. We demonstrate the usefulness of this approach by imaging coral micropropagates at previously unattainable spatio-temporal resolutions, providing new insights into several micro-scale processes including coral calcification, coral-pathogen interaction and the loss of algal symbionts (coral bleaching). Coral-on-a-chip thus provides a powerful method for studying coral physiology in vivo at the micro-scale, opening new vistas in coral biology.

  10. On-Chip Microfluidic Components for In Situ Analysis, Separation, and Detection of Amino Acids

    NASA Technical Reports Server (NTRS)

    Zheng, Yun; Getty, Stephanie; Dworkin, Jason; Balvin, Manuel; Kotecki, Carl

    2013-01-01

    The Astrobiology Analytical Laboratory at GSFC has identified amino acids in meteorites and returned cometary samples by using liquid chromatography-electrospray ionization time-of-flight mass spectrometry (LCMS). These organic species are key markers for life, having the property of chirality that can be used to distinguish biological from non-biological amino acids. One of the critical components in the benchtop instrument is liquid chromatography (LC) analytical column. The commercial LC analytical column is an over- 250-mm-long and 4.6-mm-diameter stainless steel tube filled with functionized microbeads as stationary phase to separate the molecular species based on their chemistry. Miniaturization of this technique for spaceflight is compelling for future payloads for landed missions targeting astrobiology objectives. A commercial liquid chromatography analytical column consists of an inert cylindrical tube filled with a stationary phase, i.e., microbeads, that has been functionalized with a targeted chemistry. When analyte is sent through the column by a pressurized carrier fluid (typically a methanol/ water mixture), compounds are separated in time due to differences in chemical interactions with the stationary phase. Different species of analyte molecules will interact more strongly with the column chemistry, and will therefore take longer to traverse the column. In this way, the column will separate molecular species based on their chemistry. A lab-on-chip liquid analysis tool was developed. The microfluidic analytical column is capable of chromatographically separating biologically relevant classes of molecules based on their chemistry. For this analytical column, fabrication, low leak rate, and stationary phase incorporation of a serpentine microchannel were demonstrated that mimic the dimensions of a commercial LC column within a 5 10 1 mm chip. The microchannel in the chip has a 75- micrometer-diameter oval-shaped cross section. The serpentine

  11. A three dimensional thermoplastic microfluidic chip for robust cell capture and high resolution imaging

    PubMed Central

    Mottet, Guillaume; Perez-Toralla, Karla; Tulukcuoglu, Ezgi; Bidard, Francois-Clement; Pierga, Jean-Yves; Draskovic, Irena; Londono-Vallejo, Arturo; Descroix, Stephanie; Malaquin, Laurent; Louis Viovy, Jean

    2014-01-01

    We present a low cost microfluidic chip integrating 3D micro-chambers for the capture and the analysis of cells. This device has a simple design and a small footprint. It allows the implementation of standard biological protocols in a chip format with low volume consumption. The manufacturing process relies on hot-embossing of cyclo olefin copolymer, allowing the development of a low cost and robust device. A 3D design of microchannels was used to induce high flow velocity contrasts in the device and provide a selective immobilization. In narrow distribution channels, the liquid velocity induces a shear stress that overcomes adhesion forces and prevents cell immobilization or clogging. In large 3D chambers, the liquid velocity drops down below the threshold for cell attachment. The devices can be operated in a large range of input pressures and can even be handled manually using simple syringe or micropipette. Even at high flow injection rates, the 3D structures protect the captured cell from shear stress. To validate the performances of our device, we implemented immuno-fluorescence labeling and Fluorescence in Situ Hybridization (FISH) analysis on cancer cell lines and on a patient pleural effusion sample. FISH is a Food and Drug Administration approved cancer diagnostic technique that provides quantitative information about gene and chromosome aberration at the single cell level. It is usually considered as a long and fastidious test in medical diagnosis. This process can be easily implanted in our platform, and high resolution fluorescence imaging can be performed with reduced time and computer intensiveness. These results demonstrate the potential of this chip as a low cost, robust, and versatile tool adapted to complex and demanding protocols for medical diagnosis. PMID:25352942

  12. Adsorption and isolation of nucleic acids on cellulose magnetic beads using a three-dimensional printed microfluidic chip.

    PubMed

    Zhang, Lei; Deraney, Rachel N; Tripathi, Anubhav

    2015-11-01

    While advances in genomics have enabled sensitive and highly parallel detection of nucleic acid targets, the isolation and extraction of the nucleic acids remain a critical bottleneck in the workflow. We present here a simple 3D printed microfluidic chip that allows for the vortex and centrifugation free extraction of nucleic acids. This novel microfluidic chip utilizes the presence of a water and oil interface to filter out the lysate contaminants. The pure nucleic acids, while bound on cellulose particles, are magnetically moved across the oil layer. We demonstrated efficient and rapid extraction of spiked Human Papillomavirus (HPV) 18 plasmids in specimen transport medium, in under 15 min. An overall extraction efficiency of 61% is observed across a range of HPV plasmid concentrations (5 × 10(1) to 5 × 10(6) copies/100 μl). The magnetic, interfacial, and viscous drag forces inside the microgeometries of the chip are modeled. We have also developed a kinetics model for the adsorption of nucleic acids on cellulose functionalized superparamagnetic beads. We also clarify here the role of carrier nucleic acids in the adsorption and isolation of nucleic acids. Based on the various mechanistic insights detailed here, customized microfluidic devices can be designed to meet the range of current and emerging point of care diagnostics needs.

  13. A programmable and configurable multi-port System-on-Chip for stimulating electrokinetically-driven microfluidic devices.

    PubMed

    Lopez, Martha Salome; Gerstlauer, Andreas; Avila, Alfonso; Martinez-Chapa, Sergio O

    2011-01-01

    Recent research has demonstrated the use of microfluidic devices and electro-kinetics in areas such as medicine, genetics, embryology, epidemiology and pollution analysis, where manipulation of particles suspended in liquid media is required. Micro-fabrication technology has made it possible to increase system complexity and functionality by allowing integration of different processing and analysis stages in a single chip. However, fully integrated and autonomous microfluidic systems supporting ad-hoc stimulation have yet to be developed. This paper presents a flexible, configurable and programmable stimulator for electro-kinetically driven microfluidic devices. The stimulator is a dedicated System-on-Chip (SoC) architecture that generates sine, triangle, and sawtooth signals within a frequency range of 1 Hz to 20 MHz, capable of delivering single, dual, and superimposed waveforms, in a user defined test sequence for a selected time period. The system is designed to be integrated into complete, autonomous Lab-on-Chip, portable or implantable devices. As such, it is expected to help significantly advance current and future research on particle manipulation.

  14. Surface modification of PDMS microfluidic devices by controlled sulfuric acid treatment and the application in chip electrophoresis.

    PubMed

    Gitlin, Leonid; Schulze, Philipp; Ohla, Stefan; Bongard, Hans-Josef; Belder, Detlev

    2015-02-01

    Herein, we present a straightforward surface modification technique for PDMS-based microfluidic devices. The method takes advantage of the high reactivity of concentrated sulfuric acid to enhance the surface properties of PDMS bulk material. This results in alteration of the surface morphology and chemical composition that is in-depth characterized by ATR-FTIR, EDX, SEM, and XPS. In comparison to untreated PDMS, modified substrates exhibit a significantly reduced diffusive uptake of small organic molecules while retaining its low electroosmotic properties. This was demonstrated by exposing the channels of a microfluidic device to concentrated rhodamine B solution followed by fluorescence microscopy. The surface modification procedure was used to improve chip-based electrophoretic separations. Separation efficiencies of FITC-labeled amines/amino acids obtained in treated and untreated PDMS-devices as well as in glass chips were compared. We obtained higher efficiencies in H2 SO4 treated PDMS chips compared to untreated ones but lower efficiencies than those obtained in commercial microfluidic glass devices.

  15. Adsorption and isolation of nucleic acids on cellulose magnetic beads using a three-dimensional printed microfluidic chip

    PubMed Central

    Zhang, Lei; Deraney, Rachel N.; Tripathi, Anubhav

    2015-01-01

    While advances in genomics have enabled sensitive and highly parallel detection of nucleic acid targets, the isolation and extraction of the nucleic acids remain a critical bottleneck in the workflow. We present here a simple 3D printed microfluidic chip that allows for the vortex and centrifugation free extraction of nucleic acids. This novel microfluidic chip utilizes the presence of a water and oil interface to filter out the lysate contaminants. The pure nucleic acids, while bound on cellulose particles, are magnetically moved across the oil layer. We demonstrated efficient and rapid extraction of spiked Human Papillomavirus (HPV) 18 plasmids in specimen transport medium, in under 15 min. An overall extraction efficiency of 61% is observed across a range of HPV plasmid concentrations (5 × 101 to 5 × 106 copies/100 μl). The magnetic, interfacial, and viscous drag forces inside the microgeometries of the chip are modeled. We have also developed a kinetics model for the adsorption of nucleic acids on cellulose functionalized superparamagnetic beads. We also clarify here the role of carrier nucleic acids in the adsorption and isolation of nucleic acids. Based on the various mechanistic insights detailed here, customized microfluidic devices can be designed to meet the range of current and emerging point of care diagnostics needs. PMID:26734116

  16. A lab-on-a-chip system for the development of complex assays using modular microfluidic components

    NASA Astrophysics Data System (ADS)

    Hlawatsch, Nadine; Klemm, Richard; Carstens, Cornelia; Brandst"tter, Thomas; Becker, Holger; Elbracht, Rudi; Gärtner, Claudia

    2012-03-01

    For complex biological or diagnostic assays, the development of an integrated microfluidic device can be difficult and error-prone. For this reason, a modular approach, using individual microfluidic functional modules for the different process steps, can be advantageous. However often the interconnection of the modules proves to be tedious and the peripheral instrumentation to drive the various modules is cumbersome and of large size. For this reason, we have developed an integrated instrument platform which has generic functionalities such as valves and pumps, heating zones for continuous-flow PCR, moveable magnets for bead-based assays and an optical detection unit build into the instrument. The instrument holds a titerplate-sized carrier in which up to four microscopy-slide sized microfluidic modules can be clipped in. This allows for developing and optimizing individual assay steps without the need to modify the instrument or generate a completely new microfluidic cartridge. As a proof-of-concept, the automated sample processing of liquor or blood culture in microfluidic structures for detection of currently occuring Neisseria meningitidis strains was carried out. This assay involves the extraction of bacterial DNA, the fluorescent labeling, amplification using PCR as well as the hybridization of the DNA molecules in three-dimensional capture sites spotted into a microchannel. To define the assay sensitivity, chip modules were tested with bacteria spiked samples of different origins and results were controlled by conventional techniques. For liquor or blood culture, the presence of 200 bacteria was detected within 1 hour.

  17. Microfluidic and lab-on-a-chip preparation routes for organic nanoparticles and vesicular systems for nanomedicine applications.

    PubMed

    Capretto, Lorenzo; Carugo, Dario; Mazzitelli, Stefania; Nastruzzi, Claudio; Zhang, Xunli

    2013-11-01

    In recent years, advancements in the fields of microfluidic and lab-on-a-chip technologies have provided unique opportunities for the implementation of nanomaterial production processes owing to the miniaturisation of the fluidic environment. It has been demonstrated that microfluidic reactors offer a range of advantages compared to conventional batch reactors, including improved controllability and uniformity of nanomaterial characteristics. In addition, the fast mixing achieved within microchannels, and the predictability of the laminar flow conditions, can be leveraged to investigate the nanomaterial formation dynamics. In this article recent developments in the field of microfluidic production of nanomaterials for drug delivery applications are reviewed. The features that make microfluidic reactors a suitable technological platform are discussed in terms of controllability of nanomaterials production. An overview of the various strategies developed for the production of organic nanoparticles and colloidal assemblies is presented, focusing on those nanomaterials that could have an impact on nanomedicine field such as drug nanoparticles, polymeric micelles, liposomes, polymersomes, polyplexes and hybrid nanoparticles. The effect of microfluidic environment on nanomaterials formation dynamics, as well as the use of microdevices as tools for nanomaterial investigation is also discussed.

  18. A rapid and sensitive method for hydroxyl radical detection on a microfluidic chip using an N-doped porous carbon nanofiber modified pencil graphite electrode.

    PubMed

    Ouyang, Jun; Li, Zhong-Qiu; Zhang, Jing; Wang, Chen; Wang, Jiong; Xia, Xing-Hua; Zhou, Guo-Jun

    2014-07-07

    Hydroxyl radicals (˙OH) play an important role in human diseases. Traditional detection methods are time consuming and require expensive instruments. Here, we present a simple and sensitive method for the detection of hydroxyl radicals on a microfluidic chip using an electrochemical technique. Aniline monomer is electrochemically polymerized on the surface of a pencil graphite electrode and carbonized at 800 °C. The resulting N-doped porous carbon nanofiber-modified pencil graphite electrode is embedded into a microfluidic chip directly as a working electrode. 4-Hydroxybenzoic acid (4-HBA) is selected as the trapping agent owing to its unique 3,4-DHBA product and high trapping efficiency. A low detection limit of 1.0 × 10(-6) M is achieved on the microfluidic chip. As a demonstration, the microfluidic chip is successfully utilized for the detection of ˙OH in cigarette smoke. The strong π-π stacking and hydrophobic interactions between the nitrogen-doped carbon materials and the pencil graphite make the modified electrode well-suited for the microfluidic chip.

  19. Beating heart on a chip: a novel microfluidic platform to generate functional 3D cardiac microtissues.

    PubMed

    Marsano, Anna; Conficconi, Chiara; Lemme, Marta; Occhetta, Paola; Gaudiello, Emanuele; Votta, Emiliano; Cerino, Giulia; Redaelli, Alberto; Rasponi, Marco

    2016-02-07

    In the past few years, microfluidic-based technology has developed microscale models recapitulating key physical and biological cues typical of the native myocardium. However, the application of controlled physiological uniaxial cyclic strains on a defined three-dimension cellular environment is not yet possible. Two-dimension mechanical stimulation was particularly investigated, neglecting the complex three-dimensional cell-cell and cell-matrix interactions. For this purpose, we developed a heart-on-a-chip platform, which recapitulates the physiologic mechanical environment experienced by cells in the native myocardium. The device includes an array of hanging posts to confine cell-laden gels, and a pneumatic actuation system to induce homogeneous uniaxial cyclic strains to the 3D cell constructs during culture. The device was used to generate mature and highly functional micro-engineered cardiac tissues (μECTs), from both neonatal rat and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM), strongly suggesting the robustness of our engineered cardiac micro-niche. Our results demonstrated that the cyclic strain was effectively highly uniaxial and uniformly transferred to cells in culture. As compared to control, stimulated μECTs showed superior cardiac differentiation, as well as electrical and mechanical coupling, owing to a remarkable increase in junction complexes. Mechanical stimulation also promoted early spontaneous synchronous beating and better contractile capability in response to electric pacing. Pacing analyses of hiPSC-CM constructs upon controlled administration of isoprenaline showed further promising applications of our platform in drug discovery, delivery and toxicology fields. The proposed heart-on-a-chip device represents a relevant step forward in the field, providing a standard functional three-dimensional cardiac model to possibly predict signs of hypertrophic changes in cardiac phenotype by mechanical and biochemical co-stimulation.

  20. Chemical stimulation of the Arabidopsis thaliana root using multi-laminar flow on a microfluidic chip.

    PubMed

    Meier, Matthias; Lucchetta, Elena M; Ismagilov, Rustem F

    2010-08-21

    In this article, we developed a "plant on a chip" microfluidic platform that can control the local chemical environment around live roots of Arabidopsis thaliana with high spatial resolution using multi-laminar flow. We characterized the flow profile around the Arabidopsis root, and verified that the shear forces within the device ( approximately 10 dyne cm(-2)) did not impede growth of the roots. Our platform was able to deliver stimuli to the root at a spatial resolution of 10-800 microm. Further, the platform was validated by exposing desired regions of the root with a synthetic auxin derivative, 2,4-dichlorophenoxyacetic acid (2,4-D), and its inhibitor N-1-naphthylphthalamic acid (NPA). The response to the stimuli was observed using a DR5::GFP Arabidopsis line, where GFP expression is coupled to the auxin response regulator DR5. GFP expression in the root matched the position of the flow-focused stream containing 2,4-D. When the regions around the 2,4-D stimulus were exposed to the auxin transport inhibitor NPA, the active and passive transport mechanisms of auxin could be differentiated, as NPA blocks active cell-to-cell transport of auxin. Finally, we demonstrated that local 2,4-D stimulation in a approximately 10 microm root segment enhanced morphological changes such as epidermal hair growth. These experiments were proof-of-concept and agreed with the results expected based on known root biology, demonstrating that this "root on a chip" platform can be used to test how root development is affected by any chemical component of interest, including nitrogen, phosphate, salts, and other plant hormones.

  1. Gradient Static-Strain Stimulation in a Microfluidic Chip for 3D Cellular Alignment

    PubMed Central

    Hsieh, Hsin-Yi; Camci-Unal, Gulden; Huang, Tsu-Wei; Liao, Ronglih; Chen, Tsung-Ju; Paul, Arghya; Tseng, Fan-Gang; Khademhosseini, Ali

    2014-01-01

    Cell alignment is a critical factor to govern cellular behavior and function for various tissue engineering applications ranging from cardiac to neural regeneration. In addition to physical geometry, strain is a crucial parameter to manipulate cellular alignment for functional tissue formation. In this paper, we introduce a simple approach to generate a range of gradient static strains without external mechanical control for the stimulation of cellular behavior within 3D biomimetic hydrogel microenvironments. A glass-supported microfluidic chip with a convex flexible polydimethylsiloxane (PDMS) membrane on the top was employed for loading the cells suspended in a prepolymer solution. Following UV crosslinking through a photomask with a concentric circular pattern, the cell-laden hydrogels were formed in a height gradient from the center (maximum) to the boundary (minimum). When the convex PDMS membrane retracted back to a flat surface, it applied compressive gradient forces on the cell-laden hydrogels. The concentric circular hydrogel patterns confined the direction of hydrogel elongation, and the compressive strain on the hydrogel therefore resulted in elongation stretch in the radial direction to guide cell alignment. NIH3T3 cells were cultured in the chip for 3 days with compressive strains that varied from ~65% (center) to ~15% (boundary) on hydrogels. We found that the hydrogel geometry dominated the cell alignment near the outside boundary, where cells aligned along the circular direction, and the compressive strain dominated the cell alignment near the center, where cells aligned radially. This study developed a new and simple approach to facilitate cellular alignment based on hydrogel geometry and strain stimulation for tissue engineering applications. This platform offers unique advantages and is significantly different than the existing approaches owing to the fact that gradient generation was accomplished in a miniature device without using an external

  2. NeuroChip: A Microfluidic Electrophysiological Device for Genetic and Chemical Biology Screening of Caenorhabditis elegans Adult and Larvae

    PubMed Central

    Hu, Chunxiao; Dillon, James; Kearn, James; Murray, Caitriona; O’Connor, Vincent; Holden-Dye, Lindy; Morgan, Hywel

    2013-01-01

    Genetic and chemical biology screens of C. elegans have been of enormous benefit in providing fundamental insight into neural function and neuroactive drugs. Recently the exploitation of microfluidic devices has added greater power to this experimental approach providing more discrete and higher throughput phenotypic analysis of neural systems. Here we make a significant addition to this repertoire through the design of a semi-automated microfluidic device, NeuroChip, which has been optimised for selecting worms based on the electrophysiological features of the pharyngeal neural network. We demonstrate this device has the capability to sort mutant from wild-type worms based on high definition extracellular electrophysiological recordings. NeuroChip resolves discrete differences in excitatory, inhibitory and neuromodulatory components of the neural network from individual animals. Worms may be fed into the device consecutively from a reservoir and recovered unharmed. It combines microfluidics with integrated electrode recording for sequential trapping, restraining, recording, releasing and recovering of C. elegans. Thus mutant worms may be selected, recovered and propagated enabling mutagenesis screens based on an electrophysiological phenotype. Drugs may be rapidly applied during the recording thus permitting compound screening. For toxicology, this analysis can provide a precise description of sub-lethal effects on neural function. The chamber has been modified to accommodate L2 larval stages showing applicability for small size nematodes including parasitic species which otherwise are not tractable to this experimental approach. We also combine NeuroChip with optogenetics for targeted interrogation of the function of the neural circuit. NeuroChip thus adds a new tool for exploitation of C. elegans and has applications in neurogenetics, drug discovery and neurotoxicology. PMID:23717588

  3. Development of micropump-actuated negative pressure pinched injection for parallel electrophoresis on array microfluidic chip.

    PubMed

    Li, Bowei; Jiang, Lei; Xie, Hua; Gao, Yan; Qin, Jianhua; Lin, Bingcheng

    2009-09-01

    A micropump-actuated negative pressure pinched injection method is developed for parallel electrophoresis on a multi-channel LIF detection system. The system has a home-made device that could individually control 16-port solenoid valves and a high-voltage power supply. The laser beam is excitated and distributes to the array separation channels for detection. The hybrid Glass-PDMS microfluidic chip comprises two common reservoirs, four separation channels coupled to their respective pneumatic micropumps and two reference channels. Due to use of pressure as a driving force, the proposed method has no sample bias effect for separation. There is only one high-voltage supply needed for separation without relying on the number of channels, which is significant for high-throughput analysis, and the time for sample loading is shortened to 1 s. In addition, the integrated micropumps can provide the versatile interface for coupling with other function units to satisfy the complicated demands. The performance is verified by separation of DNA marker and Hepatitis B virus DNA samples. And this method is also expected to show the potential throughput for the DNA analysis in the field of disease diagnosis.

  4. Chitosan microgels obtained by on-chip crosslinking reaction employing a microfluidic device

    NASA Astrophysics Data System (ADS)

    Zamora-Mora, Vanessa; Velasco, Diego; Hernández, Rebeca; Mijangos, Carmen

    2014-12-01

    In the present work, we report on the preparation of microgels of chitosan crosslinked with sodium tripolyphosphate (TPP) employing the microfluidics technique (MF). To achieve this, several flow focusing geometries were designed and tested. As a first step, a two-inlet flow focusing geometry was employed to emulsify chitosan and the crosslinking reaction was carried out offchip. This procedure did not allow separating the resulting chitosan microgels due to an incomplete crosslinking reaction. A crosslinking reaction on-chip was studied as an alternative. A four-inlet flow focusing geometrywas designed in which three dispersed phases, chitosan 0.25% (w/v), TPP 0.05% (w/v) and acetic acid 1% (v/v) and an continuous phase mineral oil + Span 80 (3% w/v) were employed. The flow rates for the continuous phase were varied from 6.7 to 11.7 μL/min and chitosan microgels were successfully obtained with average diameters from 68 to 42 μm. The average size of the microgels outside the MF device decreased up to ~21% with respect to their size inside the MF device due to partial expulsion of water from the microgels when complete gelation occurred.

  5. Real-time machine vision FPGA implementation for microfluidic monitoring on Lab-on-Chips.

    PubMed

    Sotiropoulou, Calliope-Louisa; Voudouris, Liberis; Gentsos, Christos; Demiris, Athanasios M; Vassiliadis, Nikolaos; Nikolaidis, Spyridon

    2014-04-01

    A machine vision implementation on a field-programmable gate array (FPGA) device for real-time microfluidic monitoring on Lab-On-Chips is presented in this paper. The machine vision system is designed to follow continuous or plug flows, for which the menisci of the fluids are always visible. The system discriminates between the front or "head" of the flow and the back or "tail" and is able to follow flows with a maximum speed of 20 mm/sec in circular channels of a diameter of 200 μm (corresponding to approx. 60 μl/sec ). It is designed to be part of a complete Point-of-Care system, which will be portable and operate in non-ideal laboratory conditions. Thus, it is able to cope with noise due to lighting conditions and small LoC displacements during the experiment execution. The machine vision system can be used for a variety of LoC devices, without the need for fiducial markers (such as redundancy patterns) for its operation. The underlying application requirements called for a complete hardware implementation. The architecture uses a variety of techniques to improve performance and minimize memory access requirements. The system input is 8 bit grayscale uncompressed video of up to 1 Mpixel resolution. The system uses an operating frequency of 170 Mhz and achieves a computational time of 13.97 ms (worst case), which leads to a throughput of 71.6 fps for 1 Mpixel video resolution.

  6. Isolation of motile spermatozoa with a microfluidic chip having a surface-modified microchannel.

    PubMed

    Huang, Hong-Yuan; Wu, Tsung-Lin; Huang, Hung-Ru; Li, Chin-Jung; Fu, Hui-Ting; Soong, Yung-Kuei; Lee, Ming-Yih; Yao, Da-Jeng

    2014-02-01

    Conventional methods to prepare sperm have been amenable to the investigation of outcomes such as rates of recovery and conventional semen parameters. The standard preparation of sperm for assisted reproduction is criticized for its centrifugation steps, which might either recover motile sperm in variable proportions or increase the probability of damage to sperm DNA. An microfluidic system was designed to separate motile sperm according to a design whereby nonmotile spermatozoa and debris flow along their initial streamlines and exit through one outlet-up, whereas motile spermatozoa have an opportunity to swim into a parallel stream and to exit through a separate outlet-down. This chip was fabricated by microelectromechanical systems technology with polydimethylsiloxane molding. The hydrophilic surface, coated with poly (ethanediol) methyl ether methacrylate, exhibits enduring stability maintained for the microchannel. Microscopic examination and fluorescent images showed that the motility of sperm varied with the laminar streams. To confirm the sorting, we identified and quantified the proportions of live and dead sperm before and after sorting with flow cytometric analysis. The results on the viability of a sample demonstrated the increased quality of sperm after sorting and collection in the outlet reservoir. The counted ratio of live sperm revealed the quantity and efficiency of the sorted sperm.

  7. An integrated microfluidic chip system for single-cell secretion profiling of rare circulating tumor cells.

    PubMed

    Deng, Yuliang; Zhang, Yu; Sun, Shuai; Wang, Zhihua; Wang, Minjiao; Yu, Beiqin; Czajkowsky, Daniel M; Liu, Bingya; Li, Yan; Wei, Wei; Shi, Qihui

    2014-12-16

    Genetic and transcriptional profiling, as well as surface marker identification of single circulating tumor cells (CTCs) have been demonstrated. However, quantitatively profiling of functional proteins at single CTC resolution has not yet been achieved, owing to the limited purity of the isolated CTC populations and a lack of single-cell proteomic approaches to handle and analyze rare CTCs. Here, we develop an integrated microfluidic system specifically designed for streamlining isolation, purification and single-cell secretomic profiling of CTCs from whole blood. Key to this platform is the use of photocleavable ssDNA-encoded antibody conjugates to enable a highly purified CTC population with <75 'contaminated' blood cells. An enhanced poly-L-lysine barcode pattern is created on the single-cell barcode chip for efficient capture rare CTC cells in microchambers for subsequent secreted protein profiling. This system was extensively evaluated and optimized with EpCAM-positive HCT116 cells seeded into whole blood. Patient blood samples were employed to assess the utility of the system for isolation, purification and single-cell secretion profiling of CTCs. The CTCs present in patient blood samples exhibit highly heterogeneous secretion profile of IL-8 and VEGF. The numbers of secreting CTCs are found not in accordance with CTC enumeration based on immunostaining in the parallel experiments.

  8. Construction and operation of a microrobot based on magnetotactic bacteria in a microfluidic chip

    PubMed Central

    Ma, Qiufeng; Chen, Changyou; Wei, Shufeng; Chen, Chuanfang; Wu, Long-Fei; Song, Tao

    2012-01-01

    Magnetotactic bacteria (MTB) are capable of swimming along magnetic field lines. This unique feature renders them suitable in the development of magnetic-guided, auto-propelled microrobots to serve in target molecule separation and detection, drug delivery, or target cell screening in a microfluidic chip. The biotechnology to couple these bacteria with functional loads to form microrobots is the critical point in its application. Although an immunoreaction approach to attach functional loads to intact MTB was suggested, details on its realization were hardly mentioned. In the current paper, MTB-microrobots were constructed by attaching 2 μm diameter microbeads to marine magnetotactic ovoid MO-1 cells through immunoreactions. These microrobots were controlled using a special control and tracking system. Experimental results prove that the attachment efficiency can be improved to ∼30% via an immunoreaction. The motility of the bacteria attached with different number of loads was also assessed. The results show that MTB can transport one load at a velocity of ∼21 μm/s and still move and survive for over 30 min. The control and tracking system is fully capable of directing and monitoring the movement of the MTB-microrobots. The rotating magnetic fields can stop the microrobots by trapping them as they swim within a circular field with a controllable size. The system has potential use in chemical analyses and medical diagnoses using biochips as well as in nano/microscale transport. PMID:22655018

  9. The Optimization of Electrophoresis on a Glass Microfluidic Chip and its Application in Forensic Science.

    PubMed

    Han, Jun P; Sun, Jing; Wang, Le; Liu, Peng; Zhuang, Bin; Zhao, Lei; Liu, Yao; Li, Cai X

    2017-02-07

    Microfluidic chips offer significant speed, cost, and sensitivity advantages, but numerous parameters must be optimized to provide microchip electrophoresis detection. Experiments were conducted to study the factors, including sieving matrices (the concentration and type), surface modification, analysis temperature, and electric field strengths, which all impact the effectiveness of microchip electrophoresis detection of DNA samples. Our results showed that the best resolution for ssDNA was observed using 4.5% w/v (7 M urea) lab-fabricated LPA gel, dynamic wall coating of the microchannel, electrophoresis temperatures between 55 and 60°C, and electrical fields between 350 and 450 V/cm on the microchip-based capillary electrophoresis (μCE) system. One base-pair resolution could be achieved in the 19-cm-length microchannel. Furthermore, both 9947A standard genomic DNA and DNA extracted from blood spots were demonstrated to be successfully separated with well-resolved DNA peaks in 8 min. Therefore, the microchip electrophoresis system demonstrated good potential for rapid forensic DNA analysis.

  10. A controlled microfluidic electrochemical lab-on-a-chip for label-free diffusion-restricted DNA hybridization analysis.

    PubMed

    Ben-Yoav, Hadar; Dykstra, Peter H; Bentley, William E; Ghodssi, Reza

    2015-02-15

    Lab-on-a-chip (LOC) devices for electrochemical analysis of DNA hybridization events offer a technology for real-time and label-free assessment of biomarkers at the point-of-care. Here, we present a microfluidic LOC, with 3 × 3 arrayed electrochemical sensors for the analysis of DNA hybridization events. A new dual layer microfluidic valved manipulation system is integrated providing controlled and automated capabilities for high throughput analysis. This feature improves the repeatability, accuracy, and overall sensing performance (Fig. 1). The electrochemical activity of the fabricated microfluidic device is validated and demonstrated repeatable and reversible Nernstian characteristics. System design required detailed analysis of energy storage and dissipation as our sensing modeling involves diffusion-related electrochemical impedance spectroscopy. The effect of DNA hybridization on the calculated charge transfer resistance and the diffusional resistance components is evaluated. We demonstrate a specific device with an average cross-reactivity value of 27.5%. The device yields semilogarithmic dose response and enables a theoretical detection limit of 1 nM of complementary ssDNA target. This limit is lower than our previously reported non-valved device by 74% due to on-chip valve integration providing controlled and accurate assay capabilities.

  11. An Aluminum Microfluidic Chip Fabrication Using a Convenient Micromilling Process for Fluorescent Poly(dl-lactide-co-glycolide) Microparticle Generation

    PubMed Central

    Lin, Yung-Sheng; Yang, Chih-Hui; Wang, Chih-Yu; Chang, Fang-Rong; Huang, Keng-Shiang; Hsieh, Wan-Chen

    2012-01-01

    This study presents the development of a robust aluminum-based microfluidic chip fabricated by conventional mechanical micromachining (computer numerical control-based micro-milling process). It applied the aluminum-based microfluidic chip to form poly(lactic-co-glycolic acid) (PLGA) microparticles encapsulating CdSe/ZnS quantum dots (QDs). A cross-flow design and flow-focusing system were employed to control the oil-in-water (o/w) emulsification to ensure the generation of uniformly-sized droplets. The size of the droplets could be tuned by adjusting the flow rates of the water and oil phases. The proposed microfluidic platform is easy to fabricate, set up, organize as well as program, and is valuable for further applications under harsh reaction conditions (high temperature and/or strong organic solvent systems). The proposed method has the advantages of actively controlling the droplet diameter, with a narrow size distribution, good sphericity, as well as being a simple process with a high throughput. In addition to the fluorescent PLGA microparticles in this study, this approach can also be applied to many applications in the pharmaceutical and biomedical area. PMID:22438719

  12. Epithelial cell adhesion molecule independent capture of non-small lung carcinoma cells with peptide modified microfluidic chip.

    PubMed

    Pu, Kefeng; Li, Chunlin; Zhang, Nengpan; Wang, Hui; Shen, Wenjiang; Zhu, Yimin

    2017-03-15

    Circulating tumor cells (CTCs) present in the blood of patients with non-hematological cancers are accessible sources for diagnosis and monitoring of cancers. By the aid of the ability of the anti-EpCAM antibody to recognize the epithelial cells, microsystem-based technologies provide robust means for effectively detecting CTCs in vitro. Considering the EpCAM expression is down-regulated during epithelial-mesenchymal transition (EMT) process, the amount of CTCs detected based on anti-EpCAM antibody is underestimated. In our study, the A549 cells targeting peptide (A-1 peptide), as the substitute of anti-EpCAM antibody, was introduced to microfluidic chip to capture A549 cells. Our results showed that both epithelial-like and mesenchymal-like A549 cells could efficiently be captured by the A-1 peptide modified microfluidic chip, and the capture efficiency for epithelial-like cells is comparable to that captured by the EpCAM antibody. Thus, we concluded that the peptide could be a better supplement to the EpCAM antibody for capturing CTCs in microfluidic system with broader spectrum.

  13. Microfluidics and photonics for Bio-System-on-a-Chip: a review of advancements in technology towards a microfluidic flow cytometry chip.

    PubMed

    Godin, Jessica; Chen, Chun-Hao; Cho, Sung Hwan; Qiao, Wen; Tsai, Frank; Lo, Yu-Hwa

    2008-10-01

    Microfluidics and photonics come together to form a field commonly referred to as 'optofluidics'. Flow cytometry provides the field with a technology base from which both microfluidic and photonic components be developed and integrated into a useful device. This article reviews some of the more recent developments to familiarize a reader with the current state of the technologies and also highlights the requirements of the device and how researchers are working to meet these needs.

  14. Bioprinting cell-laden matrigel for radioprotection study of liver by pro-drug conversion in a dual-tissue microfluidic chip.

    PubMed

    Snyder, J E; Hamid, Q; Wang, C; Chang, R; Emami, K; Wu, H; Sun, W

    2011-09-01

    The objective of this paper is to introduce a novel cell printing and microfluidic system to serve as a portable ground model for the study of drug conversion and radiation protection of living liver tissue analogs. The system is applied to study behavior in ground models of space stress, particularly radiation. A microfluidic environment is engineered by two cell types to prepare an improved higher fidelity in vitro micro-liver tissue analog. Cell-laden Matrigel printing and microfluidic chips were used to test radiation shielding to liver cells by the pro-drug amifostine. In this work, the sealed microfluidic chip regulates three variables of interest: radiation exposure, anti-radiation drug treatment and single- or dual-tissue culture environments. This application is intended to obtain a scientific understanding of the response of the multi-cellular biological system for long-term manned space exploration, disease models and biosensors.

  15. Selecting and designing with the right thermoplastic polymer for your microfluidic chip: a close look into cyclo-olefin polymer

    NASA Astrophysics Data System (ADS)

    Nevitt, Mark

    2013-03-01

    Engineers who are developing microfluidic devices and bioMEMs for life science applications have many aspects to consider when selecting the proper base materials for constructing a device. While glass and polydimethylsiloxane (PDMS) are the staple materials for proof-of-concept and prototype chip fabrication, they are not a feasible solution for commercial production due to their slow, labor-intensive production rate. Alternatively, a molded or extruded thermoplastic solution can deliver the precision, consistency, and high volume capability required for commercial scale production. Traditional thermoplastics, such as polymethylmethacrylate (PMMA), polycarbonate (PC), and polystyrene (PS), are well known by development engineers in the bioscience community; however, cyclo-olefin polymer (COP), a relative newcomer in the world of plastics, is gaining increasing attention for use in microfluidic devices due to its unique balance of key properties compared to conventional thermoplastics. In this paper, we provide a comprehensive look at the properties which make COP an excellent candidate for providing the flow cell support and reagent storage functions in microfluidic assays. We also explore the processing attributes and capabilities of COP resin and film which are crucial for manufacturing high-performance microfluidic devices.

  16. Dopamine-functionalized InP/ZnS quantum dots as fluorescence probes for the detection of adenosine in microfluidic chip

    PubMed Central

    Ankireddy, Seshadri Reddy; Kim, Jongsung

    2015-01-01

    Microbeads are frequently used as solid supports for biomolecules such as proteins and nucleic acids in heterogeneous microfluidic assays. Chip-based, quantum dot (QD)-bead-biomolecule probes have been used for the detection of various types of DNA. In this study, we developed dopamine (DA)-functionalized InP/ZnS QDs (QDs-DA) as fluorescence probes for the detection of adenosine in microfluidic chips. The photoluminescence (PL) intensity of the QDs-DA is quenched by Zn2+ because of the strong coordination interactions. In the presence of adenosine, Zn2+ cations preferentially bind to adenosine, and the PL intensity of the QDs-DA is recovered. A polydimethylsiloxane-based microfluidic chip was fabricated, and adenosine detection was confirmed using QDs-DA probes. PMID:26347351

  17. Real-Time Electrical Impedimetric Monitoring of Blood Coagulation Process under Temperature and Hematocrit Variations Conducted in a Microfluidic Chip

    PubMed Central

    Lei, Kin Fong; Chen, Kuan-Hao; Tsui, Po-Hsiang; Tsang, Ngan-Ming

    2013-01-01

    Blood coagulation is an extremely complicated and dynamic physiological process. Monitoring of blood coagulation is essential to predict the risk of hemorrhage and thrombosis during cardiac surgical procedures. In this study, a high throughput microfluidic chip has been developed for the investigation of the blood coagulation process under temperature and hematocrit variations. Electrical impedance of the whole blood was continuously recorded by on-chip electrodes in contact with the blood sample during coagulation. Analysis of the impedance change of the blood was conducted to investigate the characteristics of blood coagulation process and the starting time of blood coagulation was defined. The study of blood coagulation time under temperature and hematocrit variations was shown a good agreement with results in the previous clinical reports. The electrical impedance measurement for the definition of blood coagulation process provides a fast and easy measurement technique. The microfluidic chip was shown to be a sensitive and promising device for monitoring blood coagulation process even in a variety of conditions. It is found valuable for the development of point-of-care coagulation testing devices that utilizes whole blood sample in microliter quantity. PMID:24116099

  18. Fabrication of high-aspect-ratio microstructures in polymer microfluid chips for in vitro single-cell analysis

    NASA Astrophysics Data System (ADS)

    Bukatin, A. S.; Mukhin, I. S.; Malyshev, E. I.; Kukhtevich, I. V.; Evstrapov, A. A.; Dubina, M. V.

    2016-10-01

    Technologies and methods of prototyping microfluidic devices are widely used in solving many biological problems and testing of operability of new microanalytic systems. This study is devoted to analyzing the features of the formation of microstructures in SU-8 photoresist and the preparation of replicas in polydimethyl siloxane by the soft lithography method. It has been shown that the aspect ratio of the resultant microstructures is determined by their shape, size, and the force of resist adhesion to the silicon substrate and the efficiency of the circulation of the developer around microstructures. In the replication of complex microstructures, an aspect ratio of 25 is attained. The technology considered here is used to prepare microfluidic chips with mechanical traps for fixation and the in vitro analysis of living cells.

  19. Imaging new transient nanostructures using a microfluidic chip integrated with a controlled environment vitrification system for cryogenic transmission electron microscopy.

    PubMed

    Lee, Jinkee; Jha, Ashish K; Bose, Arijit; Tripathi, Anubhav

    2008-11-18

    Nanostructures (vesicles, micelles, bilayers) are important in nanomedicine and biochemical processes. They are agents for encapsulation and eventual release of drugs, flavors, and fragrances. The structural transition from micelles to vesicles through disk-like intermediate states has been demonstrated previously. Here, we disclose a new route for the micelle-vesicle transition, where micelles aggregate to first form long tubules that become unstable, and break up into vesicles. A simple theory, based on energy principles, is presented to explain the tubule-vesicle transition. Observation of this new tubular intermediate state has been facilitated by the development of an integrated microfluidic chip/cryogenic transmission electron microscopy (cryo-TEM) unit. Although this transition has been observed in a specific amphiphilic system where micellar solutions of cetyltrimethylammonium bromide (CTAB) and dodecylbenzene sulfonic acid (HDBS) are mixed to form vesicles, this new tool can be applied broadly to study transient structures in nanoscale systems under the very controlled conditions provided by microfluidics.

  20. Microfluidic Lab-on-a-Chip Platforms: Requirements, Characteristics and Applications

    NASA Astrophysics Data System (ADS)

    Mark, D.; Haeberle, S.; Roth, G.; Von Stetten, F.; Zengerle, R.

    This review summarizes recent developments in microfluidic platform approaches. In contrast to isolated application-specific solutions, a microfluidic platform provides a set of fluidic unit operations, which are designed for easy combination within a well-defined fabrication technology. This allows the implementation of different application-specific (bio-) chemical processes, automated by microfluidic process integration [1]. A brief introduction into technical advances, major market segments and promising applications is followed by a detailed characterization of different microfluidic platforms, comprising a short definition, the functional principle, microfluidic unit operations, application examples as well as strengths and limitations. The microfluidic platforms in focus are lateral flow tests, linear actuated devices, pressure driven laminar flow, microfluidic large scale integration, segmented flow microfluidics, centrifugal microfluidics, electro-kinetics, electrowetting, surface acoustic waves, and systems for massively parallel analysis. The review concludes with the attempt to provide a selection scheme for microfluidic platforms which is based on their characteristics according to key requirements of different applications and market segments. Applied selection criteria comprise portability, costs of instrument and disposable, sample throughput, number of parameters per sample, reagent consumption, precision, diversity of microfluidic unit operations and the flexibility in programming different liquid handling protocols.

  1. Separation of carboxylic acids in human serum by isotachophoresis using a commercial field-deployable analytical platform combined with in-house glass microfluidic chips.

    PubMed

    Smejkal, Petr; Breadmore, Michael C; Guijt, Rosanne M; Grym, Jakub; Foret, František; Bek, Fritz; Macka, Mirek

    2012-11-28

    Portable and field deployable analytical instruments are attractive in many fields including medical diagnostics, where point of care and on-site diagnostics systems capable of providing rapid quantitative results have the potential to vastly improve the productivity and the quality of medical care. Isotachophoresis (ITP) is a well known electrophoretic separation technique previously demonstrated as suitable for miniaturization in microfluidic chip format (chip-ITP). In this work, a purpose-designed ITP chip compatible with a commercial end-used targeted microfluidic system was used to study different injection protocols and to evaluate the effect of the length of the separation channel on the analytical performance. The in-house ITP chips were made from Corning glass and compared to the commercial DNA chip for the ITP separation of anions from the hydrodynamic injection of human serum. Using the in-house ITP chip the isotachophoretic step of lactate from human serum was approximately two times longer. The results of this research suggested that microfluidic ITP with indirect fluorescence detection is a viable technique for separation of organic acids in human serum samples, especially when a chip with suitable design is used.

  2. Study of endothelial cell apoptosis using fluorescence resonance energy transfer (FRET) biosensor cell line with hemodynamic microfluidic chip system.

    PubMed

    Yu, J Q; Liu, X F; Chin, L K; Liu, A Q; Luo, K Q

    2013-07-21

    To better understand how hyperglycemia induces endothelial cell dysfunction under the diabetic conditions, a hemodynamic microfluidic chip system was developed. The system combines a caspase-3-based fluorescence resonance energy transfer (FRET) biosensor cell line which can detect endothelial cell apoptosis in real-time, post-treatment effect and with a limited cell sample, by using a microfluidic chip which can mimic the physiological pulsatile flow profile in the blood vessel. The caspase-3-based FRET biosensor endothelial cell line (HUVEC-C3) can produce a FRET-based sensor protein capable of probing caspase-3 activation. When the endothelial cells undergo apoptosis, the color of the sensor cells changes from green to blue, thus sensing apoptosis. A double-labeling fluorescent technique (yo pro-1 and propidium iodide) was used to validate the findings revealed by the FRET-based caspase sensor. The results show high rates of apoptosis and necrosis of endothelial cells when high glucose concentration was applied in our hemodynamic microfluidic chip combined with an exhaustive pulsatile flow profile. The two apoptosis detection techniques (fluorescent method and FRET biosensor) are comparable; but FRET biosensor offers more advantages such as real-time observation and a convenient operating process to generate more accurate and reliable data. Furthermore, the activation of the FRET biosensor also confirms the endothelial cell apoptosis induced by the abnormal pulsatile shear stress and high glucose concentration is through caspase-3 pathway. A 12% apoptotic rate (nearly a 4-fold increase compared to the static condition) was observed when the endothelial cells were exposed to a high glucose concentration of 20 mM under 2 h exhaustive pulsatile shear stress of 30 dyne cm(-2) and followed with another 10 h normal pulsatile shear stress of 15 dyne cm(-2). Therefore, the most important finding of this study is to develop a novel endothelial cell apoptosis detection

  3. Flow-orthogonal bead oscillation in a microfluidic chip with a magnetic anisotropic flux-guide array.

    PubMed

    van Pelt, Stijn; Derks, Roy; Matteucci, Marco; Hansen, Mikkel Fougt; Dietzel, Andreas

    2011-04-01

    A new concept for the manipulation of superparamagnetic beads inside a microfluidic chip is presented in this paper. The concept allows for bead actuation orthogonal to the flow direction inside a microchannel. Basic manipulation functionalities were studied by means of finite element simulations and results were oval-shaped steady state oscillations with bead velocities up to 500 μm/s. The width of the trajectory could be controlled by prescribing external field rotation. Successful verification experiments were performed on a prototype chip fabricated with excimer laser ablation in polycarbonate and electroforming of nickel flux-guides. Bead velocities up to 450 μm/s were measured in a 75 μm wide channel. By prescribing the currents in the external quadrupole magnet, the shape of the bead trajectory could be controlled.

  4. Automation of daphtoxkit-F biotest using a microfluidic lab-on-a-chip technology

    NASA Astrophysics Data System (ADS)

    Huang, Yushi; Nugegoda, Dayanthi; Wlodkowic, Donald

    2015-12-01

    An increased rigor in water quality monitoring is not only a legal requirement, but is also critical to ensure timely chemical hazard emergency responses and protection of human and animal health. Bioindication is a method that applies very sensitive living organisms to detect environmental changes using their natural responses. Although bioindicators do not deliver information on an exact type or intensity of toxicants present in water samples, they do provide an overall snapshot and early-warning information about presence of harmful and dangerous parameters. Despite the advantages of biotests performed on sentinel organisms, their wider application is limited by the nonexistence of high-throughput laboratory automation systems. As a result majority of biotests used in ecotoxicology require time-consuming and laborious manual procedures. In this work, we present development of a miniaturized Lab-on-a-Chip (LOC) platform for automation and enhancement of acute ecotoxicity test based on immobilization of a freshwater crustacean Daphnia magna (Daphtoxkit-FTM). Daphnids' immobilization in response to sudden changes in environment parameters is fast, unambiguous, and easy to record optically. We also for the first time demonstrate that LOC system enables studies of sub-lethal ecotoxic effects using behavioral responses of Daphnia magna as sentinels of water pollution. The system working principle incorporated a high definition (HD) time-resolved video data analysis to dynamically assess impact of the reference toxicant on swimming behavior of D. magna. Our system design combined: (i) microfluidic device for caging of Daphnia sp.; (ii) mechatronic interface for fluidic actuation; (iii) video data acquisition; and (iv) algorithms for animal movement tracking and analysis.

  5. A large format membrane-based x-ray mask for microfluidic chip fabrication

    NASA Astrophysics Data System (ADS)

    Wang, Lin; Zhang, Min; Desta, Yohannes; Melzak, J.; Wu, C. H.; Peng, Zhengchun

    2006-02-01

    X-ray lithography is a very good option for the fabrication of micro-devices especially when high aspect ratio patterns are required. Membrane-based x-ray masks are commonly used for high-resolution x-ray lithography. A thin layer of silicon nitride (Si3N4) or silicon carbide (SiC) film (1-2 µm) is normally used as the membrane material for x-ray mask fabrication (Wells G M, Reilly M, Nachman R, Cerrina F, El-Khakani M A and Chaker M 1993 Mater. Res. Soc. Conf. Proc. 306 81-9 Shoki T, Nagasawa H, Kosuga H, Yamaguchi Y, Annaka N, Amemiya I and Nagarekawa O 1993 SPIE Proc. 1924 450-6). The freestanding membrane window of an x-ray mask, which defines the exposing area of the x-ray mask, can be obtained by etching a pre-defined area on a silicon wafer from the backside (Wang L, Desta Y, Fettig R K, Goettert J, Hein H, Jakobs P and Chulz J 2004 J. Micromech. Microeng. 14 722-6). Usually, the window size of an x-ray mask is around 20 × 20 mm because of the low tensile stress of the membrane (10-100 MPa), and the larger window dimension of an x-ray mask may cause the deformation of membranes and lower the mask quality. However, x-ray masks with larger windows are preferred for micro-device fabrication in order to increase the productivity. We analyzed the factors which influence the flatness of large format x-ray masks and fabricated x-ray masks with a window size of 55 × 55 mm and 46 × 65 mm on 1 µm thick membranes by increasing the tensile stress of the membranes (>300 MPa) and optimizing the stress of the absorber layer. The large format x-ray mask was successfully applied for the fabrication of microfluidic chips.

  6. Microfluidic Chip-Based Detection and Intraspecies Strain Discrimination of Salmonella Serovars Derived from Whole Blood of Septic Mice

    PubMed Central

    Patterson, Adriana S.; Heithoff, Douglas M.; Ferguson, Brian S.; Soh, H. Tom; Mahan, Michael J.

    2013-01-01

    Salmonella is a zoonotic pathogen that poses a considerable public health and economic burden in the United States and worldwide. Resultant human diseases range from enterocolitis to bacteremia to sepsis and are acutely dependent on the particular serovar of Salmonella enterica subsp. enterica, which comprises over 99% of human-pathogenic S. enterica isolates. Point-of-care methods for detection and strain discrimination of Salmonella serovars would thus have considerable benefit to medical, veterinary, and field applications that safeguard public health and reduce industry-associated losses. Here we describe a single, disposable microfluidic chip that supports isothermal amplification and sequence-specific detection and discrimination of Salmonella serovars derived from whole blood of septic mice. The integrated microfluidic electrochemical DNA (IMED) chip consists of an amplification chamber that supports loop-mediated isothermal amplification (LAMP), a rapid, single-temperature amplification method as an alternative to PCR that offers advantages in terms of sensitivity, reaction speed, and amplicon yield. The amplification chamber is connected via a microchannel to a detection chamber containing a reagentless, multiplexed (here biplex) sensing array for sequence-specific electrochemical DNA (E-DNA) detection of the LAMP products. Validation of the IMED device was assessed by the detection and discrimination of S. enterica subsp. enterica serovars Typhimurium and Choleraesuis, the causative agents of enterocolitis and sepsis in humans, respectively. IMED chips conferred rapid (under 2 h) detection and discrimination of these strains at clinically relevant levels (<1,000 CFU/ml) from whole, unprocessed blood collected from septic animals. The IMED-based chip assay shows considerable promise as a rapid, inexpensive, and portable point-of-care diagnostic platform for the detection and strain-specific discrimination of microbial pathogens. PMID:23354710

  7. Hybrid microfluidic systems: combining a polymer microfluidic toolbox with biosensors

    NASA Astrophysics Data System (ADS)

    Gärtner, Claudia; Kirsch, Stefanie; Anton, Birgit; Becker, Holger

    2007-01-01

    In this paper we present polymer based microfluidic chips which contain functional elements (electrodes, biosensors) made out of a different material (metals, silicon, organic semiconductors). These hybrid microfluidic devices allow the integration of additional functionality other than the simple manipulation of liquids in the chip and have been developed as a reaction to the increasing requirement for functional integration in microfluidics.

  8. A 3D microfluidic chip for electrochemical detection of hydrolysed nucleic bases by a modified glassy carbon electrode.

    PubMed

    Vlachova, Jana; Tmejova, Katerina; Kopel, Pavel; Korabik, Maria; Zitka, Jan; Hynek, David; Kynicky, Jindrich; Adam, Vojtech; Kizek, Rene

    2015-01-22

    Modification of carbon materials, especially graphene-based materials, has wide applications in electrochemical detection such as electrochemical lab-on-chip devices. A glassy carbon electrode (GCE) modified with chemically alternated graphene oxide was used as a working electrode (glassy carbon modified by graphene oxide with sulphur containing compounds and Nafion) for detection of nucleobases in hydrolysed samples (HCl pH = 2.9, 100 °C, 1 h, neutralization by NaOH). It was found out that modification, especially with trithiocyanuric acid, increased the sensitivity of detection in comparison with pure GCE. All processes were finally implemented in a microfluidic chip formed with a 3D printer by fused deposition modelling technology. As a material for chip fabrication, acrylonitrile butadiene styrene was chosen because of its mechanical and chemical stability. The chip contained the one chamber for the hydrolysis of the nucleic acid and another for the electrochemical detection by the modified GCE. This chamber was fabricated to allow for replacement of the GCE.

  9. A 3D Microfluidic Chip for Electrochemical Detection of Hydrolysed Nucleic Bases by a Modified Glassy Carbon Electrode

    PubMed Central

    Vlachova, Jana; Tmejova, Katerina; Kopel, Pavel; Korabik, Maria; Zitka, Jan; Hynek, David; Kynicky, Jindrich; Adam, Vojtech; Kizek, Rene

    2015-01-01

    Modification of carbon materials, especially graphene-based materials, has wide applications in electrochemical detection such as electrochemical lab-on-chip devices. A glassy carbon electrode (GCE) modified with chemically alternated graphene oxide was used as a working electrode (glassy carbon modified by graphene oxide with sulphur containing compounds and Nafion) for detection of nucleobases in hydrolysed samples (HCl pH = 2.9, 100 °C, 1 h, neutralization by NaOH). It was found out that modification, especially with trithiocyanuric acid, increased the sensitivity of detection in comparison with pure GCE. All processes were finally implemented in a microfluidic chip formed with a 3D printer by fused deposition modelling technology. As a material for chip fabrication, acrylonitrile butadiene styrene was chosen because of its mechanical and chemical stability. The chip contained the one chamber for the hydrolysis of the nucleic acid and another for the electrochemical detection by the modified GCE. This chamber was fabricated to allow for replacement of the GCE. PMID:25621613

  10. Creating Sub-50 Nm Nanofluidic Junctions in PDMS Microfluidic Chip via Self-Assembly Process of Colloidal Particles.

    PubMed

    Wei, Xi; Syed, Abeer; Mao, Pan; Han, Jongyoon; Song, Yong-Ak

    2016-03-13

    Polydimethylsiloxane (PDMS) is the prevailing building material to make microfluidic devices due to its ease of molding and bonding as well as its transparency. Due to the softness of the PDMS material, however, it is challenging to use PDMS for building nanochannels. The channels tend to collapse easily during plasma bonding. In this paper, we present an evaporation-driven self-assembly method of silica colloidal nanoparticles to create nanofluidic junctions with sub-50 nm pores between two microchannels. The pore size as well as the surface charge of the nanofluidic junction is tunable simply by changing the colloidal silica bead size and surface functionalization outside of the assembled microfluidic device in a vial before the self-assembly process. Using the self-assembly of nanoparticles with a bead size of 300 nm, 500 nm, and 900 nm, it was possible to fabricate a porous membrane with a pore size of ~45 nm, ~75 nm and ~135 nm, respectively. Under electrical potential, this nanoporous membrane initiated ion concentration polarization (ICP) acting as a cation-selective membrane to concentrate DNA by ~1,700 times within 15 min. This non-lithographic nanofabrication process opens up a new opportunity to build a tunable nanofluidic junction for the study of nanoscale transport processes of ions and molecules inside a PDMS microfluidic chip.

  11. A compound magnetic field generating system for targeted killing of Staphylococcus aureus by magnetotactic bacteria in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Chen, Linjie; Chen, Changyou; Wang, Pingping; Chen, Chuanfang; Wu, Long-Fei; Song, Tao

    2017-04-01

    A compound magnetic field generating system was built to kill Staphylococcus aureus (S. aureus) by magnetotactic bacteria (MTB) in a microfluidic chip in this paper. The system was consisted of coil pairs, a switch circuit, a control program and controllable electrical sources. It could produce a guiding magnetic field (gMF) of ±1 mT along arbitrary direction in the horizontal plane, a rotating magnetic field (rMF) and a swing magnetic field (sMF, 2 Hz, 10 mT) by controlling the currents. The gMF was used to guide MTB swimming to the S. aureus pool in the microfluidic chip, and then the rMF enhanced the mixture of S. aureus and MTB cells, therefore beneficial to the attachments of them. Finally, the sMF was used to induce the death of S. aureus via MTB. The results showed that MTB could be navigated by the gMF and that 47.1% of S. aureus were killed when exposed to the sMF. It provides a new solution for the targeted treatment of infected diseases and even cancers.

  12. High efficiency integration of three-dimensional functional microdevices inside a microfluidic chip by using femtosecond laser multifoci parallel microfabrication

    NASA Astrophysics Data System (ADS)

    Xu, Bing; Du, Wen-Qiang; Li, Jia-Wen; Hu, Yan-Lei; Yang, Liang; Zhang, Chen-Chu; Li, Guo-Qiang; Lao, Zhao-Xin; Ni, Jin-Cheng; Chu, Jia-Ru; Wu, Dong; Liu, Su-Ling; Sugioka, Koji

    2016-01-01

    High efficiency fabrication and integration of three-dimension (3D) functional devices in Lab-on-a-chip systems are crucial for microfluidic applications. Here, a spatial light modulator (SLM)-based multifoci parallel femtosecond laser scanning technology was proposed to integrate microstructures inside a given ‘Y’ shape microchannel. The key novelty of our approach lies on rapidly integrating 3D microdevices inside a microchip for the first time, which significantly reduces the fabrication time. The high quality integration of various 2D-3D microstructures was ensured by quantitatively optimizing the experimental conditions including prebaking time, laser power and developing time. To verify the designable and versatile capability of this method for integrating functional 3D microdevices in microchannel, a series of microfilters with adjustable pore sizes from 12.2 μm to 6.7 μm were fabricated to demonstrate selective filtering of the polystyrene (PS) particles and cancer cells with different sizes. The filter can be cleaned by reversing the flow and reused for many times. This technology will advance the fabrication technique of 3D integrated microfluidic and optofluidic chips.

  13. A coral-on-a-chip microfluidic platform enabling live-imaging microscopy of reef-building corals

    PubMed Central

    Shapiro, Orr H.; Kramarsky-Winter, Esti; Gavish, Assaf R.; Stocker, Roman; Vardi, Assaf

    2016-01-01

    Coral reefs, and the unique ecosystems they support, are facing severe threats by human activities and climate change. Our understanding of these threats is hampered by the lack of robust approaches for studying the micro-scale interactions between corals and their environment. Here we present an experimental platform, coral-on-a-chip, combining micropropagation and microfluidics to allow direct microscopic study of live coral polyps. The small and transparent coral micropropagates are ideally suited for live-imaging microscopy, while the microfluidic platform facilitates long-term visualization under controlled environmental conditions. We demonstrate the usefulness of this approach by imaging coral micropropagates at previously unattainable spatio-temporal resolutions, providing new insights into several micro-scale processes including coral calcification, coral–pathogen interaction and the loss of algal symbionts (coral bleaching). Coral-on-a-chip thus provides a powerful method for studying coral physiology in vivo at the micro-scale, opening new vistas in coral biology. PMID:26940983

  14. High-throughput and clogging-free microfluidic filtration platform for on-chip cell separation from undiluted whole blood

    PubMed Central

    Cheng, Yinuo; Ye, Xiongying; Ma, Zengshuai; Xie, Shuai; Wang, Wenhui

    2016-01-01

    Rapid separation of white blood cells from whole blood sample is often required for their subsequent analyses of functions and phenotypes, and many advances have been made in this field. However, most current microfiltration-based cell separation microfluidic chips still suffer from low-throughput and membrane clogging. This paper reports on a high-throughput and clogging-free microfluidic filtration platform, which features with an integrated bidirectional micropump and commercially available polycarbonate microporous membranes. The integrated bidirectional micropump enables the fluid to flush micropores back and forth, effectively avoiding membrane clogging. The microporous membrane allows red blood cells passing through high-density pores in a cross-flow mixed with dead-end filtration mode. All the separation processes, including blood and buffer loading, separation, and sample collection, are automatically controlled for easy operation and high throughput. Both microbead mixture and undiluted whole blood sample are separated by the platform effectively. In particular, for white blood cell separation, the chip recovered 72.1% white blood cells with an over 232-fold enrichment ratio at a throughput as high as 37.5 μl/min. This high-throughput, clogging-free, and highly integrated platform holds great promise for point-of-care blood pretreatment, analysis, and diagnosis applications. PMID:26909124

  15. 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.

  16. High efficiency integration of three-dimensional functional microdevices inside a microfluidic chip by using femtosecond laser multifoci parallel microfabrication

    PubMed Central

    Xu, Bing; Du, Wen-Qiang; Li, Jia-Wen; Hu, Yan-Lei; Yang, Liang; Zhang, Chen-Chu; Li, Guo-Qiang; Lao, Zhao-Xin; Ni, Jin-Cheng; Chu, Jia-Ru; Wu, Dong; Liu, Su-Ling; Sugioka, Koji

    2016-01-01

    High efficiency fabrication and integration of three-dimension (3D) functional devices in Lab-on-a-chip systems are crucial for microfluidic applications. Here, a spatial light modulator (SLM)-based multifoci parallel femtosecond laser scanning technology was proposed to integrate microstructures inside a given ‘Y’ shape microchannel. The key novelty of our approach lies on rapidly integrating 3D microdevices inside a microchip for the first time, which significantly reduces the fabrication time. The high quality integration of various 2D-3D microstructures was ensured by quantitatively optimizing the experimental conditions including prebaking time, laser power and developing time. To verify the designable and versatile capability of this method for integrating functional 3D microdevices in microchannel, a series of microfilters with adjustable pore sizes from 12.2 μm to 6.7 μm were fabricated to demonstrate selective filtering of the polystyrene (PS) particles and cancer cells with different sizes. The filter can be cleaned by reversing the flow and reused for many times. This technology will advance the fabrication technique of 3D integrated microfluidic and optofluidic chips. PMID:26818119

  17. High-throughput and clogging-free microfluidic filtration platform for on-chip cell separation from undiluted whole blood.

    PubMed

    Cheng, Yinuo; Ye, Xiongying; Ma, Zengshuai; Xie, Shuai; Wang, Wenhui

    2016-01-01

    Rapid separation of white blood cells from whole blood sample is often required for their subsequent analyses of functions and phenotypes, and many advances have been made in this field. However, most current microfiltration-based cell separation microfluidic chips still suffer from low-throughput and membrane clogging. This paper reports on a high-throughput and clogging-free microfluidic filtration platform, which features with an integrated bidirectional micropump and commercially available polycarbonate microporous membranes. The integrated bidirectional micropump enables the fluid to flush micropores back and forth, effectively avoiding membrane clogging. The microporous membrane allows red blood cells passing through high-density pores in a cross-flow mixed with dead-end filtration mode. All the separation processes, including blood and buffer loading, separation, and sample collection, are automatically controlled for easy operation and high throughput. Both microbead mixture and undiluted whole blood sample are separated by the platform effectively. In particular, for white blood cell separation, the chip recovered 72.1% white blood cells with an over 232-fold enrichment ratio at a throughput as high as 37.5 μl/min. This high-throughput, clogging-free, and highly integrated platform holds great promise for point-of-care blood pretreatment, analysis, and diagnosis applications.

  18. Application of a multi-channel microfluidic chip on the simultaneous detection of DNAs by using microbead-quantum dots.

    PubMed

    Le, Ngoc Tam; Kim, Jong Sung

    2014-12-01

    Several researches have shown that cancer is caused by genetic mutations especially in genes involved in cell growth and regulation. Ras family members are frequently found in their mutated, oncogenic forms in human tumors. Mutant RAS proteins are constitutively active, owing to reduce intrinsic GTPase activity and insensitivity to GTPase-activating protein (GAPs). In total, activating mutations in the RAS genes occur in approximately 20% of all human cancers, mainly in codon 12, 13 or 61. Activating mutations in the NRAS gene not only result in the reduction of intrinsic GTPase activity but also in the induction of resistance against molecules inducing such activity. In this paper, we reported a rapid, simple and portable method for detecting the mutant types of NRAS genes codon 12 and 61 simultaneously by using bead-quantum dots (QDs) based multi-channel microfluidic chip. Probe DNAs are conjugated to bead-QDs and packed in the pillars of channels in the microfluidic chip. After injection of target DNAs and intercalating dyes, the fluorescence quenching of QDs by intercalating dye was observed due to FRET phenomena. The platform can be effortlessly applied in other biological and clinical areas.

  19. Chemiluminescence determination of moxifloxacin in pharmaceutical and biological samples based on its enhancing effect of the luminol-ferricyanide system using a microfluidic chip.

    PubMed

    Suh, Yeoun Suk; Kamruzzaman, Mohammad; Alam, Al-Mahmnur; Lee, Sang Hak; Kim, Young Ho; Kim, Gyu-Man; Dang, Trung Dung

    2014-05-01

    A sensitive determination of a synthetic fluoroquinolone antibacterial agent, moxifloxacin (MOX), by an enhanced chemiluminescence (CL) method using a microfluidic chip is described. The microfluidic chip was fabricated by a soft-lithographic procedure using polydimethyl siloxane (PDMS). The fabricated PDMS microfluidic chip had three-inlet microchannels for introducing the sample, chemiluminescent reagent and oxidant, and a 500 µm wide, 250 µm deep and 82 mm long microchannel. An enhanced CL system, luminol-ferricyanide, was adopted to analyze the MOX concentration in a sample solution. CL light was emitted continuously after mixing luminol and ferricyanide in the presence of MOX on the PDMS microfluidic chip. The amount of MOX in the luminol-ferricyanide system influenced the intensity of the CL light. The linear range of MOX concentration was 0.14-55.0 ng/mL with a correlation coefficient of 0.9992. The limit of detection (LOD) and limit of quantification (LOQ) were 0.06 and 0.2 ng/mL respectively. The presented method afforded good reproducibility, with a relative standard deviation (RSD) of 1.05% for 10 ng/mL of MOX, and has been successfully applied for the determination of MOX in pharmaceutical and biological samples.

  20. Co-Culture of Tumor Spheroids and Fibroblasts in a Collagen Matrix-Incorporated Microfluidic Chip Mimics Reciprocal Activation in Solid Tumor Microenvironment

    PubMed Central

    Jeong, Su-Yeong; Lee, Ji-Hyun; Shin, Yoojin; Chung, Seok; Kuh, Hyo-Jeong

    2016-01-01

    Multicellular 3D culture and interaction with stromal components are considered essential elements in establishing a ‘more clinically relevant’ tumor model. Matrix-embedded 3D cultures using a microfluidic chip platform can recapitulate the microscale interaction within tumor microenvironments. As a major component of tumor microenvironment, cancer-associated fibroblasts (CAFs) play a role in cancer progression and drug resistance. Here, we present a microfluidic chip-based tumor tissue culture model that integrates 3D tumor spheroids (TSs) with CAF in proximity within a hydrogel scaffold. HT-29 human colorectal carcinoma cells grew into 3D TSs and the growth was stimulated when co-cultured with fibroblasts as shown by 1.5-folds increase of % changes in diameter over 5 days. TS cultured for 6 days showed a reduced expression of Ki-67 along with increased expression of fibronectin when co-cultured with fibroblasts compared to mono-cultured TSs. Fibroblasts were activated under co-culture conditions, as demonstrated by increases in α-SMA expression and migratory activity. When exposed to paclitaxel, a survival advantage was observed in TSs co-cultured with activated fibroblasts. Overall, we demonstrated the reciprocal interaction between TSs and fibroblasts in our 7-channel microfluidic chip. The co-culture of 3D TS-CAF in a collagen matrix-incorporated microfluidic chip may be useful to study the tumor microenvironment and for evaluation of drug screening and evaluation. PMID:27391808

  1. In-Depth Characterization of N-Linked Oligosaccharides Using Fluoride-Mediated Negative Ion Microfluidic Chip LC-MS

    PubMed Central

    Ni, Wenqin; Bones, Jonathan; Karger, Barry L.

    2013-01-01

    Characterization of N-glycans by liquid chromatography-positive electrospray ionization (ESI) tandem mass spectrometry (LC-MS/MS) using a microfluidic chip packed with porous graphitized carbon (PGC) represents a rapidly developing area in oligosaccharide analysis. Positive ion ESI-MS generates B/Y-type glycosidic fragment ions under collisional induced dissociation (CID). Although these ions facilitate glycan sequencing, they provide little information on linkage and positional isomers. Isomer identification in these cases is by retention on the PGC stationary phase where the specific structural isomers can, in principle, be separated. In this paper, we broaden the applicability of the PGC microfluidic chip/MS platform by implementing fluoride-mediated negative ESI-MS. Ammonium fluoride, added to the mobile phase, aids in the formation of pseudomolecular oligosaccharide anions due to the ability of fluoride to abstract a proton from the glycan structure. The negative charge results in the generation of C-type glycosidic fragments, highly informative A-type cross ring fragment ions and additional gas phase ion reaction products (e.g., D- and E-type ions), which, when combined, lead to in-depth oligosaccharide characterization, including linkage and positional isomers. Due to the separation of anomers by the PGC phase, comparison of oligosaccharides with an intact reducing terminus to their corresponding alditols was performed, revealing a more sensitive MS and, especially, MS/MS response from the glycans with a free reducing end. Fluoride also ensured recovery of charged oligosaccharides from the PGC stationary phase. Application to the characterization of N-glycans released from polyclonal human and murine serum IgG is presented to demonstrate the effectiveness of the chip/negative ESI approach. PMID:23398125

  2. Coupling paper-based microfluidics and lab on a chip technologies for confirmatory analysis of trinitro aromatic explosives.

    PubMed

    Pesenti, Alessandra; Taudte, Regina Verena; McCord, Bruce; Doble, Philip; Roux, Claude; Blanes, Lucas

    2014-05-20

    A new microfluidic paper-based analytical device (μPAD) in conjunction with confirmation by a lab on chip analysis was developed for detection of three trinitro aromatic explosives. Potassium hydroxide was deposited on the μPADs (0.5 μL, 1.5 M), creating a color change reaction when explosives are present, with detection limits of approximately 7.5 ± 1.0 ng for TNB, 12.5 ± 2.0 ng for TNT and 15.0 ± 2.0 ng for tetryl. For confirmatory analysis, positive μPADs were sampled using a 5 mm hole-punch, followed by extraction of explosives from the punched chad in 30 s using 20 μL borate/SDS buffer. The extractions had efficiencies of 96.5 ± 1.7%. The extracted explosives were then analyzed with the Agilent 2100 Bioanalyzer lab on a chip device with minimum detectable amounts of 3.8 ± 0.1 ng for TNB, 7.0 ± 0.9 ng for TNT, and 4.7 ± 0.2 ng for tetryl. A simulated in-field scenario demonstrated the feasibility of coupling the μPAD technique with the lab on a chip device to detect and identify 1 μg of explosives distributed on a surface of 100 cm(2).

  3. Rapid on-chip recalcification and drug dosing of citrated whole blood using microfluidic buffer sheath flow.

    PubMed

    Muthard, Ryan W; Diamond, Scott L

    2014-01-01

    Millions of clotting tests each year require recalcification of blood treated with sodium citrate, a calcium chelator that prevents prothrombinase assembly. We validated a converging trifurcated microfluidic device to measure platelet and fibrin accumulation following on-chip recalcification of citrated whole blood. Recalcification was accomplished by sheathing the blood with Ca2+ buffer. Fluorescein rapidly diffused across the buffer-blood interface (achieving 62.5% of maximum centerline concentration within ~4 cm of flow), while albumin remained relatively unchanged in blood due to its lower diffusivity (<20% decrease). Since Ca2+ diffuses faster than fluorescein, full recalcification of whole blood was achieved within ~1 cm of flow prior to encountering a collagen/tissue surface. Platelet and fibrin were reduced by 87.3% and 99.0%, respectively, when the sheath buffer was Ca2+-free. A 30-min preincubation of citrated whole blood prior to on-chip recalcification increased platelet (159%) and fibrin (86.6%) deposition, compared to 5-min preincubation, likely due to factor XIIa generation in citrated blood. The P2Y1 inhibitor, MRS-2179, was delivered by diffusion into flowing blood and inhibited platelet deposition on collagen with a calculated IC50 of 0.155 μM. On-chip recalcification and drug dosing of citrated blood allows for assays of platelet function in a whole blood milieu under flow.

  4. Brain slice on a chip: opportunities and challenges of applying microfluidic technology to intact tissues.

    PubMed

    Huang, Yu; Williams, Justin C; Johnson, Stephen M

    2012-06-21

    Isolated brain tissue, especially brain slices, are valuable experimental tools for studying neuronal function at the network, cellular, synaptic, and single channel levels. Neuroscientists have refined the methods for preserving brain slice viability and function and converged on principles that strongly resemble the approach taken by engineers in developing microfluidic devices. With respect to brain slices, microfluidic technology may 1) overcome the traditional limitations of conventional interface and submerged slice chambers and improve oxygen/nutrient penetration into slices, 2) provide better spatiotemporal control over solution flow/drug delivery to specific slice regions, and 3) permit successful integration with modern optical and electrophysiological techniques. In this review, we highlight the unique advantages of microfluidic devices for in vitro brain slice research, describe recent advances in the integration of microfluidic devices with optical and electrophysiological instrumentation, and discuss clinical applications of microfluidic technology as applied to brain slices and other non-neuronal tissues. We hope that this review will serve as an interdisciplinary guide for both neuroscientists studying brain tissue in vitro and engineers as they further develop microfluidic chamber technology for neuroscience research.

  5. Capillary-driven microfluidic paper-based analytical devices for lab on a chip screening of explosive residues in soil.

    PubMed

    Ueland, Maiken; Blanes, Lucas; Taudte, Regina V; Stuart, Barbara H; Cole, Nerida; Willis, Peter; Roux, Claude; Doble, Philip

    2016-03-04

    A novel microfluidic paper-based analytical device (μPAD) was designed to filter, extract, and pre-concentrate explosives from soil for direct analysis by a lab on a chip (LOC) device. The explosives were extracted via immersion of wax-printed μPADs directly into methanol soil suspensions for 10min, whereby dissolved explosives travelled upwards into the μPAD circular sampling reservoir. A chad was punched from the sampling reservoir and inserted into a LOC well containing the separation buffer for direct analysis, avoiding any further extraction step. Eight target explosives were separated and identified by fluorescence quenching. The minimum detectable amounts for all eight explosives were between 1.4 and 5.6ng with recoveries ranging from 53-82% from the paper chad, and 12-40% from soil. This method provides a robust and simple extraction method for rapid identification of explosives in complex soil samples.

  6. A portable instrument for continuous glucose monitoring by the integration of microfluidic chip and micro-glucose sensor

    NASA Astrophysics Data System (ADS)

    Li, Dachao; Ji, Yongjie; Liang, Wenshuai; Zhang, Xiaoli; Yu, Haixia; Xu, Kexin

    2013-03-01

    Interstitial fluid (ISF) can be transdermally extracted using low-frequency ultrasound and continuous vacuum pressure on skin surface. But the tiny volume of transdermally extracted ISF makes the transdermal extraction, collection, transport, volumetric detection and glucose concentration measurement of the ISF very difficult. Based on a microfluidic chip for transdermally extraction of interstitial fluid and a micro glucose sensor for glucose concentration measurement, a continuous glucose monitoring instrumentby ISF transdermal extraction with minimally invasive way is developed. In the paper, various parts of the device and their interface circuits are designed; the hardware and software of the instrument are built; the simulating experiments of transdermal ISF extraction, collection and volume measurement with full-thickness pig skin are performed using this integrated system; and the functionalities of this device is verified for future clinical application.

  7. Design of a Microfluidic Chip for Magnetic-Activated Sorting of One-Bead-One-Compound Libraries.

    PubMed

    Cho, Choi-Fong; Lee, Kyungheon; Speranza, Maria-Carmela; Bononi, Fernanda C; Viapiano, Mariano S; Luyt, Leonard G; Weissleder, Ralph; Chiocca, E Antonio; Lee, Hakho; Lawler, Sean E

    2016-06-13

    Molecular targeting using ligands specific to disease markers has shown great promise for early detection and directed therapy. Bead-based combinatorial libraries have served as powerful tools for the discovery of novel targeting agents. Screening platforms employing magnetic capture have been used to achieve rapid and efficient identification of high-affinity ligands from one-bead-one-compound (OBOC) libraries. Traditional manual methodologies to isolate magnetized "hit" beads are tedious and lack accuracy, and existing instruments to expedite bead sorting tend to be costly and complex. Here, we describe the design and construction of a simple and inexpensive microfluidic magnetic sorting device using standard photolithography and soft lithography approaches to facilitate high-throughput isolation of magnetized positive hit beads from combinatorial libraries. We have demonstrated that the device is able to sort magnetized beads with superior accuracy compared to conventional manual sorting approaches. This chip offers a very convenient yet inexpensive alternative for screening OBOC libraries.

  8. Fabrication of universal serial bus flash disk type microfluidic chip electrophoresis and application for protein analysis under ultra low voltage.

    PubMed

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Liu, Huwei; Yuan, Hua

    2016-03-01

    A simple and effective universal serial bus (USB) flash disk type microfluidic chip electrophoresis (MCE) was developed by using poly(dimethylsiloxane) based soft lithography and dry film based printed circuit board etching techniques in this paper. The MCE had a microchannel diameter of 375 μm and an effective length of 25 mm. Equipped with a conventional online electrochemical detector, the device enabled effectively separation of bovine serum albumin, lysozyme, and cytochrome c in 80 s under the ultra low voltage from a computer USB interface. Compared with traditional capillary electrophoresis, the USB flash disk type MCE is not only portable and inexpensive but also fast with high separation efficiency.

  9. Fabrication of universal serial bus flash disk type microfluidic chip electrophoresis and application for protein analysis under ultra low voltage

    PubMed Central

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Liu, Huwei

    2016-01-01

    A simple and effective universal serial bus (USB) flash disk type microfluidic chip electrophoresis (MCE) was developed by using poly(dimethylsiloxane) based soft lithography and dry film based printed circuit board etching techniques in this paper. The MCE had a microchannel diameter of 375 μm and an effective length of 25 mm. Equipped with a conventional online electrochemical detector, the device enabled effectively separation of bovine serum albumin, lysozyme, and cytochrome c in 80 s under the ultra low voltage from a computer USB interface. Compared with traditional capillary electrophoresis, the USB flash disk type MCE is not only portable and inexpensive but also fast with high separation efficiency. PMID:27042249

  10. Capacitance variation induced by microfluidic two-phase flow across insulated interdigital electrodes in lab-on-chip devices.

    PubMed

    Dong, Tao; Barbosa, Cátia

    2015-01-26

    Microfluidic two-phase flow detection has attracted plenty of interest in various areas of biology, medicine and chemistry. This work presents a capacitive sensor using insulated interdigital electrodes (IDEs) to detect the presence of droplets in a microchannel. This droplet sensor is composed of a glass substrate, patterned gold electrodes and an insulation layer. A polydimethylsiloxane (PDMS) cover bonded to the multilayered structure forms a microchannel. Capacitance variation induced by the droplet passage was thoroughly investigated with both simulation and experimental work. Olive oil and deionized water were employed as the working fluids in the experiments to demonstrate the droplet sensor. The results show a good sensitivity of the droplet with the appropriate measurement connection. This capacitive droplet sensor is promising to be integrated into a lab-on-chip device for in situ monitoring/counting of droplets or bubbles.

  11. Fast single run of vanilla fingerprint markers on microfluidic-electrochemistry chip for confirmation of common frauds.

    PubMed

    Avila, Mónica; Zougagh, Mohammed; Escarpa, Alberto; Ríos, Angel

    2009-10-01

    A new strategy based on the fast separation of the fingerprint markers of Vanilla planifolia extracts and vanilla-related samples on microfluidic-electrochemistry chip is proposed. This methodology allowed the detection of all required markers for confirmation of common frauds in this field. The elution order was strategically connected with sequential sample screening and analyte confirmation steps, where first ethyl vanillin was detected to distinguish natural from adultered samples; second, vanillin as prominent marker in V. planifolia, but frequently added in its synthetic form; and third, the final detection of the fingerprint markers (p-hydroxybenzaldehyde, vanillic acid, and p-hydroxybenzoic acid) of V. planifolia with confirmation purposes. The reliability of the proposed methodology was demonstrated in the confirmation the natural or non-natural origin of vanilla in samples using V. planifolia extracts and other selected food samples containing this flavor.

  12. Microfluidic chip based micro RNA detection through the combination of fluorescence and surface enhanced Raman scattering techniques.

    PubMed

    Wang, Zhile; Zong, Shenfei; Wang, Zhuyuan; Wu, Lei; Chen, Peng; Yun, Binfeng; Cui, Yiping

    2017-03-10

    We present a novel microfluidic chip based method for the detection of micro RNA (miRNA) via the combination of fluorescence and surface enhanced Raman scattering (SERS) spectroscopies. First, silver nanoparticles (Ag NPs) are immobilized onto a glass slide, forming a SERS enhancing substrate. Then a specificially designed molecular beacon (MB) is attached to the SERS substrate. The 3' end of the MB is decorated with a thiol group to facilitate the attachment of the MB, while the 5' end of the MB is labeled with an organic dye 6-FAM, which is used both as the fluorophore and SERS reporter. In the absence of target miRNA, the MB will form a hairpin structure, making 6-FAM close to the Ag NPs. Hence, the fluorescence of 6-FAM will be quenched and the Raman signal of 6-FAM will be enhanced. On the contrary, with target miRNA present, hybridization between the miRNA and MB will unfold the MB and increase the distance between 6-FAM and the Ag NPs. Thus the fluorescence of 6-FAM will recover and the SERS signal of 6-FAM will decrease. So the target miRNA will simultaneously introduce opposite changing trends in the intensities of the fluorescence and SERS signals. By combining the opposite changes in the two optical spectra, an improved sensitivity and linearity toward the target miRNA is achieved as compared with using solely fluorescence or SERS. Moreover, introducing the microfluidic chip can reduce the reaction time, reagent dosage and complexity of detection. With the improved sensitivity and simplicity, we anticipate that the presented method can have great potential in the investigation of miRNA related diseases.

  13. Microfluidic chip based micro RNA detection through the combination of fluorescence and surface enhanced Raman scattering techniques

    NASA Astrophysics Data System (ADS)

    Wang, Zhile; Zong, Shenfei; Wang, Zhuyuan; Wu, Lei; Chen, Peng; Yun, Binfeng; Cui, Yiping

    2017-03-01

    We present a novel microfluidic chip based method for the detection of micro RNA (miRNA) via the combination of fluorescence and surface enhanced Raman scattering (SERS) spectroscopies. First, silver nanoparticles (Ag NPs) are immobilized onto a glass slide, forming a SERS enhancing substrate. Then a specificially designed molecular beacon (MB) is attached to the SERS substrate. The 3‧ end of the MB is decorated with a thiol group to facilitate the attachment of the MB, while the 5‧ end of the MB is labeled with an organic dye 6-FAM, which is used both as the fluorophore and SERS reporter. In the absence of target miRNA, the MB will form a hairpin structure, making 6-FAM close to the Ag NPs. Hence, the fluorescence of 6-FAM will be quenched and the Raman signal of 6-FAM will be enhanced. On the contrary, with target miRNA present, hybridization between the miRNA and MB will unfold the MB and increase the distance between 6-FAM and the Ag NPs. Thus the fluorescence of 6-FAM will recover and the SERS signal of 6-FAM will decrease. So the target miRNA will simultaneously introduce opposite changing trends in the intensities of the fluorescence and SERS signals. By combining the opposite changes in the two optical spectra, an improved sensitivity and linearity toward the target miRNA is achieved as compared with using solely fluorescence or SERS. Moreover, introducing the microfluidic chip can reduce the reaction time, reagent dosage and complexity of detection. With the improved sensitivity and simplicity, we anticipate that the presented method can have great potential in the investigation of miRNA related diseases.

  14. Low-temperature bonded glass-membrane microfluidic device for in vitro organ-on-a-chip cell culture models

    NASA Astrophysics Data System (ADS)

    Pocock, Kyall J.; Gao, Xiaofang; Wang, Chenxi; Priest, Craig; Prestidge, Clive A.; Mawatari, Kazuma; Kitamori, Takehiko; Thierry, Benjamin

    2015-12-01

    The integration of microfluidics with living biological systems has paved the way to the exciting concept of "organson- a-chip", which aims at the development of advanced in vitro models that replicate the key features of human organs. Glass based devices have long been utilised in the field of microfluidics but the integration of alternative functional elements within multi-layered glass microdevices, such as polymeric membranes, remains a challenge. To this end, we have extended a previously reported approach for the low-temperature bonding of glass devices that enables the integration of a functional polycarbonate porous membrane. The process was initially developed and optimised on specialty low-temperature bonding equipment (μTAS2001, Bondtech, Japan) and subsequently adapted to more widely accessible hot embosser units (EVG520HE Hot Embosser, EVG, Austria). The key aspect of this method is the use of low temperatures compatible with polymeric membranes. Compared to borosilicate glass bonding (650 °C) and quartz/fused silica bonding (1050 °C) processes, this method maintains the integrity and functionality of the membrane (Tg 150 °C for polycarbonate). Leak tests performed showed no damage or loss of integrity of the membrane for up to 150 hours, indicating sufficient bond strength for long term cell culture. A feasibility study confirmed the growth of dense and functional monolayers of Caco-2 cells within 5 days.

  15. Microfluidic Chip-Based Online Screening Coupled to Mass Spectrometry: Identification of Inhibitors of Thrombin and Factor Xa.

    PubMed

    Iyer, Janaki Krishnamoorthy; Otvos, Reka A; Kool, Jeroen; Kini, R Manjunatha

    2016-02-01

    Thrombin and factor Xa (FXa) are critical enzymes of the blood coagulation cascade and are excellent targets of anticoagulant agents. Natural sources present an array of anticoagulants that can be developed as antithrombotic drugs. High-resolution, online screening techniques have been developed for the identification of drug leads from complex mixtures. In this study, we have developed and optimized a microfluidic online screening technique coupled to nano-liquid chromatography (LC) and in parallel with a mass spectrometer for the identification of thrombin and FXa inhibitors in mixtures. Inhibitors eluting from the nano-LC were split postcolumn in a 1:1 ratio; half was fed into a mass spectrometer (where its mass is detected), and the other half was fed into a microfluidic chip (which acts as a microreactor for the online assays). With our platform, thrombin and FXa inhibitors were detected in the assay in parallel with their mass identification. These methods are suitable for the identification of inhibitors from sample amounts as low as sub-microliter volumes.

  16. Stable, Free-space Optical Trapping and Manipulation of Sub-micron Particles in an Integrated Microfluidic Chip.

    PubMed

    Kim, Jisu; Shin, Jung H

    2016-09-22

    We demonstrate stable, free-space optical trapping and manipulation in an integrated microfluidic chip using counter-propagating beams. An inverted ridge-type waveguide made of SU8 is cut across by an open trench. The design of the waveguide provides low propagation losses and small divergence of the trapping beam upon emergence from the facet, and the trench designed to be deeper and wider than the optical mode enables full utilization of the optical power with an automatic alignment for counter-propagating beams in a trap volume away from all surfaces. After integration with polydimethylsiloxane (PDMS) microfluidic channel for particle delivery, 0.65 μm and 1 μm diameter polystyrene beads were trapped in free space in the trench, and manipulated to an arbitrary position between the waveguides with a resolution of < 100 nm. Comparison with numerical simulations confirm stable trapping of sub-micron particles, with a 10 kBT threshold power of less than 1 mW and a stiffness that can be 1 order of magnitude larger than that of comparable fiber-based trapping methods.

  17. Stable, Free-space Optical Trapping and Manipulation of Sub-micron Particles in an Integrated Microfluidic Chip

    NASA Astrophysics Data System (ADS)

    Kim, Jisu; Shin, Jung H.

    2016-09-01

    We demonstrate stable, free-space optical trapping and manipulation in an integrated microfluidic chip using counter-propagating beams. An inverted ridge-type waveguide made of SU8 is cut across by an open trench. The design of the waveguide provides low propagation losses and small divergence of the trapping beam upon emergence from the facet, and the trench designed to be deeper and wider than the optical mode enables full utilization of the optical power with an automatic alignment for counter-propagating beams in a trap volume away from all surfaces. After integration with polydimethylsiloxane (PDMS) microfluidic channel for particle delivery, 0.65 μm and 1 μm diameter polystyrene beads were trapped in free space in the trench, and manipulated to an arbitrary position between the waveguides with a resolution of < 100 nm. Comparison with numerical simulations confirm stable trapping of sub-micron particles, with a 10 kBT threshold power of less than 1 mW and a stiffness that can be 1 order of magnitude larger than that of comparable fiber-based trapping methods.

  18. Stable, Free-space Optical Trapping and Manipulation of Sub-micron Particles in an Integrated Microfluidic Chip

    PubMed Central

    Kim, Jisu; Shin, Jung H.

    2016-01-01

    We demonstrate stable, free-space optical trapping and manipulation in an integrated microfluidic chip using counter-propagating beams. An inverted ridge-type waveguide made of SU8 is cut across by an open trench. The design of the waveguide provides low propagation losses and small divergence of the trapping beam upon emergence from the facet, and the trench designed to be deeper and wider than the optical mode enables full utilization of the optical power with an automatic alignment for counter-propagating beams in a trap volume away from all surfaces. After integration with polydimethylsiloxane (PDMS) microfluidic channel for particle delivery, 0.65 μm and 1 μm diameter polystyrene beads were trapped in free space in the trench, and manipulated to an arbitrary position between the waveguides with a resolution of < 100 nm. Comparison with numerical simulations confirm stable trapping of sub-micron particles, with a 10 kBT threshold power of less than 1 mW and a stiffness that can be 1 order of magnitude larger than that of comparable fiber-based trapping methods. PMID:27653191

  19. An integrated microfluidic chip with 40 MHz lead-free transducer for fluid analysis.

    PubMed

    Lee, S T F; Lam, K H; Lei, L; Zhang, X M; Chan, H L W

    2011-02-01

    The design, fabrication, and evaluation of a high-frequency transducer made from lead-free piezoceramic for the application of microfluidic analysis is described. Barium strontium zirconate titanate [(Ba(0.95)Sr(0.05))(Zr(0.05)Ti(0.95))O(3), abbreviated as BSZT] ceramic has been chosen to be the active element of the transducer. The center frequency and bandwidth of this high-frequency ultrasound transducer have been measured to be 43 MHz and 56.1%, respectively. The transducer was integrated into a microfluidic channel and used to measure the sound velocity and attenuation of the liquid flowing in the channel. Results suggest that lead-free high-frequency transducers could be used for in situ analysis of property of the fluid flowing through the microfluidic system.

  20. Fabrication of microfluidic architectures for optimal flow rate and concentration measurement for lab on chip application

    NASA Astrophysics Data System (ADS)

    Adam, Tijjani; Hashim, U.

    2017-03-01

    Optimum flow in micro channel for sensing purpose is challenging. In this study, The optimizations of the fluid sample flows are made through the design and characterization of the novel microfluidics' architectures to achieve the optimal flow rate in the micro channels. The biocompatibility of the Polydimetylsiloxane (Sylgard 184 silicon elastomer) polymer used to fabricate the device offers avenue for the device to be implemented as the universal fluidic delivery system for bio-molecules sensing in various bio-medical applications. The study uses the following methodological approaches, designing a novel microfluidics' architectures by integrating the devices on a single 4 inches silicon substrate, fabricating the designed microfluidic devices using low-cost solution soft lithography technique, characterizing and validating the flow throughput of urine samples in the micro channels by generating pressure gradients through the devices' inlets. The characterization on the urine samples flow in the micro channels have witnessed the constant flow throughout the devices.

  1. Protein crystallization using microfluidic technologies based on valves, droplets, and SlipChip.

    PubMed

    Li, Liang; Ismagilov, Rustem F

    2010-01-01

    To obtain protein crystals, researchers must search for conditions in multidimensional chemical space. Empirically, thousands of crystallization experiments are carried out to screen various precipitants at multiple concentrations. Microfluidics can manipulate fluids on a nanoliter scale, and it affects crystallization twofold. First, it miniaturizes the experiments that can currently be done on a larger scale and enables crystallization of proteins that are available only in small amounts. Second, it offers unique experimental approaches that are difficult or impossible to implement on a larger scale. Ongoing development of microfluidic techniques and their integration with protein production, characterization, and in situ diffraction promises to accelerate the progress of structural biology.

  2. Microfluidic chip-based liquid-liquid extraction and preconcentration using a subnanoliter-droplet trapping technique.

    PubMed

    Chen, Hong; Fang, Qun; Yin, Xue-Feng; Fang, Zhao-Lun

    2005-07-01

    A robust and simple approach for microfluidic liquid-liquid (L-L) extraction at the subnanoliter-scale was developed for on-chip sample pretreatment. Organic solvent droplets of a few hundred pL were trapped within micro recesses fabricated in the channel walls of a microfabricated glass chip. L-L extraction was performed by delivering aqueous samples through the channel, with the sample stream continuously flowing adjacent to the droplets. The analytes in aqueous streams were enriched within the droplet with high preconcentration factors owing to both phase transfer and dissolution of organic solvent into the bypassing aqueous sample. An aqueous solution of butyl rhodamine B (BRB) and 1-hexanol were used, respectively, as sample and extractant to demonstrate the performance of the system. The fluorescence intensity of the dye extracted into the droplet was monitored in situ by LIF. The system proved to be an efficient means for achieving high enrichment factors of over 1000, with sample consumption of a few microL. Quantitative measurement of the extracted analyte was achieved with a linear response in the range 1 x 10(-9)-8 x 10(-7) M BRB. The precision of the measured fluorescence values for a 10(-7) M BRB standard with a 12.5 min preconcentration period was 6.6% RSD (n = 5).

  3. All-electronic droplet generation on-chip with real-time feedback control for EWOD digital microfluidics.

    PubMed

    Gong, Jian; Kim, Chang-Jin C J

    2008-06-01

    Electrowetting-on-dielectric (EWOD) actuation enables digital (or droplet) microfluidics where small packets of liquids are manipulated on a two-dimensional surface. Due to its mechanical simplicity and low energy consumption, EWOD holds particular promise for portable systems. To improve volume precision of the droplets, which is desired for quantitative applications such as biochemical assays, existing practices would require near-perfect device fabrication and operation conditions unless the droplets are generated under feedback control by an extra pump setup off of the chip. In this paper, we develop an all-electronic (i.e., no ancillary pumping) real-time feedback control of on-chip droplet generation. A fast voltage modulation, capacitance sensing, and discrete-time PID feedback controller are integrated on the operating electronic board. A significant improvement is obtained in the droplet volume uniformity, compared with an open loop control as well as the previous feedback control employing an external pump. Furthermore, this new capability empowers users to prescribe the droplet volume even below the previously considered minimum, allowing, for example, 1 : x (x < 1) mixing, in comparison to the previously considered n : m mixing (i.e., n and m unit droplets).

  4. A Microfluidic ExoSearch Chip for Multiplexed Exosome Detection Towards Blood-based Ovarian Cancer Diagnosis†

    PubMed Central

    Zhao, Zheng; Yang, Yang; Zeng, Yong; He, Mei

    2015-01-01

    Tumor-derived circulating exosomes, enriched with a group of tumor antigens, have been recognized as a promising biomarker source for cancer diagnosis via less invasive procedure. Quantitatively pinpointing exosome tumor markers is appealing, yet challenging. In this study, we developed a simple microfluidic approach (ExoSearch) which provides enriched preparation of blood plasma exosomes for in-situ, multiplexed detection using immunomagnetic beads. The ExosSearch chip offers robust, continuous-flow design for quantitative isolation and release of blood plasma exosomes in a wide range of preparation volumes (10 μL to 10 mL). We employed the ExoSearch chip for blood-based diagnosis of ovarian cancer by multiplexed measurement of three exosomal tumor markers (CA-125, EpCAM, CD24) using a training set of ovarian cancer patient plasma, which showed significant diagnostic power (a.u.c. = 1.0, p = 0.001) and was comparable with standard Bradford assay. This work provides an essentially needed platform for utilization of exosomes in clinical cancer diagnosis, as well as fundamental exosome research. PMID:26645590

  5. ALL-ELECTRONIC DROPLET GENERATION ON-CHIP WITH REAL-TIME FEEDBACK CONTROL FOR EWOD DIGITIAL MICROFLUIDICS

    PubMed Central

    Gong, Jian; Kim, Chang-Jin “CJ”

    2009-01-01

    Electrowetting-on-dielectric (EWOD) actuation enables digital (or droplet) microfluidics where small packets of liquids are manipulated on a two-dimensional surface. Due to its mechanical simplicity and low energy consumption, EWOD holds particular promise for portable systems. To improve volume precision of the droplets, which is desired for quantitative applications such as biochemical assays, existing practices would require near-perfect device fabricaion and operation conditions unless the droplets are generated under feedback control by an extra pump setup off of the chip. In this paper, we develop an all-electronic (i.e., no ancillary pumping) real-time feedback control of on-chip droplet generation. A fast voltage modulation, capacitance sensing, and discrete-time PID feedback controller are integrated on the operating electronic board. A significant improvement is obtained in the droplet volume uniformity, compared with an open loop control as well as the previous feedback control employing an external pump. Furthermore, this new capability empowers users to prescribe the droplet volume even below the previously considered minimum, allowing, for example, 1:x (x < 1) mixing, in comparison to the previously considered n:m mixing (i.e., n and m unit droplets). PMID:18497909

  6. A biomimetic microfluidic chip to study the circulation and mechanical retention of red blood cells in the spleen.

    PubMed

    Picot, Julien; Ndour, Papa Alioune; Lefevre, Sophie D; El Nemer, Wassim; Tawfik, Harvey; Galimand, Julie; Da Costa, Lydie; Ribeil, Jean-Antoine; de Montalembert, Mariane; Brousse, Valentine; Le Pioufle, Bruno; Buffet, Pierre; Le Van Kim, Caroline; Français, Olivier

    2015-04-01

    Red blood cells (RBCs) are deformable and flow through vessels narrower than their own size. Their deformability is most stringently challenged when they cross micrometer-wide slits in the spleen. In several inherited or acquired RBC disorders, blockade of small vessels by stiff RBCs can trigger organ damage, but a functional spleen is expected to clear these abnormal RBCs from the circulation before they induce such complications. We analyzed flow behavior of RBCs in a microfluidic chip that replicates the mechanical constraints imposed on RBCs as they cross the human spleen. Polymer microchannels obtained by soft lithography with a hydraulic diameter of 25 μm drove flow into mechanical filtering units where RBCs flew either slowly through 5- to 2-μm-wide slits or rapidly along 10-μm-wide channels, these parallel paths mimicking the splenic microcirculation. Stiff heated RBCs accumulated in narrow slits seven times more frequently than normal RBCs infused simultaneously. Stage-dependent retention of Plasmodium falciparum-infected RBCs was also observed in these slits. We also analyzed RBCs from patients with hereditary spherocytosis and observed retention for those having the most altered mechanical properties as determined by ektacytometry. Thus, in keeping with previous observations in vivo and ex vivo, the chip successfully discriminated poorly deformable RBCs based on their distinct mechanical properties and on the intensity of the cell alteration. Applications to the exploration of the pathogenesis of malaria, hereditary spherocytosis, sickle cell disease and other RBC disorders are envisioned.

  7. Hands-off preparation of monodisperse emulsion droplets using a poly(dimethylsiloxane) microfluidic chip for droplet digital PCR.

    PubMed

    Tanaka, Hironari; Yamamoto, Shunsuke; Nakamura, Arichika; Nakashoji, Yuta; Okura, Naoaki; Nakamoto, Norimitsu; Tsukagoshi, Kazuhiko; Hashimoto, Masahiko

    2015-04-21

    A fully autonomous method of creating highly monodispersed emulsion droplets with a low sample dead volume was realized using a degassed poly(dimethylsiloxane) (PDMS) microfluidic chip possessing a simple T-junction channel geometry with two inlet reservoirs for oil and water to be loaded and one outlet reservoir for the collection of generated droplets. Autonomous transport of oil and water phases in the channel was executed by permeation of air confined inside the outlet reservoir into the degassed PDMS. The only operation required for droplet creation was simple pipetting of oil and aqueous solutions into the inlet reservoirs. Long-lasting fluid transport in the current system enabled us to create ca. 51,000 monodispersed droplets (with a coefficient of variation of <3% for the droplet diameter) in 80 min with a maximum droplet generation rate of ca. 12 Hz using a PDMS chip that had been degassed overnight. With multiple time-course measurements, the reproducibility in the current method of droplet preparation was confirmed, with tunable droplet sizes achieved simply by changing the cross-sectional dimensions of the microchannel. Furthermore, it was verified that the resultant droplets could serve as microreactors for digital polymerase chain reactions. This hands-free technique for preparing monodispersed droplets in a very facile and inexpensive fashion is intended for, but not limited to, bioanalytical applications and is also applicable to material syntheses.

  8. Application of microfluidic chip with integrated optics for electrophoretic separations of proteins.

    PubMed

    Vieillard, Julien; Mazurczyk, Radoslaw; Morin, Christophe; Hannes, Benjamin; Chevolot, Yann; Desbène, Paul-Louis; Krawczyk, Stanislas

    2007-01-15

    This paper describes the fabrication, the characterization and the applications of a capillary electrophoresis microchip. This hybrid device (glass/PDMS) features channels and optical waveguides integrated in one common substrate. It can be used for electrophoretic separation and fluorimetric detection of molecules. The microfluidic performance of the device is demonstrated by capillary zone and gel electrophoresis of proteins.

  9. Microfluidics Integrated Biosensors: A Leading Technology towards Lab-on-a-Chip and Sensing Applications.

    PubMed

    Luka, George; Ahmadi, Ali; Najjaran, Homayoun; Alocilja, Evangelyn; DeRosa, Maria; Wolthers, Kirsten; Malki, Ahmed; Aziz, Hassan; Althani, Asmaa; Hoorfar, Mina

    2015-12-01

    A biosensor can be defined as a compact analytical device or unit incorporating a biological or biologically derived sensitive recognition element immobilized on a physicochemical transducer to measure one or more analytes. Microfluidic systems, on the other hand, provide throughput processing, enhance transport for controlling the flow conditions, increase the mixing rate of different reagents, reduce sample and reagents volume (down to nanoliter), increase sensitivity of detection, and utilize the same platform for both sample preparation and detection. In view of these advantages, the integration of microfluidic and biosensor technologies provides the ability to merge chemical and biological components into a single platform and offers new opportunities for future biosensing applications including portability, disposability, real-time detection, unprecedented accuracies, and simultaneous analysis of different analytes in a single device. This review aims at representing advances and achievements in the field of microfluidic-based biosensing. The review also presents examples extracted from the literature to demonstrate the advantages of merging microfluidic and biosensing technologies and illustrate the versatility that such integration promises in the future biosensing for emerging areas of biological engineering, biomedical studies, point-of-care diagnostics, environmental monitoring, and precision agriculture.

  10. Development and Fabrication of Nanoporous Silicon-based Bioreactors within a Microfluidic Chip

    SciTech Connect

    Siuti, Piro; Choi, Chang Kyoung; Doktycz, Mitchel John; Retterer, Scott T

    2010-01-01

    Multi-scale lithography and cryogenic deep reactive ion etching techniques were used to create ensembles of nanoporous, picoliter volume, reaction vessels within a microfluidic system. The fabrication of these vessels is described and how this process can be used to tailor vessel porosity by controlling the width of slits that constitute the vessel pores is demonstrated. Control of pore size allows the containment of nucleic acids and enzymes that are the foundation of biochemical reaction systems, while allowing smaller reaction constituents to traverse the container membrane and continuously supply the reaction. In this work, a 5.4kB DNA plasmid was retained within the reaction vessels and labeled under microfluidic control with ethidium bromide as an initial proof-of-principle. Subsequently, a coupled enzyme reaction, in which glucose oxidase and horseradish peroxidase were contained and fed with a substrate solution of glucose and Amplex Red to produce fluorescent Resorufin, was carried out under microfluidic control and monitored using fluorescent microscopy. The fabrication techniques presented are broadly applicable and can be adapted to produce devices in which a variety of high aspect ratio, nanoporous silicon structures can be integrated within a microfluidic network. The devices shown here are amenable to being scaled in number and organized to implement more complex reaction systems for applications in sensing and production of biologically based therapeutics as well as fundamental studies of biological reaction systems.

  11. Microfluidics Integrated Biosensors: A Leading Technology towards Lab-on-a-Chip and Sensing Applications

    PubMed Central

    Luka, George; Ahmadi, Ali; Najjaran, Homayoun; Alocilja, Evangelyn; DeRosa, Maria; Wolthers, Kirsten; Malki, Ahmed; Aziz, Hassan; Althani, Asmaa; Hoorfar, Mina

    2015-01-01

    A biosensor can be defined as a compact analytical device or unit incorporating a biological or biologically derived sensitive recognition element immobilized on a physicochemical transducer to measure one or more analytes. Microfluidic systems, on the other hand, provide throughput processing, enhance transport for controlling the flow conditions, increase the mixing rate of different reagents, reduce sample and reagents volume (down to nanoliter), increase sensitivity of detection, and utilize the same platform for both sample preparation and detection. In view of these advantages, the integration of microfluidic and biosensor technologies provides the ability to merge chemical and biological components into a single platform and offers new opportunities for future biosensing applications including portability, disposability, real-time detection, unprecedented accuracies, and simultaneous analysis of different analytes in a single device. This review aims at representing advances and achievements in the field of microfluidic-based biosensing. The review also presents examples extracted from the literature to demonstrate the advantages of merging microfluidic and biosensing technologies and illustrate the versatility that such integration promises in the future biosensing for emerging areas of biological engineering, biomedical studies, point-of-care diagnostics, environmental monitoring, and precision agriculture. PMID:26633409

  12. High-throughput nanoliter sample introduction microfluidic chip-based flow injection analysis system with gravity-driven flows.

    PubMed

    Du, Wen-Bin; Fang, Qun; He, Qiao-Hong; Fang, Zhao-Lun

    2005-03-01

    In this work, a simple, robust, and automated microfluidic chip-based FIA system with gravity-driven flows and liquid-core waveguide (LCW) spectrometric detection was developed. The high-throughput sample introduction system was composed of a capillary sampling probe and an array of horizontally positioned microsample vials with a slot fabricated on the bottom of each vial. FI sample loading and injection were performed by linearly moving the array of vials filled alternately with 50-microL samples and carrier, allowing the probe inlet to enter the solutions in the vials through the slots sequentially and the sample and carrier solution to be introduced into the chip driven by gravity. The performance of the system was demonstrated using the complexation of o-phenanthroline with Fe(II) as a model reaction. A 20-mm-long Teflon AF 2400 capillary (50-microm i.d., 375-microm o.d.) was connected to the chip to function as a LCW detection flow cell with a cell volume of 40 nL and effective path length of 1.7 cm. Linear absorbance response was obtained in the range of 1.0-100 microM Fe(II) (r2=0.9967), and a good reproducibility of 0.6% RSD (n=18) was achieved. The sensitivity was comparable with that obtained using conventional FIA systems, which typically consume 10,000-fold more sample. The highest sampling throughput of 1000 h-1 was obtained by using injection times of 0.08 and 3.4 s for sample and carrier solution, respectively, with a sample consumption of only 0.6 nL for each cycle.

  13. Design and Construction of a Multi-Organ Microfluidic Chip Mimicking the in vivo Microenvironment of Lung Cancer Metastasis.

    PubMed

    Xu, Zhiyun; Li, Encheng; Guo, Zhe; Yu, Ruofei; Hao, Hualong; Xu, Yitong; Sun, Zhao; Li, Xiancheng; Lyu, Jianxin; Wang, Qi

    2016-10-05

    Metastasis is a complex pathophysiological process. As the main cause of cancer mortality in humans it represents a serious challenge to both basic researchers and clinicians. Here we report the design and construction of a multi-organ microfluidic chip that closely mimics the in vivo microenvironment of lung cancer metastasis. This multi-organs-on-a-chip includes an upstream "lung" and three downstream "distant organs", with three polydimethylsiloxane (PDMS) layers and two thin PDMS microporous membranes bonded to form three parallel microchannels. Bronchial epithelial, lung cancer, microvascular endothelial, mononuclear, and fibroblast cells were grown separated by the biomembrane in upstream "lung", while astrocytes, osteocytes, and hepatocytes were grown in distant chambers, to mimic lung cancer cell metastasis to the brain, bone, and liver. After culture in this system, lung cancer cells formed a "tumor mass", showed epithelial-mesenchymal transition (with altered expression of E-cadherin, N-cadherin, Snail1, and Snail2) and invasive capacity. A549 cells co-cultured with astrocytes overexpressed CXCR4 protein, indicating damage of astrocytes after cancer cell metastasis to the brain. Osteocytes overexpressed RANKL protein indicates damage of osteocytes after cancer cell metastasis to the bone, and hepatocytes overexpressed AFP protein indicates damage to hepatocytes after cancer cell metastasis to the liver. Finally, in vivo imaging of cancer growth and metastasis in a nude mice model validated the performance of metastasis in the organs-on-chip system. This system provides a useful tool to mimic the in vivo microenvironment of cancer metastasis and to investigate cell-cell interactions during metastasis.

  14. Crack-free direct-writing on glass using a low-power UV laser in the manufacture of a microfluidic chip

    NASA Astrophysics Data System (ADS)

    Cheng, Ji-Yen; Yen, Meng-Hua; Wei, Cheng-Wey; Chuang, Yung-Chuan; Young, Tai-Horng

    2005-06-01

    Glass is an excellent material for use as a microfluidic chip substrate because it has great chemical and thermal stability. This work describes a flexible platform for the rapid prototyping of microfluidic chips fabricated from glass. A debris-free laser direct-writing technology that requires no photomask generation is developed. A 266 nm laser with a high repetition rate is employed in laser-induced backside wet etching (LIBWE) for glass machining. A microfluidic pattern is designed using computer drawing software and then automatically translated into computer numerical control motion so that the microtrench is directly fabricated on the glass chip. The overall machining speed can be increased by increasing the repetition rate to ~6 kHz. Without a clean room facility or the highly corrosive acid, HF, the overall development time is within hours. Trenches with complex structures that are hard to fabricate by photolithography were easily produced by laser direct-writing. An integrated microreactor/concentrator is demonstrated. The crack-free and debris-free surface was characterized by SEM and a surface profiler. Various effective etching chemicals for the LIBWE process were investigated to understand the etching mechanism. The minimal laser power used for glass etching was approximately 20 mW for a 6 µm wide microtrench. Several new compounds have been demonstrated to be effective in ablation. The etch threshold is minimum and does not decrease further as the unit length absorbance increases above 8000 in acetone solution.

  15. Designing Knowledge of The PPC with Semantic Network

    NASA Astrophysics Data System (ADS)

    Fajar Santoso, Ari; Supriana, Iping; Surendro, Kridanto

    2017-01-01

    A manufacturing process is a very complex activity, which involves a variety of knowledge, such as the product design and description, manufacturing operations, tools, machines, and the relationships between these entities. Production planning control (PPC) as dominant part of manufacturing process has a very important role in determining the measures taken by the management of company. The PPC construction is an interactive process that requires the collaboration of both ICT and manufacture experts. Building PPC process can be used as a basis for making knowledge in manufacturing domain that will be used in manufacturing intelligent system. This paper describes about designing knowledge with semantic network in the production scheduling.

  16. Fit-to-Flow (F2F) interconnects: universal reversible adhesive-free microfluidic adaptors for lab-on-a-chip systems.

    PubMed

    Chen, Arnold; Pan, Tingrui

    2011-02-21

    World-to-chip (macro-to-micro) interface continues to be one of the most complicated, ineffective, and unreliable components in the development of emerging lab-on-a-chip systems involving integrated microfluidic operations. A number of irreversible (e.g., adhesive gluing) and reversible techniques (e.g., press fitting) have attempted to provide dedicated fluidic passage from standard tubing to miniature on-chip devices, none of which completely addresses the above concerns. In this paper, we present standardized adhesive-free microfluidic adaptors, referred to as Fit-to-Flow (F2F) Interconnects, to achieve reliable hermetic seal, high-density tube packing, self-aligned plug-in, reworkable connectivity, straightforward scalability and expandability, and applicability to broad lab-on-a-chip platforms; analogous to the modular plug-and-play USB architecture employed in modern electronics. Specifically, two distinct physical packaging mechanisms are applied, with one utilizing induced tensile stress in elastomeric socket to establish reversible seal and the other using negative pressure to provide on demand vacuum shield, both of which can be adapted to a variety of experimental configurations. The non-leaking performance (up to 336 kPa) along with high tube-packing density (of 1 tube/mm(2)) and accurate self-guided alignment (of 10 μm) have been characterized. In addition, a 3D microfluidic mixer and a 6-level chemical gradient generator paired with the corresponding F2F Interconnects have been devised to illustrate the applicability of the universal fluidic connections to classic lab-on-a-chip operations.

  17. Integration of dialysis membranes into a poly(dimethylsiloxane) microfluidic chip for isoelectric focusing of proteins using whole-channel imaging detection.

    PubMed

    Ou, Junjie; Glawdel, Tomasz; Samy, Razim; Wang, Shuwen; Liu, Zhen; Ren, Carolyn L; Pawliszyn, Janusz

    2008-10-01

    A poly(dimethylsiloxane) microfluidic chip-based cartridge is developed and reported here for protein analysis using isoelectic focusing (IEF)-whole-channel imaging detection (WCID) technology. In this design, commercial dialysis membranes are integrated to separate electrolytes and samples and to reduce undesired pressure-driven flow. Fused-silica capillaries are also incorporated in this design for sample injection and channel surface preconditioning. This structure is equivalent to that of a commercial fused-silica capillary-based cartridge for adapting to an IEF analyzer (iCE280 analyzer) to perform IEF-WCID. The successful integration of dialysis membranes into a microfluidic chip significantly improves IEF repeatability by eliminating undesired pressure-driven hydrodynamics and also makes sample injection much easier than that using the first-generation chip as reported recently. In this study, two microfluidic chips with a 100-microm-high, 100-microm-wide and a 200-microm-high, 50-microm-wide microchannel, respectively, were applied for qualitative and quantitative analysis of proteins. The mixture containing six pI markers with a pH range of 3-10 was successfully separated using IEF-WCID. The pH gradient exhibited a good linearity by plotting the pI value versus peak position, and the correlation coefficient reached 0.9994 and 0.9995 separately for the two chips. The separation of more complicated human hemoglobin control sample containing HbA, HbF, HbS, and HbC was also achieved. Additionally, for the quantitative analysis, a good linearity of IEF peak value versus myoglobin concentration in the range of 20-100 microg/mL was obtained.

  18. Retina-on-a-chip: a microfluidic platform for point access signaling studies

    PubMed Central

    Dodson, Kirsten H.; Echevarria, Franklin D.; Li, Deyu; Sappington, Rebecca M.; Edd, Jon F.

    2016-01-01

    We report on a microfluidic platform for culture of whole organs or tissue slices with the capability of point access reagent delivery to probe the transport of signaling events. Whole mice retina were maintained for multiple days with negative pressure applied to tightly but gently bind the bottom of the retina to a thin poly-(dimethylsiloxane) membrane, through which twelve 100 μm diameter through-holes served as fluidic access points. Staining with toluidine blue, transport of locally applied cholera toxin beta, and transient response to lipopolysaccharide in the retina demonstrated the capability of the microfluidic platform. The point access fluidic delivery capability could enable new assays in the study of various kinds of excised tissues, including retina. PMID:26559199

  19. Raman spectroscopy compatible PDMS droplet microfluidic culture and analysis platform towards on-chip lipidomics.

    PubMed

    Kim, Hyun Soo; Waqued, Sergio C; Nodurft, Dawson T; Devarenne, Timothy P; Yakovlev, Vladislav V; Han, Arum

    2017-04-07

    Lipids produced by microalgae are viewed as a potential renewable alternative to fossil fuels, however, significant improvements in productivity are required for microalgal biofuels to become economically feasible. Here we present a method that allows for the use of Raman spectroscopy with poly(dimethylsiloxane) (PDMS) droplet microfluidic devices, which not only overcomes the high Raman background of PDMS, but also achieves pairing of the high-throughput single-cell resolution advantages of droplet microfluidics with the direct, chemically specific, label-free, and non-destructive nature of Raman spectroscopy. The platform was successfully utilized for in situ characterization of microalgal lipid production over time within droplets, paving the way towards high-throughput microalgal lipidomics assays.

  20. Retina-on-a-chip: a microfluidic platform for point access signaling studies.

    PubMed

    Dodson, Kirsten H; Echevarria, Franklin D; Li, Deyu; Sappington, Rebecca M; Edd, Jon F

    2015-12-01

    We report on a microfluidic platform for culture of whole organs or tissue slices with the capability of point access reagent delivery to probe the transport of signaling events. Whole mice retina were maintained for multiple days with negative pressure applied to tightly but gently bind the bottom of the retina to a thin poly-(dimethylsiloxane) membrane, through which twelve 100 μm diameter through-holes served as fluidic access points. Staining with toluidine blue, transport of locally applied cholera toxin beta, and transient response to lipopolysaccharide in the retina demonstrated the capability of the microfluidic platform. The point access fluidic delivery capability could enable new assays in the study of various kinds of excised tissues, including retina.

  1. Diode laser bonding of planar microfluidic devices, MOEMS, bioMEMS, diagnostic chips, and microarrays

    NASA Astrophysics Data System (ADS)

    Chen, Jie-Wei; Zybko, Jerry M.

    2005-01-01

    The assembly of plastic microfluidic devices, MOEMS and microarrays requiring high positioning and welding accuracy in the micrometer range, has been successfully achieved using a new technology based on laser transmission welding combined with a photolithographic mask technique. This paper reviews a laser assembly platform for the joining of microfluidic plastic parts with its main related process characteristics and its potential for low-cost and high volume manufacturing. The system consists of a of diode laser with a mask and an automated alignment function to generate micro welding seams with freely definable geometries. A fully automated mask alignment system with a resolution of < 2 &mum and a precise, noncontact energy input allows a fast welding of micro structured plastic parts with high reproducibility and excellent welding quality.

  2. The study of mixing of reagents within a droplet in various designs of microfluidic chip

    NASA Astrophysics Data System (ADS)

    Filatov, N. A.; Belousov, K. I.; Bukatin, A. S.; Kukhtevich, I. V.; Evstrapov, A. A.

    2016-08-01

    The aim of this work was to determine a design of a microfluidic droplet generator which provides the best mixing of two components within a droplet during its formation. Simulation of droplets’ formation in various designs of microfluidic generators was performed. Dependences of a mixing index from a capillary number and a viscosity of continuous phase were obtained for each design. The mixing index was determined directly after the formation of the droplet. It was found that the small capillary numbers correspond to the lower mixing index due to increase of the droplets diameter. The high viscosity of the continuous phase provides vortex flows in the area of droplet formation which leads to an increase in the mixing index for asymmetric designs, but has no significant effect on the mixing in conventional symmetric flow focusing. So for the asymmetric droplet generator designs the mixing index increases up to 1.5 as compared with the conventional flow focusing designs.

  3. A multichannel acoustically driven microfluidic chip to study particle-cell interactions

    PubMed Central

    Wang, Xue-Yan; Fillafer, Christian; Pichl, Clara; Deinhammer, Stephanie; Hofer-Warbinek, Renate; Wirth, Michael; Gabor, Franz

    2013-01-01

    Microfluidic devices have emerged as important tools for experimental physiology. They allow to study the effects of hydrodynamic flow on physiological and pathophysiological processes, e.g., in the circulatory system of the body. Such dynamic in vitro test systems are essential in order to address fundamental problems in drug delivery and targeted imaging, such as the binding of particles to cells under flow. In the present work an acoustically driven microfluidic platform is presented in which four miniature flow channels can be operated in parallel at distinct flow velocities with only slight inter-experimental variations. The device can accommodate various channel architectures and is fully compatible with cell culture as well as microscopy. Moreover, the flow channels can be readily separated from the surface acoustic wave pumps and subsequently channel-associated luminescence, absorbance, and/or fluorescence can be determined with a standard microplate reader. In order to create artificial blood vessels, different coatings were evaluated for the cultivation of endothelial cells in the microchannels. It was found that 0.01% fibronectin is the most suitable coating for growth of endothelial monolayers. Finally, the microfluidic system was used to study the binding of 1 μm polystyrene microspheres to three different types of endothelial cell monolayers (HUVEC, HUVECtert, HMEC-1) at different average shear rates. It demonstrated that average shear rates between 0.5 s−1 and 2.25 s−1 exert no significant effect on cytoadhesion of particles to all three types of endothelial monolayers. In conclusion, the multichannel microfluidic platform is a promising device to study the impact of hydrodynamic forces on cell physiology and binding of drug carriers to endothelium. PMID:24404060

  4. Microcirculation-on-a-Chip: A Microfluidic Platform for Assaying Blood- and Lymphatic-Vessel Permeability

    PubMed Central

    Sato, Miwa; Sasaki, Naoki; Ato, Manabu; Hirakawa, Satoshi; Sato, Kiichi; Sato, Kae

    2015-01-01

    We developed a microfluidic model of microcirculation containing both blood and lymphatic vessels for examining vascular permeability. The designed microfluidic device harbors upper and lower channels that are partly aligned and are separated by a porous membrane, and on this membrane, blood vascular endothelial cells (BECs) and lymphatic endothelial cells (LECs) were cocultured back-to-back. At cell-cell junctions of both BECs and LECs, claudin-5 and VE-cadherin were detected. The permeability coefficient measured here was lower than the value reported for isolated mammalian venules. Moreover, our results showed that the flow culture established in the device promoted the formation of endothelial cell-cell junctions, and that treatment with histamine, an inflammation-promoting substance, induced changes in the localization of tight and adherens junction-associated proteins and an increase in vascular permeability in the microdevice. These findings indicated that both BECs and LECs appeared to retain their functions in the microfluidic coculture platform. Using this microcirculation device, the vascular damage induced by habu snake venom was successfully assayed, and the assay time was reduced from 24 h to 30 min. This is the first report of a microcirculation model in which BECs and LECs were cocultured. Because the micromodel includes lymphatic vessels in addition to blood vessels, the model can be used to evaluate both vascular permeability and lymphatic return rate. PMID:26332321

  5. Drop mass transfer in a microfluidic chip compared to a centrifugal contactor

    DOE PAGES

    Nemer, Martin B.; Roberts, Christine C.; Hughes, Lindsey G.; ...

    2014-06-13

    A model system was developed for enabling a multiscale understanding of centrifugal-contactor liquid–liquid extraction.The system consisted of Nd(III) + xylenol orange in the aqueous phase buffered to pH =5.5 by KHP, and dodecane + thenoyltrifluroroacetone (HTTA) + tributyphosphate (TBP) in the organic phase. Diffusion constants were measured for neodymium in both the organic and aqueous phases, and the Nd(III) partition coefficients were measured at various HTTA and TBP concentrations. A microfluidic channel was used as a high-shear model environment to observe mass-transfer on a droplet scale with xylenol orange as the aqueous-phase metal indicator; mass-transfer rates were measured quantitatively inmore » both diffusion and reaction limited regimes on the droplet scale. Lastly, the microfluidic results were comparable to observations made for the same system in a laboratory scale liquid–liquid centrifugal contactor, indicating that single drop microfluidic experiments can provide information on mass transfer in complicated flows and geometries.« less

  6. Integrated Microfluidic Membrane Transistor Utilizing Chemical Information for On-Chip Flow Control

    PubMed Central

    Frank, Philipp; Schreiter, Joerg; Haefner, Sebastian; Paschew, Georgi; Voigt, Andreas; Richter, Andreas

    2016-01-01

    Microfluidics is a great enabling technology for biology, biotechnology, chemistry and general life sciences. Despite many promising predictions of its progress, microfluidics has not reached its full potential yet. To unleash this potential, we propose the use of intrinsically active hydrogels, which work as sensors and actuators at the same time, in microfluidic channel networks. These materials transfer a chemical input signal such as a substance concentration into a mechanical output. This way chemical information is processed and analyzed on the spot without the need for an external control unit. Inspired by the development electronics, our approach focuses on the development of single transistor-like components, which have the potential to be used in an integrated circuit technology. Here, we present membrane isolated chemical volume phase transition transistor (MIS-CVPT). The device is characterized in terms of the flow rate from source to drain, depending on the chemical concentration in the control channel, the source-drain pressure drop and the operating temperature. PMID:27571209

  7. Drop mass transfer in a microfluidic chip compared to a centrifugal contactor

    SciTech Connect

    Nemer, Martin B.; Roberts, Christine C.; Hughes, Lindsey G.; Wyatt, Nicholas B.; Brooks, Carlton F.; Rao, Rekha

    2014-06-13

    A model system was developed for enabling a multiscale understanding of centrifugal-contactor liquid–liquid extraction.The system consisted of Nd(III) + xylenol orange in the aqueous phase buffered to pH =5.5 by KHP, and dodecane + thenoyltrifluroroacetone (HTTA) + tributyphosphate (TBP) in the organic phase. Diffusion constants were measured for neodymium in both the organic and aqueous phases, and the Nd(III) partition coefficients were measured at various HTTA and TBP concentrations. A microfluidic channel was used as a high-shear model environment to observe mass-transfer on a droplet scale with xylenol orange as the aqueous-phase metal indicator; mass-transfer rates were measured quantitatively in both diffusion and reaction limited regimes on the droplet scale. Lastly, the microfluidic results were comparable to observations made for the same system in a laboratory scale liquid–liquid centrifugal contactor, indicating that single drop microfluidic experiments can provide information on mass transfer in complicated flows and geometries.

  8. High-throughput on-chip in vivo neural regeneration studies using femtosecond laser nano-surgery and microfluidics

    NASA Astrophysics Data System (ADS)

    Rohde, Christopher B.; Zeng, Fei; Gilleland, Cody; Samara, Chrysanthi; Yanik, Mehmet F.

    2009-02-01

    In recent years, the advantages of using small invertebrate animals as model systems for human disease have become increasingly apparent and have resulted in three Nobel Prizes in medicine or chemistry during the last six years for studies conducted on the nematode Caenorhabditis elegans (C. elegans). The availability of a wide array of species-specific genetic techniques, along with the transparency of the worm and its ability to grow in minute volumes make C. elegans an extremely powerful model organism. We present a suite of technologies for complex high-throughput whole-animal genetic and drug screens. We demonstrate a high-speed microfluidic sorter that can isolate and immobilize C. elegans in a well-defined geometry, an integrated chip containing individually addressable screening chambers for incubation and exposure of individual animals to biochemical compounds, and a device for delivery of compound libraries in standard multiwell plates to microfluidic devices. The immobilization stability obtained by these devices is comparable to that of chemical anesthesia and the immobilization process does not affect lifespan, progeny production, or other aspects of animal health. The high-stability enables the use of a variety of key optical techniques. We use this to demonstrate femtosecond-laser nanosurgery and three-dimensional multiphoton microscopy. Used alone or in various combinations these devices facilitate a variety of high-throughput assays using whole animals, including mutagenesis and RNAi and drug screens at subcellular resolution, as well as high-throughput high-precision manipulations such as femtosecond-laser nanosurgery for large-scale in vivo neural degeneration and regeneration studies.

  9. Mimicking liver sinusoidal structures and functions using a 3D-configured microfluidic chip.

    PubMed

    Du, Yu; Li, Ning; Yang, Hao; Luo, Chunhua; Gong, Yixin; Tong, Chunfang; Gao, Yuxin; Lü, Shouqin; Long, Mian

    2017-02-28

    Physiologically, four major types of hepatic cells - the liver sinusoidal endothelial cells, Kupffer cells, hepatic stellate cells, and hepatocytes - reside inside liver sinusoids and interact with flowing peripheral cells under blood flow. It is hard to mimic an in vivo liver sinusoid due to its complex multiple cell-cell interactions, spatiotemporal construction, and mechanical microenvironment. Here we developed an in vitro liver sinusoid chip by integrating the four types of primary murine hepatic cells into two adjacent fluid channels separated by a porous permeable membrane, replicating liver's key structures and configurations. Each type of cells was identified with its respective markers, and the assembled chip presented the liver-specific unique morphology of fenestration. The flow field in the liver chip was quantitatively analyzed by computational fluid dynamics simulations and particle tracking visualization tests. Intriguingly, co-culture and shear flow enhance albumin secretion independently or cooperatively, while shear flow alone enhances HGF production and CYP450 metabolism. Under lipopolysaccharide (LPS) stimulations, the hepatic cell co-culture facilitated neutrophil recruitment in the liver chip. Thus, this 3D-configured in vitro liver chip integrates the two key factors of shear flow and the four types of primary hepatic cells to replicate key structures, hepatic functions, and primary immune responses and provides a new in vitro model to investigate the short-duration hepatic cellular interactions under a microenvironment mimicking the physiology of a liver.

  10. The feasibility of liquid sample microanalysis using polydimethylsiloxane microfluidic chips with in-channel and in-port laser-induced breakdown spectroscopy detection

    NASA Astrophysics Data System (ADS)

    Metzinger, Anikó; Nagy, Andrea; Gáspár, Attila; Márton, Zsuzsanna; Kovács-Széles, Éva; Galbács, Gábor

    2016-12-01

    This study describes the direct interfacing of polydimethylsiloxane (PDMS) microfluidic chips with laser-induced breakdown spectroscopy (LIBS) detection. The changes induced in the PDMS material by nanosecond laser ablation are briefly documented by using optical microscopy and scanning profilometry. The main part of the study focuses on the solution of technical and analytical problems of coupling single-pulse LIBS detection with PDMS microfluidic chips in order to assess the feasibility and performance of the concept of creating a lab-on-a-chip device with LIBS detection (LOC-LIBS). Multiple optical and sample presentation schemes including in-channel and in-port detection were tested, but it was found that LOC-LIBS is only viable and practical with in-port detection outside the chip. It was shown that LOC-LIBS in this configuration is capable of the trace speciation analysis of chromium using as little as 0.5 μL solution volume. The achieved absolute limit of detection was 2 ng.

  11. Microfluidic chemical reaction circuits

    SciTech Connect

    Lee, Chung-cheng; Sui, Guodong; Elizarov, Arkadij; Kolb, Hartmuth C; Huang, Jiang; Heath, James R; Phelps, Michael E; Quake, Stephen R; Tseng, Hsian-rong; Wyatt, Paul; Daridon, Antoine

    2012-06-26

    New microfluidic devices, useful for carrying out chemical reactions, are provided. The devices are adapted for on-chip solvent exchange, chemical processes requiring multiple chemical reactions, and rapid concentration of reagents.

  12. Label-free isolation of a prostate cancer cell among blood cells and the single-cell measurement of drug accumulation using an integrated microfluidic chip

    PubMed Central

    Khamenehfar, A.; Beischlag, T. V.; Russell, P. J.; Ling, M. T. P.; Nelson, C.; Li, P. C. H.

    2015-01-01

    Circulating tumor cells (CTCs) are found in the blood of patients with cancer. Although these cells are rare, they can provide useful information for chemotherapy. However, isolation of these rare cells from blood is technically challenging because they are small in numbers. An integrated microfluidic chip, dubbed CTC chip, was designed and fabricated for conducting tumor cell isolation. As CTCs usually show multidrug resistance (MDR), the effect of MDR inhibitors on chemotherapeutic drug accumulation in the isolated single tumor cell is measured. As a model of CTC isolation, human prostate cancer cells were mixed with mouse blood cells and the label-free isolation of the tumor cells was conducted based on cell size difference. The major advantages of the CTC chip are the ability for fast cell isolation, followed by multiple rounds of single-cell measurements, suggesting a potential assay for detecting the drug responses based on the liquid biopsy of cancer patients. PMID:26594265

  13. High performance magnesium anode in paper-based microfluidic battery, powering on-chip fluorescence assay

    PubMed Central

    Koo, Youngmi; Sankar, Jagannathan; Yun, Yeoheung

    2014-01-01

    A high power density and long-lasting stable/disposable magnesium battery anode was explored for a paper-based fluidic battery to power on-chip functions of various Point of Care (POC) devices. The single galvanic cell with magnesium foil anode and silver foil cathode in Origami cellulose chip provided open circuit potential, 2.2 V, and power density, 3.0 mW/cm2. A paper-based fluidic galvanic cell was operated with one drop of water (80 μl) and continued to run until it was dry. To prove the concept about powering on-chip POC devices, two-serial galvanic cells are developed and incorporated with a UV-light emitting diode (λ = 365 nm) and fluorescence assay for alkaline phosphatase reaction. Further, detection using smart phones was performed for quantitative measurement of fluorescent density. To conclude, a magnesium-based fluidic battery paper chip was extremely low-cost, required minute sample volumes, was easy to dispose of, light weight, easy to stack, store and transport, easy to fabricate, scalable, and has faster analysis times. PMID:25332741

  14. Acoustic resonances in microfluidic chips: full-image micro-PIV experiments and numerical simulations.

    PubMed

    Hagsäter, S M; Jensen, T Glasdam; Bruus, H; Kutter, J P

    2007-10-01

    We show that full-image micro-PIV analysis in combination with images of transient particle motion is a powerful tool for experimental studies of acoustic radiation forces and acoustic streaming in microfluidic chambers under piezo-actuation in the MHz range. The measured steady-state motion of both large 5 microm and small 1 microm particles can be understood in terms of the acoustic eigenmodes or standing ultra-sound waves in the given experimental microsystems. This interpretation is supported by numerical solutions of the corresponding acoustic wave equation.

  15. Plant-in-chip: Microfluidic system for studying root growth and pathogenic interactions in Arabidopsis

    NASA Astrophysics Data System (ADS)

    Parashar, Archana; Pandey, Santosh

    2011-06-01

    We report a microfluidic platform for the hydroponic growth of Arabidopsis plants with high-resolution visualization of root development and root-pathogen interactions. The platform comprises a set of parallel microchannels with individual input/output ports where 1-day old germinated seedlings are initially placed. Under optimum conditions, a root system grows in each microchannel and its images are recorded over a 198-h period. Different concentrations of plant growth media show different root growth characteristics. Later, the developed roots are inoculated with two plant pathogens (nematodes and zoospores) and their physicochemical interactions with the live root systems are observed.

  16. Size-based cell sorting with a resistive pulse sensor and an electromagnetic pump in a microfluidic chip.

    PubMed

    Song, Yongxin; Li, Mengqi; Pan, Xinxiang; Wang, Qi; Li, Dongqing

    2015-02-01

    An electrokinetic microfluidic chip is developed to detect and sort target cells by size from human blood samples. Target-cell detection is achieved by a differential resistive pulse sensor (RPS) based on the size difference between the target cell and other cells. Once a target cell is detected, the detected RPS signal will automatically actuate an electromagnetic pump built in a microchannel to push the target cell into a collecting channel. This method was applied to automatically detect and sort A549 cells and T-lymphocytes from a peripheral fingertip blood sample. The viability of A549 cells sorted in the collecting well was verified by Hoechst33342 and propidium iodide staining. The results show that as many as 100 target cells per minute can be sorted out from the sample solution and thus is particularly suitable for sorting very rare target cells, such as circulating tumor cells. The actuation of the electromagnetic valve has no influence on RPS cell detection and the consequent cell-sorting process. The viability of the collected A549 cell is not impacted by the applied electric field when the cell passes the RPS detection area. The device described in this article is simple, automatic, and label-free and has wide applications in size-based rare target cell sorting for medical diagnostics.

  17. Dynamic in-situ sensing of fluid-dispersed 2D materials integrated on microfluidic Si chip

    PubMed Central

    Hogan, Benjamin T.; Dyakov, Sergey A.; Brennan, Lorcan J.; Younesy, Salma; Perova, Tatiana S.; Gun’ko, Yurii K.; Craciun, Monica F.; Baldycheva, Anna

    2017-01-01

    In this work, we propose a novel approach for wafer-scale integration of 2D materials on CMOS photonic chip utilising methods of synthetic chemistry and microfluidics technology. We have successfully demonstrated that this approach can be used for integration of any fluid-dispersed 2D nano-objects on silicon-on-insulator photonics platform. We demonstrate for the first time that the design of an optofluidic waveguide system can be optimised to enable simultaneous in-situ Raman spectroscopy monitoring of 2D dispersed flakes during the device operation. Moreover, for the first time, we have successfully demonstrated the possibility of label-free 2D flake detection via selective enhancement of the Stokes Raman signal at specific wavelengths. We discovered an ultra-high signal sensitivity to the xyz alignment of 2D flakes within the optofluidic waveguide. This in turn enables precise in-situ alignment detection, for the first practicable realisation of 3D photonic microstructure shaping based on 2D-fluid composites and CMOS photonics platform, while also representing a useful technological tool for the control of liquid phase deposition of 2D materials. PMID:28186118

  18. Continuous-Flow Synthesis of N-Succinimidyl 4-[18F]fluorobenzoate Using a Single Microfluidic Chip

    PubMed Central

    Kimura, Hiroyuki; Tomatsu, Kenji; Saiki, Hidekazu; Arimitsu, Kenji; Ono, Masahiro; Kawashima, Hidekazu; Iwata, Ren; Nakanishi, Hiroaki; Ozeki, Eiichi; Kuge, Yuji; Saji, Hideo

    2016-01-01

    In the field of positron emission tomography (PET) radiochemistry, compact microreactors provide reliable and reproducible synthesis methods that reduce the use of expensive precursors for radiolabeling and make effective use of the limited space in a hot cell. To develop more compact microreactors for radiosynthesis of 18F-labeled compounds required for the multistep procedure, we attempted radiosynthesis of N-succinimidyl 4-[18F]fluorobenzoate ([18F]SFB) via a three-step procedure using a microreactor. We examined individual steps for [18F]SFB using a batch reactor and microreactor and developed a new continuous-flow synthetic method with a single microfluidic chip to achieve rapid and efficient radiosynthesis of [18F]SFB. In the synthesis of [18F]SFB using this continuous-flow method, the three-step reaction was successfully completed within 6.5 min and the radiochemical yield was 64 ± 2% (n = 5). In addition, it was shown that the quality of [18F]SFB synthesized on this method was equal to that synthesized by conventional methods using a batch reactor in the radiolabeling of bovine serum albumin with [18F]SFB. PMID:27410684

  19. Dynamic in-situ sensing of fluid-dispersed 2D materials integrated on microfluidic Si chip

    NASA Astrophysics Data System (ADS)

    Hogan, Benjamin T.; Dyakov, Sergey A.; Brennan, Lorcan J.; Younesy, Salma; Perova, Tatiana S.; Gun’Ko, Yurii K.; Craciun, Monica F.; Baldycheva, Anna

    2017-02-01

    In this work, we propose a novel approach for wafer-scale integration of 2D materials on CMOS photonic chip utilising methods of synthetic chemistry and microfluidics technology. We have successfully demonstrated that this approach can be used for integration of any fluid-dispersed 2D nano-objects on silicon-on-insulator photonics platform. We demonstrate for the first time that the design of an optofluidic waveguide system can be optimised to enable simultaneous in-situ Raman spectroscopy monitoring of 2D dispersed flakes during the device operation. Moreover, for the first time, we have successfully demonstrated the possibility of label-free 2D flake detection via selective enhancement of the Stokes Raman signal at specific wavelengths. We discovered an ultra-high signal sensitivity to the xyz alignment of 2D flakes within the optofluidic waveguide. This in turn enables precise in-situ alignment detection, for the first practicable realisation of 3D photonic microstructure shaping based on 2D-fluid composites and CMOS photonics platform, while also representing a useful technological tool for the control of liquid phase deposition of 2D materials.

  20. Dynamic in-situ sensing of fluid-dispersed 2D materials integrated on microfluidic Si chip.

    PubMed

    Hogan, Benjamin T; Dyakov, Sergey A; Brennan, Lorcan J; Younesy, Salma; Perova, Tatiana S; Gun'ko, Yurii K; Craciun, Monica F; Baldycheva, Anna

    2017-02-10

    In this work, we propose a novel approach for wafer-scale integration of 2D materials on CMOS photonic chip utilising methods of synthetic chemistry and microfluidics technology. We have successfully demonstrated that this approach can be used for integration of any fluid-dispersed 2D nano-objects on silicon-on-insulator photonics platform. We demonstrate for the first time that the design of an optofluidic waveguide system can be optimised to enable simultaneous in-situ Raman spectroscopy monitoring of 2D dispersed flakes during the device operation. Moreover, for the first time, we have successfully demonstrated the possibility of label-free 2D flake detection via selective enhancement of the Stokes Raman signal at specific wavelengths. We discovered an ultra-high signal sensitivity to the xyz alignment of 2D flakes within the optofluidic waveguide. This in turn enables precise in-situ alignment detection, for the first practicable realisation of 3D photonic microstructure shaping based on 2D-fluid composites and CMOS photonics platform, while also representing a useful technological tool for the control of liquid phase deposition of 2D materials.

  1. Fast Screening Techniques for Neurotoxigenic Substances and Other Toxicants and Pollutants Based on Thermal Lensing and Microfluidic Chips.

    PubMed

    Franko, Mladen; Liu, Mingqiang; Boškin, Aleš; Delneri, Ambra; Proskurnin, Mikhail A

    2016-01-01

    Efficient environment protection and human safety require high-throughput analysis techniques for pollutants or toxicants for large sample sets. State-of-the-art HPLC and GC coupled to various detecting strategies offer excellent sensitivity and selectivity, though they are quite time-extensive (2 - 3 samples/h or less when sample preparation is involved). Efforts are made towards screening techniques with high sample throughputs simultaneously providing detection limits below the maximum contaminant levels for the analyte. However, such approaches frequently sacrifice the selectivity or sensitivity (or just give a yes/no response). In this review, we demonstrate thermal-lens spectrometry and microscopy as highly sensitive spectrometric techniques in combination with flow-injection analysis (FIA) and microfluidic FIA along with lab-on-a-chip chemistry for fast screening (several samples/h and up to 20 samples/min) exemplified by organophosphates and carbamates as neurotoxigenic compounds. Various approaches to determining other topical toxicants, like microcystin and cyanopigments as its indicators, allergens, and carcinogenic chromate, are also discussed.

  2. Integration of microfluidic chip with biomimetic hydrogel for 3D controlling and monitoring of cell alignment and migration.

    PubMed

    Lee, Kwang Ho; Lee, Ki Hwa; Lee, Jeonghoon; Choi, Hyuk; Lee, Donghee; Park, Yongdoo; Lee, Sang-Hoon

    2014-04-01

    A biomimetic hydrogel was integrated into microfluidic chips to monitor glioma cell alignment and migration. The extracellular matrix-based biomimetic hydrogel was remodeled by matrix metalloprotease (MMP) secreted by glioma cells and the hydrogel could thus be used to assess cellular behavior. Both static and dynamic cell growth conditions (flow rate of 0.1 mL/h) were used. Cell culture medium with and without vascular endothelial growth factor (VEGF), insensitive VEGF and tissue inhibitor of metalloproteinases (TIMP) were employed to monitor cell behavior. A concentration gradient formed in the hydrogel resulted in differences in cell behavior. Glioma cell viability in the microchannel was 75-85%. Cells in the VEGF-loaded microchannels spread extensively, degrading the MMP-sensitive hydrogel, and achieved cell sizes almost fivefold larger than seen in the control medium. Our integrated system can be used as a model for the study of cellular behavior in a controlled microenvironment generated by fluidic conditions in a biomimetic matrix.

  3. Development of flow through dielectrophoresis microfluidic chips for biofuel production: Sorting and detection of microalgae with different lipid contents

    PubMed Central

    Deng, Yu-Luen; Kuo, Mei-Yi; Juang, Yi-Je

    2014-01-01

    In this study, a continuous flow dielectrophoresis (DEP) microfluidic chip was fabricated and utilized to sort out the microalgae (C. vulgaris) with different lipid contents. The proposed separation scheme is to allow that the microalgae with different lipid contents experience different negative or no DEP force at the separation electrode pair under the pressure-driven flow. The microalgae that experience stronger negative DEP will be directed to the side channel while those experience less negative or no DEP force will pass through the separation electrode pair to remain in the main channel. It was found that the higher the lipid content inside the microalgae, the higher the crossover frequency. Separation of the microalgae with 13% and 21% lipid contents, and 24% and 30%–35% lipid contents was achieved at the operating frequency 7 MHz, and 10 MHz, respectively. Moreover, separation can be further verified by measurement of the fluorescence intensity of the neutral lipid inside the sorted algal cells. PMID:25553195

  4. Tree-on-a-chip: a microfluidic osmotic pump mimicking passive phloem loaders

    NASA Astrophysics Data System (ADS)

    Comtet, Jean; Jensen, Kaare H.; Stroock, Abraham D.; Hosoi, Anette (Peko)

    2014-11-01

    According to the Münch mechanism, vascular plants rely on osmotic pressure gradients to export sugars from regions of synthesis (mature leaves) to regions of consumption (roots, fruits). A crucial step in this process is the loading of sugars from photosynthetic cells (known as mesophylls) to the export conduit (the phloem). In some plants, known as passive loaders, sugars are thought to simply diffuse from mesophylls to the phloem. In this case, we show that a single nondimensional ``flushing number,'' characterizing the relative balance of diffusive sugar loading and convective phloem transport, accurately describes the state of the system (phloem hydrostatic pressure, sugar export rates...). We build a synthetic microfluidic osmotic pump mimicking this biological transport mechanism. In particular, our pump can work in a diffusion-limited regime, for which the flow rate scales weakly with the resistance of the hydraulic circuit. This bio-inspired device provides insight into the biophysical mechanism of passive phloem loading, and could be relevant for microfluidic or micro-robotic applications, where high actuation pressures and steady-state flow rates are necessary.

  5. Manipulation of liquid-liquid interfaces for tunable optics on a chip in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Tan, Kam Yan Sindy

    This thesis describes the design and development of optofluidics: a new class of optical components based on dynamic liquid-liquid interfaces between liquids possessing different optical properties in microfluidic systems. Devices with optical interfaces formed by liquids possess characteristics that are quite different from solid-gas and solid-liquid systems commonly used in conventional optics. Advantages of optofluidic systems include the simplicity to reconfigure optical properties and functions in real time. Examples of devices include liquid waveguides, lenses, and multi-color droplet dye laser. Potential applications include biochemical characterization and optical spectroscopy in micro-total analytical systems. Chapter 1 describers the motivation, general configuration and characteristics of devices with optical interfaces formed by liquids in microchannels. Chapter 2 describes the soft lithographic techniques used to make optofluidic devices. Chapter 3 describes the use of co-fabrication to design and fabricate multiple functional components in microfluidic systems in a single step. Chapters 4 to 6 describe the design and operation of three optofluidic devices: waveguides, lenses, and dye lasers.

  6. General concept of high-performance amperometric detector for microfluidic (bio)analytical chips.

    PubMed

    Amatore, Christian; Da Mota, Nicolas; Sella, Catherine; Thouin, Laurent

    2008-07-01

    In this work, we established theoretically that amperometric detector arrays consisting of a series of parallel band microelectrodes placed on the wall of a microchannel may offer excellent analytical detection performances when implemented onto microfluidic (bio)analytical devices after the separative stages. In combination with the concentration imprinting strategies reported in a previous work, these exceptional performances may be extended to nonelectroactive or poorly diffusing analytes. Using an array of electrodes instead of a large single band allows the whole core of the channel to be probed though keeping an excellent time resolution. Thus, analytes with close retention times may be characterized individually with a resolution which eventually outpaces that of spectroscopic detections. Such important advantages may be obtained only through a complete understanding of the complex coupling between diffusional and convective transport of molecules in microfluidic solutions near an electrochemical detector. As a consequence, the conditions underlying the theoretical data presented in this work have been selected after optimizing procedures rooted on previous theoretical analyses. They will be fully disclosed in a series of further works that will also establish the experimental performances of such amperometric detectors and validate the present concept.

  7. Hand-powered microfluidics: A membrane pump with a patient-to-chip syringe interface

    NASA Astrophysics Data System (ADS)

    MacDonald, Brendan; Gong, Max; Nguyen, Trung; Sinton, David

    2012-11-01

    In this talk, an on-chip hand-powered membrane pump with a robust patient-to-chip syringe interface is presented. This approach enables safe sample collection, sample containment, integrated sharps disposal, high sample volume capacity, and controlled downstream flow with no electrical power requirements. Sample is manually injected into the device via a syringe and needle. The membrane pump inflates upon injection and subsequently deflates, delivering fluid to downstream components in a controlled manner. The device is fabricated from poly(methyl methacrylate) (PMMA) and silicone, using CO2 laser micromachining. Pump performance is experimentally demonstrated and the behavior is subsequently modeled with reference to a resistor-capacitor electrical circuit analogy. Downstream output of the membrane pump is regulated, and scaled, by connecting multiple pumps in parallel. The device provides precisely controlled pumping rates and high volume throughput without any electrical power requirements.

  8. Characterization and evaluation of two-dimensional microfluidic chip-HPLC coupled to tandem mass spectrometry for quantitative analysis of 7-aminoflunitrazepam in human urine.

    PubMed

    Bai, Hsin-Yu; Lin, Shu-Ling; Chan, Shen-An; Fuh, Ming-Ren

    2010-10-01

    Microfluidic chip-based high-performance-liquid-chromatography coupled to mass spectrometry (chip-HPLC-MS) has been widely used in proteomic research due to its enhanced sensitivity. We employed a chip-HPLC-MS system for determining small molecules such as drug metabolites in biological fluids. This chip-HPLC-MS system integrates a microfluidic switch, a 2-dimensional column design including an enrichment column (160 nL) for sample pre-concentration and an analytical column for chromatographic separation, as well as a nanospray emitter on a single polyimide chip. In this study, a relatively large sample volume (500 nL) was injected into the enrichment column for pre-concentration and an additional 4 μL of the initial mobile phase was applied to remove un-retained components from the sample matrix prior to chromatographic separation. The 2-dimensional column design provides the advantages of online sample concentration and reducing matrix influence on MS detection. 7-Aminoflunitrazepam (7-aminoFM2), a major metabolite of flunitrazepam (FM2), was determined in urine samples using the integrated chip-HPLC-MS system. The linear range was 0.1-10 ng mL(-1) and the method detection limit (signal-to-noise ratio of 3) was 0.05 ng mL(-1) for 7-aminoFM2. After consecutive liquid-liquid extraction (LLE) and solid-phase extraction (SPE), the chip-HPLC-MS exhibited high correlation between 7-aminoFM2 spiked Milli-Q water and 7-aminoFM2 spiked urine samples. This system also showed good precision (n = 5) and recovery for spiked urine samples at the levels of 0.1, 1.0, and 10 ng mL(-1). Intra-day and inter-day precision were 2.0-7.1% and 4.3-6.0%, respectively. Clinical urine samples were also analyzed by this chip-HPLC-MS system and acceptable relative differences (-1.3 to -13.0%) compared with the results using a GC-MC method were determined. Due to its high sensitivity and ease of operation, the chip-HPLC-MS system can be utilized for the determination of small molecules such

  9. 96-Well Polycarbonate-Based Microfluidic Titer Plate for High-Throughput Purification of DNA and RNA

    PubMed Central

    Witek, Małgorzata A.; Hupert, Mateusz L.; Park, Daniel S.-W.; Fears, Kirby; Murphy, Michael C.; Soper, Steven A.

    2008-01-01

    We report a simple and effective method for the high-throughput purification of a variety of nucleic acids (NAs) from whole cell lysates or whole blood using a photactivated polycarbonate solid-phase reversible immobilization (PPC-SPRI) microfluidic chip. High-throughput operation was achieved by placing 96 purification beds, each containing an array of 3800 20 µm diameter posts, on a single 3″ × 5″ polycarbonate (PC) wafer fabricated by hot embossing. All beds were interconnected through a common microfluidic network that permitted parallel access through the use of a vacuum and syringe pump for delivery of immobilization buffer (IB) and effluent. The PPC-SPRI purification was accomplished by condensation of NAs onto a UV-modified PC surface in the presence of the IB comprised of polyethylene glycol, NaCl, and ethanol with a composition dependent on the length of the NAs to be isolated and the identity of the sample matrix. The performance of the device was validated by quantification of the recovered material following PCR (for DNA) or RT-PCR (for RNA). The extraction bed load capacity of NAs was 206 ± 93 ng for gDNA and 165 ± 81 ng for TRNA from Escherichia coli. Plate-to-plate variability was found to be 35 ± 10%. The purification process was fast (>30 min) and easy to automate, and the low cost of wafer fabrication makes it appropriate for single-use applications. PMID:18355091

  10. Apoptosis goes on a chip: advances in the microfluidic analysis of programmed cell death

    PubMed Central

    Wlodkowic, Donald; Khoshmanesh, Khashayar; Sharpe, John C; Darzynkiewicz, Zbigniew; Cooper, Jonathan Mark

    2011-01-01

    Summary Recent years have brought enormous progress in cell-based lab-on-a-chip technologies, allowing dynamic studies of cell death with an unprecedented accuracy. As interest in the microfabricated technologies for cell-based bioassays is rapidly gaining momentum, we highlight the most promising technologies that provide a new outlook for the rapid assessment of programmed and accidental cell death and are applicable in drug discovery, high-content drug screening, and personalized clinical diagnostics. PMID:21630641

  11. Hand-powered microfluidics: A membrane pump with a patient-to-chip syringe interface

    PubMed Central

    Gong, Max M.; MacDonald, Brendan D.; Vu Nguyen, Trung; Sinton, David

    2012-01-01

    In this paper, we present an on-chip hand-powered membrane pump using a robust patient-to-chip syringe interface. This approach enables safe sample collection, sample containment, integrated sharps disposal, high sample volume capacity, and controlled downstream flow with no electrical power requirements. Sample is manually injected into the device via a syringe and needle. The membrane pump inflates upon injection and subsequently deflates, delivering fluid to downstream components in a controlled manner. The device is fabricated from poly(methyl methacrylate) (PMMA) and silicone, using CO2 laser micromachining, with a total material cost of ∼0.20 USD/device. We experimentally demonstrate pump performance for both deionized (DI) water and undiluted, anticoagulated mouse whole blood, and characterize the behavior with reference to a resistor-capacitor electrical circuit analogy. Downstream output of the membrane pump is regulated, and scaled, by connecting multiple pumps in parallel. In contrast to existing on-chip pumping mechanisms that typically have low volume capacity (∼5 μL) and sample volume throughput (∼1–10 μl/min), the membrane pump offers high volume capacity (up to 240 μl) and sample volume throughput (up to 125 μl/min). PMID:24143160

  12. Comparative toxicity of lead (Pb(2+)), copper (Cu(2+)), and mixtures of lead and copper to zebrafish embryos on a microfluidic chip.

    PubMed

    Li, Yinbao; Yang, Xiujuan; Chen, Zuanguang; Zhang, Beibei; Pan, Jianbin; Li, Xinchun; Yang, Fan; Sun, Duanping

    2015-03-01

    Investigations were conducted to determine acute effects of Pb(2+) and Cu(2+) presented individually and collectively on zebrafish embryos. Aquatic safety testing requires a cheap, fast, and highly efficient platform for real-time evaluation of single and mixture of metal toxicity. In this study, we have developed a microfluidic system for phenotype-based evaluation of toxic effects of Pb(2+) and Cu(2+) using zebrafish (Danio rerio) embryos. The microfluidic chip is composed of a disc-shaped concentration gradient generator and 24 culture chambers, which can generate one blank solution, seven mixture concentrations, and eight single concentrations for each metal solution, thus enabling the assessment of zebrafish embryos. To test the accuracy of this new chip platform, we have examined the toxicity and teratogenicity of Pb(2+) and Cu(2+) on embryos. The individual and combined impact of Pb(2+) and Cu(2+) on zebrafish embryonic development was quantitatively assessed by recording a series of physiological indicators, such as spontaneous motion at 22 hours post fertilization (hpf), mortality at 24 hpf, heartbeat and body length at 96 hpf, etc. It was found that Pb(2+) or Cu(2+) could induce deformity and cardiovascular toxicity in zebrafish embryos and the mixture could induce more severe toxicity. This chip is a multiplexed testing apparatus that allows for the examination of toxicity and teratogenicity for substances and it also can be used as a potentially cost-effective and rapid aquatic safety assessment tool.

  13. Measurement of temperature-dependent diffusion coefficients using a confocal Raman microscope with microfluidic chips considering laser-induced heating effect.

    PubMed

    Lin, Ying; Yu, Xinhai; Wang, Zhenyu; Tu, Shan-Tung; Wang, Zhengdong

    2010-05-14

    Conventional methods for measuring diffusion coefficients (D) are complex and time consuming. This study presents a method for the continuous measurement of temperature-dependent diffusion coefficients using a confocal Raman microscope with microfluidic chips. Concentration information was collected by a Raman microscope to extract D values. An isothermal diffusion process at various temperatures was ensured by coupling the silicon-based microfluidic chip with an isothermal plate. In the simple silicon/glass chip, the heating effect induced by a Raman laser was observed to contribute to abnormally high D values. To eliminate the heating effect, a 200nm-thick aluminum (Al) reflection film was used to coat the channel bottom. The Al film substantially reduced absorption of laser power, thus ensuring precise D values in excellent agreement with literature data. Other potential methods to eliminate the heating effect were also evaluated by computational fluid dynamics (CFD) simulations and were found impractical for implementation. Consequently, this method for the continuous measurement of temperature-dependent diffusion coefficients is proven to be accurate, efficient, and reliable.

  14. Dielectrophoresis microjets: a merging of electromagnetics and microfluidics for on-chip technologies

    NASA Astrophysics Data System (ADS)

    Hill, Kyle A.; Collier, Christopher M.; Holzman, Jonathan F.

    2014-05-01

    Digital (droplet-based) microfluidic systems apply electromagnetic characteristics as the fundamental fluid actuation mechanism. These systems are often implemented in two-dimensional architectures, overcoming one-dimensional continuous flow channel practical issues. The fundamental operation for digital microfluidics requires the creation of an electric field distribution to achieve desired fluid actuation. The electric field distribution is typically non-uniform, enabling creation of net dielectrophoresis (DEP) force. The DEP force magnitude is proportional to the difference between microdroplet and surrounding medium complex dielectric constants, and the gradient of the electric field magnitude squared. Force sign/direction can be manipulated to achieve a force towards higher (positive DEP) or lower (negative DEP) electrostatic energy by tailoring the relative difference between microdroplet and surrounding medium complex dielectric constants through careful selection of the devices fabrication materials. The DEP force magnitudes and directions are applied here for well-controlled and high-speed microdroplet actuation. Control and speed characteristics arise from significant differences in the microdroplet/medium conductivity and the use of a micropin architecture with strong electric field gradients. The implementation, referred to here as a DEP microjet, establishes especially strong axial propulsion forces. Single- and double-micropin topologies achieve strong axial propulsion force, but only the double-micropin topology creates transverse converging forces for stable and controlled microdroplet actuation. Electric field distributions for each topology are investigated and linked to axial and transverse forces. Experimental results are presented for both topologies. The double-micropin topology is tested with biological fluids. Microdroplet actuation speeds up to 25 cm/s are achieved—comparable to the fastest speeds to-date.

  15. Flame Aerosol Deposition of TiO2 Nanoparticle Films on Polymers and Polymeric Microfluidic Devices for On-Chip Phosphopeptide Enrichment.

    PubMed

    Rudin, Thomas; Tsougeni, Katerina; Gogolides, Evangelos; Pratsinis, Sotiris E

    2012-09-01

    Direct and fast (10s of seconds) deposition of flame-made, high surface-area aerosol films on polymers and polymeric microfluidic devices is demonstrated. Uniform TiO2 nanoparticle films were deposited on cooled Poly(methyl methacrylate) (PMMA) substrates by combustion of titanium(IV) isopropoxide (TTIP) - xylene solution sprays. Films were mechanically stabilized by in-situ annealing with a xylene spray flame. Plasma-etched microfluidic chromatography columns, comprising parallel microchannels were also coated with such nanoparticle films without any microchannel deformation. These microcolumns were successfully used in metal-oxide affinity chromatography (MOAC) to selectively trap phosphopeptides on these high surface-area nanostructured films. The chips had a high capacity retaining 1.2 μg of standard phosphopeptide. A new extremely fast method is developed for MOAC microchip stationary phase fabrication with applications in proteomics.

  16. Flame Aerosol Deposition of TiO2 Nanoparticle Films on Polymers and Polymeric Microfluidic Devices for On-Chip Phosphopeptide Enrichment

    PubMed Central

    Rudin, Thomas; Tsougeni, Katerina; Gogolides, Evangelos; Pratsinis, Sotiris E.

    2013-01-01

    Direct and fast (10s of seconds) deposition of flame-made, high surface-area aerosol films on polymers and polymeric microfluidic devices is demonstrated. Uniform TiO2 nanoparticle films were deposited on cooled Poly(methyl methacrylate) (PMMA) substrates by combustion of titanium(IV) isopropoxide (TTIP) – xylene solution sprays. Films were mechanically stabilized by in-situ annealing with a xylene spray flame. Plasma-etched microfluidic chromatography columns, comprising parallel microchannels were also coated with such nanoparticle films without any microchannel deformation. These microcolumns were successfully used in metal-oxide affinity chromatography (MOAC) to selectively trap phosphopeptides on these high surface-area nanostructured films. The chips had a high capacity retaining 1.2 μg of standard phosphopeptide. A new extremely fast method is developed for MOAC microchip stationary phase fabrication with applications in proteomics. PMID:23729946

  17. Solid-state sensor incorporated in microfluidic chip and magnetic-bead enzyme immobilization approach for creatinine and glucose detection in serum

    NASA Astrophysics Data System (ADS)

    Lin, Yen-Heng; Chiang, Chien-Hung; Wu, Min-Hsien; Pan, Tung-Ming; Luo, Ji-Dung; Chiou, Chiuan-Chian

    2011-12-01

    Solid-state sensors are stable and inexpensive electric transducers for biomedical measurement. This study proposes a microfluidic chip incorporated with a solid-state sensor for measuring glucose and creatinine in blood serum. Magnetic beads are employed to immobilize enzymes and deliver them in a micro-channel. Glucose and creatinine can be measured at 2-8 mM and 10-2 to 10 mM, respectively, which is a meaningful range in human blood. The immobilization approach also addresses the issue of the long-term preservation of enzymes in microfluidic devices. The proposed device is suitable for multi-target measurement in a point-of-care system.

  18. Quantitative determination of 8-isoprostaglandin F(2α) in human urine using microfluidic chip-based nano-liquid chromatography with on-chip sample enrichment and tandem mass spectrometry.

    PubMed

    Bai, Hsin-Yu; Lin, Shu-Ling; Chung, Yu-Ting; Liu, Tsung-Yun; Chan, Shan-An; Fuh, Ming-Ren

    2011-04-15

    Urinary 8-isoprostaglandin F(2α) (8-isoPGF(2α)) has been reported as an important biomarker to indicate the oxidative stress status in vivo. In order to quantitatively determine the low contents of 8-isoPGF(2α) (in sub- to low ng mL(-1) range) in physiological fluids, a sensitive detection method has become an important issue. In this study, we employed a microfluidic chip-based nano liquid chromatography (chip-nanoLC) with on-chip sample enrichment coupled to triple quadrupole mass spectrometer (QqQ-MS) for the quantitative determination of 8-isoPGF(2α) in human urine. This chip-nanoLC unit integrates a microfluidic switch, a chip column design having a pre-column (enrichment column) for sample enrichment prior to an analytical column for separation, as well as a nanospray emitter on a single polyimide chip. The introduction of enrichment column offers the advantages of online sample pre-concentration and reducing matrix influence on MS detection to improve sensitivity. In this study, the chip-nanoLC consisting of Zorbax 300A SB-C18 columns and Agilent QqQ Mass spectrometer were used for determining 8-isoPGF(2α) in human urine. Gradient elution was employed for effective LC separation and multiple reaction monitoring (MRM) was utilized for the quantitative determination of 8-isoPGF(2α) (m/z 353→193). We employed liquid-liquid extraction (LLE)/solid-phase extraction (SPE) for extracting analyte and reducing matrix effect from urine sample prior to chip-nanoLC/QqQ-MS analysis for determining urinary 8-isoPGF(2α). Good recoveries were found to be in the range of 83.0-85.3%. The linear range was 0.01-2 ng mL(-1) for urinary 8-isoPGF(2α). In addition, the proposed method showed good precision and accuracy for 8-isoPGF(2α) spiked synthetic urine samples. Intra-day and inter-day precisions were 1.8-5.0% and 4.3-5.8%, respectively. The method accuracy for intra-day and inter-day assays ranged from 99.3 to 99.9% and 99.4 to 99.7%, respectively. Due to its

  19. A microfluidic co-culture system to monitor tumor-stromal interactions on a chip

    PubMed Central

    Menon, Nishanth V.; Cao, Bin; Lim, Mayasari; Kang, Yuejun

    2014-01-01

    The living cells are arranged in a complex natural environment wherein they interact with extracellular matrix and other neighboring cells. Cell-cell interactions, especially those between distinct phenotypes, have attracted particular interest due to the significant physiological relevance they can reveal for both fundamental and applied biomedical research. To study cell-cell interactions, it is necessary to develop co-culture systems, where different cell types can be cultured within the same confined space. Although the current advancement in lab-on-a-chip technology has allowed the creation of in vitro models to mimic the complexity of in vivo environment, it is still rather challenging to create such co-culture systems for easy control of different colonies of cells. In this paper, we have demonstrated a straightforward method for the development of an on-chip co-culture system. It involves a series of steps to selectively change the surface property for discriminative cell seeding and to induce cellular interaction in a co-culture region. Bone marrow stromal cells (HS5) and a liver tumor cell line (HuH7) have been used to demonstrate this co-culture model. The cell migration and cellular interaction have been analyzed using microscopy and biochemical assays. This co-culture system could be used as a disease model to obtain biological insight of pathological progression, as well as a tool to evaluate the efficacy of different drugs for pharmaceutical studies. PMID:25553194

  20. Development-on-chip: in vitro neural tube patterning with a microfluidic device

    PubMed Central

    Soundararajan, Prabakaran; Chennampally, Phaneendra; Cox, Gregory A.

    2016-01-01

    Embryogenesis is a highly regulated process in which the precise spatial and temporal release of soluble cues directs differentiation of multipotent stem cells into discrete populations of specialized adult cell types. In the spinal cord, neural progenitor cells are directed to differentiate into adult neurons through the action of mediators released from nearby organizing centers, such as the floor plate and paraxial mesoderm. These signals combine to create spatiotemporal diffusional landscapes that precisely regulate the development of the central nervous system (CNS). Currently, in vivo and ex vivo studies of these signaling factors present some inherent ambiguity. In vitro methods are preferred for their enhanced experimental clarity but often lack the technical sophistication required for biological realism. In this article, we present a versatile microfluidic platform capable of mimicking the spatial and temporal chemical environments found in vivo during neural tube development. Simultaneous opposing and/or orthogonal gradients of developmental morphogens can be maintained, resulting in neural tube patterning analogous to that observed in vivo. PMID:27246712

  1. Numerical Simulation on the Response Characteristics of a Pneumatic Microactuator for Microfluidic Chips.

    PubMed

    Liu, Xuling; Li, Songjing; Bao, Gang

    2016-06-01

    This article presents a multiphysical system modeling and simulation of a pneumatic microactuator, which significantly influences the performance of a particular pneumatic microfluidic device. First, the multiphysical system modeling is performed by developing a physical model for each of its three integrated components: microchannel with a microvalve, a gas chamber, and an elastomer membrane. This is done for each step of operation for the whole system. The whole system is then considered a throttle blind capacitor model, and it is used to predict the response time of the pneumatic microactuator by correlating its characteristics such as gas pressurizing, hydraulic resistance, and membrane deformation. For this microactuator, when the maximum membrane deformation is 100 µm, the required actuated air pressure is 80 kPa, and the response time is 1.67 ms when the valve-opening degree is 0.8. The response time is 1.61 ms under fully open conditions. These simulated results are validated by the experimental results of the current and previous work. A correlation between the simulated and experimental results confirms that the multiphysical modeling presented in this work is applicable in developing a proper design of a pneumatic microactuator. Finally, the influencing factors of the response time are discussed and analyzed.

  2. A microfabricated microfluidic bioMEMS device to model human brain aneurisms: the aneurysm-on-a-chip

    NASA Astrophysics Data System (ADS)

    Reece, Lisa M.; Khor, Jian Wei; Thakur, Raviraj; Amin, Ahmed; Wereley, Steven T.; Leary, James F.

    2015-03-01

    Aneurysms are pockets of blood that collect outside blood vessel walls forming dilatations and leaving arterial walls very prone to rupture. There is little information concerning the causes of intracranial aneurysm formation, growth, and rupture. Current treatments include: (1) clipping, and (2) coil embolization, including stent-assisted coiling. Further, the evolution of any aneurysm is assumed to be caused by the remodeling of the affected blood vessel's material constituents (tunica intima, tunica media, or tunica adventitia). Velocity, pressure, and wall shear stresses aid in the disease development of aneurysmal growth, while the shear force mechanisms effecting wound closure are elusive. To study aneurysm pathogenesis, a lab-on-a-chip device is the key to discovering the underlying mechanisms of these lesions. A two-dimensional microfluidic model, the Aneurysm-on-a-Chip™ (AOC), was the logical answer to study particle flow within an aneurysm "sac". The AOC apparatus can track particles/cells when it is coupled to particle image velocimetry software (PIV) package. The AOC fluid flow was visualized using standard microscopy techniques with commercial microparticles and human aortic smooth muscle cells (HASMC). Images were taken during fluid flow experiments and PIV was utilized to monitor the flow of particles within the "sac" region, as well as particles entering and exiting the device. Quiver plots were generated from fluid flow experiments using standard 7 μm latex particles and fixed HASMC in PBS. PIV analysis shows that the particles flowed nicely from input to output. Wall shear stress provided evidence that there was some back flow at the edges of the "sac" - an indicator of aneurysm development in human patients.

  3. Logic digital fluidic in miniaturized functional devices: Perspective to the next generation of microfluidic lab-on-chips.

    PubMed

    Zhang, Qiongdi; Zhang, Ming; Djeghlaf, Lyas; Bataille, Jeanne; Gamby, Jean; Haghiri-Gosnet, Anne-Marie; Pallandre, Antoine

    2017-04-01

    Microfluidics has emerged following the quest for scale reduction inherent to micro- and nanotechnologies. By definition, microfluidics manipulates fluids in small channels with dimensions of tens to hundreds of micrometers. Recently, microfluidics has been greatly developed and its influence extends not only the domains of chemical synthesis, bioanalysis, and medical researches but also optics and information technology. In this review article, we will shortly discuss an enlightening analogy between electrons transport in electronics and fluids transport in microfluidic channels. This analogy helps to master transport and sorting. We will present some complex microfluidic devices showing that the analogy is going a long way off toward more complex components with impressive similarities between electronics and microfluidics. We will in particular explore the vast manifold of fluidic operations with passive and active fluidic components, respectively, as well as the associated mechanisms and corresponding applications. Finally, some relevant applications and an outlook will be cited and presented.

  4. Selective filling for patterning in microfluidic channels and integration of chromatography in "lab-on-a-chip" devices using sol-gel technology

    NASA Astrophysics Data System (ADS)

    Jindal, Rohit

    The last decade has seen tremendous advancement in the development of miniaturized chemical analysis system also known as "lab-on-a-chip". It is believed that the true potential of these devices will be achieved by integrating various functions such as separation, reaction, sensing, mixing, pumping, injection and detection onto a single chip. The ability to pattern different functionalities is indispensable for the development of highly integrated devices. In this work, a simple method based on the concept of selective filling is described for patterning in the microfluidic channels. It is based on the difference in the free energy of filling between an open and a covered part of the channel. This method was used for the integration of chromatography in the microfluidic devices. A chromatographic column was realized by utilizing sol-gel as an immobilization matrix for entrapping reversed phase chromatographic particles. Localization of the stationary phase was achieved using the selective filling technique. Channels were fabricated in quartz using photolithography and wet etching. Electroosmotic flow was used for manipulating fluid movement in the channels. Cross channel design was used for making a pulse injection of the solutes in the separation channel. An optical fiber setup was developed for carrying out on-chip UV absorbance detection. Stationary phase was created under different sol-gel synthesis conditions. It was established that the sol-gel synthesis carried out under acidic conditions provides the optimum synthesis conditions for creating separation column. Chromatographic performance of the stationary phase material was demonstrated by separating peptides present in a mixture. The sol-gel immobilization method was extended for the integration of micropump in the chip. The micropump enables pumping of the fluid in field free channels. Preliminary results, demonstrating the potential of carbon nanotubes as a support material in the microfluidic channels

  5. Strategy for allosteric analysis based on protein-patterned stationary phase in microfluidic chip.

    PubMed

    Bi, Hongyan; Weng, Xuexiang; Qu, Haiyun; Kong, Jilie; Yang, Pengyuan; Liu, Baohong

    2005-01-01

    An effective method is presented for the on-chip analysis of chiral interactions with a successful depression of nonspecific adsorption. The alumina gel-derived protein network on poly(methyl methacrylate) (PMMA) microchannel was explored to form a protein-stationary phase and then used to carry out electrophoresis for fast enantioseparation coupled with electrochemical detection. On the basis of the chemical modification of a synthesized copolymer containing silane-functionalized scaffold, alumina sol-gel could react readily with the silane groups and form steady microstructure on the chip surface achieving the encapsulation of functional biomolecules. Compared with the native PMMA microchannels, the modified surfaces exhibited much better wettability, more stable and enhanced electroosmotic mobility, and less nonspecific adsorption. The water contact angle and EOF of alumina-gel-derived PMMA substrate were 22 degrees and 4.3 x 10(-4) cm(2) V(-1) s(-1), compared to those of 73 degrees and 1.9 x 10(-4) cm(2) V(-1) s(-1) from the untreated one, respectively. Bovine serum albumin, acting as a target protein, could be stably and homogeneously immobilized in the modified PMMA microchannel to fabricate a protein-stationary phase. Under a mild condition, D- and L-tryptophan were efficiently separated with a resolution of 1.57. The as-prepared microchip can perform chiral separations within short time, indicating that the general protocol has the potential to provide a platform for high throughput screening of enantiomer candidates such as those biochemical drugs with protein targets and the research of receptor interactions.

  6. A microfluidic multiwell chip for enzyme-free detection of mRNA from few cells.

    PubMed

    Haider, Michaela; Ji, Bozhi; Haselgrübler, Thomas; Sonnleitner, Alois; Aberger, Fritz; Hesse, Jan

    2016-12-15

    Isogenic cell populations possess heterogeneous gene expression patterns. Most methods for mRNA expression analysis start with the reverse transcription of mRNA into cDNA, a process that can introduce strong signal variations not related to the actual mRNA levels. Miniaturized lab-on-a-chip systems offer properties - e.g. low sample dilution, low contamination - that enable new reaction schemes for molecular analyses. To enable transcription-free mRNA expression analysis of few single cells, a one-step cell lysis, target labelling and hybridisation approach as well as a corresponding passive multiwell chip with a volume of 25.5 nL/well were developed. The method enabled the parallel analysis of up to 96 samples and 6 target genes per sample. Preceding light microscopy of the living cells allowed correlating mRNA levels and cell number. As a proof-of-principle, the pancreatic cancer cell line Panc-1 was investigated for expression heterogeneity of a reference gene plus 5 genes reported to be overexpressed in cancer stem cells (CSCs). A good correlation (r(51)=0.739, p<0.001; rs(51)=0.744, p<0.001) between the cell number per well and the number of detected reference gene mRNA confirmed the proper function of the device. Moreover, a heterogeneous expression of the CSC-associated target genes was found which matched well with reports on the presence of CSCs in the Panc-1 cell line.

  7. Chemical property based sequence characterization of PpcA and its homolog proteins PpcB-E: A mathematical approach

    PubMed Central

    Pal Choudhury, Pabitra

    2017-01-01

    Periplasmic c7 type cytochrome A (PpcA) protein is determined in Geobacter sulfurreducens along with its other four homologs (PpcB-E). From the crystal structure viewpoint the observation emerges that PpcA protein can bind with Deoxycholate (DXCA), while its other homologs do not. But it is yet to be established with certainty the reason behind this from primary protein sequence information. This study is primarily based on primary protein sequence analysis through the chemical basis of embedded amino acids. Firstly, we look for the chemical group specific score of amino acids. Along with this, we have developed a new methodology for the phylogenetic analysis based on chemical group dissimilarities of amino acids. This new methodology is applied to the cytochrome c7 family members and pinpoint how a particular sequence is differing with others. Secondly, we build a graph theoretic model on using amino acid sequences which is also applied to the cytochrome c7 family members and some unique characteristics and their domains are highlighted. Thirdly, we search for unique patterns as subsequences which are common among the group or specific individual member. In all the cases, we are able to show some distinct features of PpcA that emerges PpcA as an outstanding protein compared to its other homologs, resulting towards its binding with deoxycholate. Similarly, some notable features for the structurally dissimilar protein PpcD compared to the other homologs are also brought out. Further, the five members of cytochrome family being homolog proteins, they must have some common significant features which are also enumerated in this study. PMID:28362850

  8. Recreating blood-brain barrier physiology and structure on chip: A novel neurovascular microfluidic bioreactor

    PubMed Central

    Brown, Jacquelyn A.; Pensabene, Virginia; Markov, Dmitry A.; Allwardt, Vanessa; Neely, M. Diana; Shi, Mingjian; Britt, Clayton M.; Hoilett, Orlando S.; Yang, Qing; Brewer, Bryson M.; Samson, Philip C.; McCawley, Lisa J.; May, James M.; Webb, Donna J.; Li, Deyu; Bowman, Aaron B.; Reiserer, Ronald S.; Wikswo, John P.

    2015-01-01

    The blood-brain barrier (BBB) is a critical structure that serves as the gatekeeper between the central nervous system and the rest of the body. It is the responsibility of the BBB to facilitate the entry of required nutrients into the brain and to exclude potentially harmful compounds; however, this complex structure has remained difficult to model faithfully in vitro. Accurate in vitro models are necessary for understanding how the BBB forms and functions, as well as for evaluating drug and toxin penetration across the barrier. Many previous models have failed to support all the cell types involved in the BBB formation and/or lacked the flow-created shear forces needed for mature tight junction formation. To address these issues and to help establish a more faithful in vitro model of the BBB, we have designed and fabricated a microfluidic device that is comprised of both a vascular chamber and a brain chamber separated by a porous membrane. This design allows for cell-to-cell communication between endothelial cells, astrocytes, and pericytes and independent perfusion of both compartments separated by the membrane. This NeuroVascular Unit (NVU) represents approximately one-millionth of the human brain, and hence, has sufficient cell mass to support a breadth of analytical measurements. The NVU has been validated with both fluorescein isothiocyanate (FITC)-dextran diffusion and transendothelial electrical resistance. The NVU has enabled in vitro modeling of the BBB using all human cell types and sampling effluent from both sides of the barrier. PMID:26576206

  9. Polymer-based microfluidic chip for rapid and efficient immunomagnetic capture and release of Listeria monocytogenes.

    PubMed

    Malic, L; Zhang, X; Brassard, D; Clime, L; Daoud, J; Luebbert, C; Barrere, V; Boutin, A; Bidawid, S; Farber, J; Corneau, N; Veres, T

    2015-10-21

    Infections caused by foodborne pathogens such as Listeria monocytogenes pose a threat to public health while timely detection is challenging due to pathogen low numbers. The development of robust and efficient sample preparation techniques is crucial to improve detection sensitivity and workflow. Immunomagnetic separation using magnetic nanoparticles (MNPs) is attractive, as it can efficiently capture target cells. For food safety applications, a platform is needed to rapidly process large sample volumes, allowing capture and release of target bacteria conjugated to immunomagnetic nanoparticles (IMNPs). Herein, we demonstrate a method for magnetic capture and release of bacteria-IMNPs complex based on a 3D magnetic trap integrated on a polymeric microfluidic device. The 3D magnetic capture region consist of a dense array of high-aspect ratio (3 : 1) cylindrical pillars embossed in thermoplastic polymer and coated with soft ferromagnetic nickel by an electroless deposition technique. This allows the generation of strong and switchable magnetic capture regions due to the very low remanence of the nickel shell. We propose and validate an optimized configuration of capture regions for efficient localized capture and rapid release of MNPs and IMNPs conjugated to L. monocytogenes. A maximum recovery rate for MNPs corresponded to 91% while a maximum capture efficiency of 30% was obtained for live bacteria, with a minimum detectable sample concentration of ~10 cfu ml(-1) in 1 ml volume using plate-culture method. We believe that the flexible design and low-cost fabrication process of the proposed system will allow rapid sample preparation for applications beyond food and water safety, including point-of-care diagnosis.

  10. Design, microfabrication, and characterization of a moulded PDMS/SU-8 inkjet dispenser for a Lab-on-a-Printer platform technology with disposable microfluidic chip.

    PubMed

    Bsoul, Anas; Pan, Sheng; Cretu, Edmond; Stoeber, Boris; Walus, Konrad

    2016-08-16

    In this paper, we present a disposable inkjet dispenser platform technology and demonstrate the Lab-on-a-Printer concept, an extension of the ubiquitous Lab-on-a-Chip concept, whereby microfluidic modules are directly integrated into the printhead. The concept is demonstrated here through the integration of an inkjet dispenser and a microfluidic mixer enabling control over droplet composition from a single nozzle in real-time during printing. The inkjet dispenser is based on a modular design platform that enables the low-cost microfluidic component and the more expensive actuation unit to be easily separated, allowing for the optional disposal of the former and reuse of the latter. To limit satellite droplet formation, a hydrophobic-coated and tapered micronozzle was microfabricated and integrated with the fluidics to realize the dispenser. The microfabricated devices generated droplets with diameters ranging from 150-220 μm, depending mainly on the orifice diameter, with printing rates up to 8000 droplets per second. The inkjet dispenser is capable of dispensing materials with a viscosity up to ∼19 mPa s. As a demonstration of the inkjet dispenser function and application, we have printed type I collagen seeded with human liver carcinoma cells (cell line HepG2), to form patterned biological structures.

  11. A Portable Liquid Chromatograph with a Battery-operated Compact Electroosmotic Pump and a Microfluidic Chip Device with a Reversed Phase Packed Column.

    PubMed

    Ishida, Akihiko; Fujii, Mitsutaka; Fujimoto, Takehiro; Sasaki, Shunsuke; Yanagisawa, Ichiro; Tani, Hirofumi; Tokeshi, Manabu

    2015-01-01

    A compact and lightweight liquid chromatography system is presented with overall dimensions of 26 cm width × 18 cm length × 21 cm height and weight of 2 kg. This system comprises a battery-operated compact electroosmotic pump, a manual injector, a microfluidic chip device containing a packed column and an electrochemical detector, and a USB bus-powered potentiostat. The pumping system was designed for microfluidic-based reversed-phase liquid chromatography in which an electroosmotically generated water stream pushes the mobile phase via a diaphragm for the output. The flow rate ranged from 0 to 10 μL/min and had a high degree of precision. The pumping system operated continuously for over 24 h with dry batteries. The column formed in the microfluidic device was packed with 3-μm ODS particles with a length of 30 mm and a diameter of 0.8 mm. The results presented herein demonstrate the performance of the pumping system and the column using alkylphenols, catecholamine, catechin, and amino acids.

  12. Clinical application of a microfluidic chip for immunocapture and quantification of circulating exosomes to assist breast cancer diagnosis and molecular classification

    PubMed Central

    Fang, Shimeng; Tian, Hongzhu; Li, Xiancheng; Jin, Dong; Li, Xiaojie; Kong, Jing; Yang, Chun; Yang, Xuesong; Lu, Yao; Luo, Yong; Lin, Bingcheng; Niu, Weidong

    2017-01-01

    Increasing attention has been attracted by exosomes in blood-based diagnosis because cancer cells release more exosomes in serum than normal cells and these exosomes overexpress a certain number of cancer-related biomarkers. However, capture and biomarker analysis of exosomes for clinical application are technically challenging. In this study, we developed a microfluidic chip for immunocapture and quantification of circulating exosomes from small sample volume and applied this device in clinical study. Circulating EpCAM-positive exosomes were measured in 6 cases breast cancer patients and 3 healthy controls to assist diagnosis. A significant increase in the EpCAM-positive exosome level in these patients was detected, compared to healthy controls. Furthermore, we quantified circulating HER2-positive exosomes in 19 cases of breast cancer patients for molecular classification. We demonstrated that the exosomal HER2 expression levels were almost consistent with that in tumor tissues assessed by immunohistochemical staining. The microfluidic chip might provide a new platform to assist breast cancer diagnosis and molecular classification. PMID:28369094

  13. Sensitive Mie scattering immunoagglutination assay of porcine reproductive and respiratory syndrome virus (PRRSV) from lung tissue samples in a microfluidic chip.

    PubMed

    Song, Jae-Young; Lee, Chang-Hee; Choi, Eun-Jin; Kim, Keesung; Yoon, Jeong-Yeol

    2011-12-01

    A microfluidic immunosensor utilizing Mie scattering immunoaggultination assay was developed for rapid and sensitive detection of porcine reproductive and respiratory syndrome virus (PRRSV) from lung tissue samples of domesticated pigs. Antibodies against PRRSV were conjugated to the surface of highly carboxylated polystyrene microparticles (diameter=920nm) and mixed with the diluted PRRSV tissue samples in a Y-shaped microchannel. Antibody-antigen binding induced microparticle immunoagglutination, which was detected by measuring the forward 45° light scattering of 380nm incident beam using microcallipered, proximity fiber optics. For comparison, multi-well experiments were also performed using the same optical detection setup. The detection limit was determined to be 10(-3)TCID(50)ml(-1) for PRRSV dissolved in PBS, while those of previous RT-PCR studies for PRRSV were 10(1)TCID(50)ml(-1) (conventional assays) or <1TCID(50)ml(-1) (quantitative real-time assays). Mie scattering simulations were able to predict the shape of the PRRSV standard curve, indicating that any non-linearity of the standard curve can be interpreted purely as an optical phenomenon. Each assay took less than 5min. A strong correlation could be found between RT-PCR and this method for the lung tissue samples, even though their respective detection mechanisms are different fundamentally (nucleic acids for RT-PCR and virus antigens for light scattering immunoagglutination assay). Several different dilution factors were also tested for tissue samples, and 1/10 and 1/100 were found to be usable. If the microfluidic chips are used only once (i.e. without re-using them), both superior sensitivity and satisfactory specificity can be demonstrated. Specificity studies revealed the presence of Type II PRRSV and non-presence of Type I PRRSV and that the microfluidic chip assay could detect Type II North American strain of PRRSV for the animals tested. This work demonstrates the potential of the Mie

  14. On-chip mitochondrial assay microfluidic devices and protein nanopore/nanotube hybrid transistor

    NASA Astrophysics Data System (ADS)

    Lim, Taesun

    Tremendous efforts to understand the cause, mechanism of development and the way to treat various diseases as well as an early diagnosis have been made so far and people are still working hardly on these researches. Even now, countless people are suffering from diseases such as Alzhemer's disease, Parkinson's disease, diabetes and cancer without knowing clues to cure their diseases completely. Generally speaking, we still have a long way to go through to comprehensively figure out these our long-lasting homeworks. One of possible solutions is to merge current advanced technology and science together to find a powerful synergetic effect for a specific purpose that can be tailored depending on user's need. Here this research tried to put nanotechnology and biological science together to find a way to resolve current challenges by developing a new generation of the analytical sensing device. Mitochondrial functions and biological roles in regulating life and death control will be discussed indicating mitochondrion is a crucial organism to monitor to obtain important information regarding degenerative diseases and aging process. On-chip mitochondrial functional assay microsensor that could facilitate the mitochondrial evaluation will be extensively demonstrated and discussed in both technical and biological perspectives. The novel fusion technological approach will be demonstrated by combining artificial cell membrane with carbon nanotube electronics to interrogate interactions between biomolecules and electronic circuitries. In addition, molecular dynamics at the cell membrane could be investigated closely which can help understand the cell-cell communication and the regulation of ion transport.

  15. Targeted Interleukin-10 Nanotherapeutics Developed with a Microfluidic Chip Enhance Resolution of Inflammation in Advanced Atherosclerosis.

    PubMed

    Kamaly, Nazila; Fredman, Gabrielle; Fojas, Jhalique Jane R; Subramanian, Manikandan; Choi, Won Ii; Zepeda, Katherine; Vilos, Cristian; Yu, Mikyung; Gadde, Suresh; Wu, Jun; Milton, Jaclyn; Carvalho Leitao, Renata; Rosa Fernandes, Livia; Hasan, Moaraj; Gao, Huayi; Nguyen, Vance; Harris, Jordan; Tabas, Ira; Farokhzad, Omid C

    2016-05-24

    Inflammation is an essential protective biological response involving a coordinated cascade of signals between cytokines and immune signaling molecules that facilitate return to tissue homeostasis after acute injury or infection. However, inflammation is not effectively resolved in chronic inflammatory diseases such as atherosclerosis and can lead to tissue damage and exacerbation of the underlying condition. Therapeutics that dampen inflammation and enhance resolution are currently of considerable interest, in particular those that temper inflammation with minimal host collateral damage. Here we present the development and efficacy investigations of controlled-release polymeric nanoparticles incorporating the anti-inflammatory cytokine interleukin 10 (IL-10) for targeted delivery to atherosclerotic plaques. Nanoparticles were nanoengineered via self-assembly of biodegradable polyester polymers by nanoprecipitation using a rapid micromixer chip capable of producing nanoparticles with retained IL-10 bioactivity post-exposure to organic solvent. A systematic combinatorial approach was taken to screen nanoparticles, resulting in an optimal bioactive formulation from in vitro and ex vivo studies. The most potent nanoparticle termed Col-IV IL-10 NP22 significantly tempered acute inflammation in a self-limited peritonitis model and was shown to be more potent than native IL-10. Furthermore, the Col-IV IL-10 nanoparticles prevented vulnerable plaque formation by increasing fibrous cap thickness and decreasing necrotic cores in advanced lesions of high fat-fed LDLr(-/-) mice. These results demonstrate the efficacy and pro-resolving potential of this engineered nanoparticle for controlled delivery of the potent IL-10 cytokine for the treatment of atherosclerosis.

  16. Comparison of roll-to-roll replication approaches for microfluidic and optical functions in lab-on-a-chip diagnostic devices

    NASA Astrophysics Data System (ADS)

    Brecher, Christian; Baum, Christoph; Bastuck, Thomas

    2015-03-01

    Economically advantageous microfabrication technologies for lab-on-a-chip diagnostic devices substituting commonly used glass etching or injection molding processes are one of the key enablers for the emerging market of microfluidic devices. On-site detection in fields of life sciences, point of care diagnostics and environmental analysis requires compact, disposable and highly functionalized systems. Roll-to-roll production as a high volume process has become the emerging fabrication technology for integrated, complex high technology products within recent years (e.g. fuel cells). Differently functionalized polymer films enable researchers to create a new generation of lab-on-a-chip devices by combining electronic, microfluidic and optical functions in multilayer architecture. For replication of microfluidic and optical functions via roll-to-roll production process competitive approaches are available. One of them is to imprint fluidic channels and optical structures of micro- or nanometer scale from embossing rollers into ultraviolet (UV) curable lacquers on polymer substrates. Depending on dimension, shape and quantity of those structures there are alternative manufacturing technologies for the embossing roller. Ultra-precise diamond turning, electroforming or casting polymer materials are used either for direct structuring or manufacturing of roller sleeves. Mastering methods are selected for application considering replication quality required and structure complexity. Criteria for the replication quality are surface roughness and contour accuracy. Structure complexity is evaluated by shapes producible (e.g. linear, circular) and aspect ratio. Costs for the mastering process and structure lifetime are major cost factors. The alternative replication approaches are introduced and analyzed corresponding to the criteria presented. Advantages and drawbacks of each technology are discussed and exemplary applications are presented.

  17. Microfluidic cell culture chip with multiplexed medium delivery and efficient cell/scaffold loading mechanisms for high-throughput perfusion 3-dimensional cell culture-based assays.

    PubMed

    Huang, Song-Bin; Wu, Min-Hsien; Wang, Shih-Siou; Lee, Gwo-Bin

    2011-06-01

    This study reports a microfluidic cell culture chip consisting of 48 microbioreactors for high-throughput perfusion 3-dimensional (3-D) cell culture-based assays. Its advantages include the capability for multiplexed and backflow-free medium delivery, and both efficient and high-throughput micro-scale, 3-D cell culture construct loading. In this work, the microfluidic cell culture chip is fabricated using two major processes, specifically, a computer-numerical-controlled (CNC) mold machining process and a polydimethylsiloxane (PDMS) replication process. The chip is composed of micropumps, microbioreactors, connecting microchannels and a cell/agarose scaffold loading mechanism. The performance of the new pneumatic micropumps and the cell/agarose scaffold loading mechanism has been experimentally evaluated. The experimental results show that this proposed multiplexed medium-pumping design is able to provide a uniform pumping rate ranging from 1.5 to 298.3 μl hr(-1) without any fluid backflow and the resultant medium contamination. In addition, the simple cell/agarose loading method has been proven to be able to load the 3-D cell culture construct uniformly and efficiently in all 48 microbioreactors investigated. Furthermore, a micro-scale, perfusion, 3-D cell culture-based assay has been successfully demonstrated using this proposed cell culture chip. The experimental results are also compared to a similar evaluation using a conventional static 3-D cell culture with a larger scale culture. It is concluded that the choice of a cell culture format can influence assay results. As a whole, because of the inherent advantages of a miniaturized perfusion 3-D cell culture assay, the cell culture chip not only can provide a stable, well-defined and more biologically-meaningful culture environment, but it also features a low consumption of research resources. Moreover, due to the integrated medium pumping mechanism and the simple cell/agarose loading method, this chip is

  18. Control of initiation, rate, and routing of spontaneous capillary-driven flow of liquid droplets through microfluidic channels on SlipChip.

    PubMed

    Pompano, Rebecca R; Platt, Carol E; Karymov, Mikhail A; Ismagilov, Rustem F

    2012-01-24

    This Article describes the use of capillary pressure to initiate and control the rate of spontaneous liquid-liquid flow through microfluidic channels. In contrast to flow driven by external pressure, flow driven by capillary pressure is dominated by interfacial phenomena and is exquisitely sensitive to the chemical composition and geometry of the fluids and channels. A stepwise change in capillary force was initiated on a hydrophobic SlipChip by slipping a shallow channel containing an aqueous droplet into contact with a slightly deeper channel filled with immiscible oil. This action induced spontaneous flow of the droplet into the deeper channel. A model predicting the rate of spontaneous flow was developed on the basis of the balance of net capillary force with viscous flow resistance, using as inputs the liquid-liquid surface tension, the advancing and receding contact angles at the three-phase aqueous-oil-surface contact line, and the geometry of the devices. The impact of contact angle hysteresis, the presence or absence of a lubricating oil layer, and adsorption of surface-active compounds at liquid-liquid or liquid-solid interfaces were quantified. Two regimes of flow spanning a 10(4)-fold range of flow rates were obtained and modeled quantitatively, with faster (mm/s) flow obtained when oil could escape through connected channels as it was displaced by flowing aqueous solution, and slower (micrometer/s) flow obtained when oil escape was mostly restricted to a micrometer-scale gap between the plates of the SlipChip ("dead-end flow"). Rupture of the lubricating oil layer (reminiscent of a Cassie-Wenzel transition) was proposed as a cause of discrepancy between the model and the experiment. Both dilute salt solutions and complex biological solutions such as human blood plasma could be flowed using this approach. We anticipate that flow driven by capillary pressure will be useful for the design and operation of flow in microfluidic applications that do not

  19. Control of initiation, rate, and routing of spontaneous capillary-driven flow of liquid droplets through microfluidic channels on SlipChip

    PubMed Central

    Pompano, Rebecca R.; Platt, Carol E.; Karymov, Mikhail A.

    2012-01-01

    This paper describes the use of capillary pressure to initiate and control the rate of spontaneous liquid-liquid flow through microfluidic channels. In contrast to flow driven by external pressure, flow driven by capillary pressure is dominated by interfacial phenomena and is exquisitely sensitive to the chemical composition and geometry of the fluids and channels. A step-wise change in capillary force was initiated on a hydrophobic SlipChip by slipping a shallow channel containing an aqueous droplet into contact with a slightly deeper channel filled with immiscible oil. This action induced spontaneous flow of the droplet into the deeper channel. A model predicting the rate of spontaneous flow was developed based on the balance of net capillary force with viscous flow resistance, using as inputs the liquid-liquid surface tension, the advancing and receding contact angles at the three-phase aqueous-oil-surface contact line, and the geometry of the devices. The impact of contact angle hysteresis, the presence or absence of a lubricating oil layer, and adsorption of surface-active compounds at liquid-liquid or liquid-solid interfaces were quantified. Two regimes of flow spanning a 104-fold range of flow rates were obtained and modeled quantitatively, with faster (mm/s) flow obtained when oil could escape through connected channels as it was displaced by flowing aqueous solution, and slower (micrometer/s) flow obtained when oil escape was mostly restricted to a μm-scale gap between the plates of the SlipChip (“dead-end flow”). Rupture of the lubricating oil layer (reminiscent of a Cassie-Wenzel transition) was proposed as a cause of discrepancy between the model and the experiment. Both dilute salt solutions and complex biological solutions such as human blood plasma could be flowed using this approach. We anticipate that flow driven by capillary pressure will be useful for design and operation of flow in microfluidic applications that do not require external

  20. Surface plasmon resonance imaging for affinity analysis of aptamer-protein interactions with PDMS microfluidic chips.

    PubMed

    Wang, Zhuangzhi; Wilkop, Thomas; Xu, Danke; Dong, Yi; Ma, Guangyu; Cheng, Quan

    2007-10-01

    We report on the use of PDMS multichannels for affinity studies of DNA aptamer-human Immunoglobulin E (IgE) interactions by surface plasmon resonance imaging (SPRi). The sensing surface was prepared with thiol-terminated aptamers through a self-assembling process in the PDMS channels defined on a gold substrate. Cysteamine was codeposited with the thiol aptamers to promote proper spatial arrangement of the aptamers and thus maintain their optimal binding efficiencies. Four aptamers with different nucleic acid sequences were studied to test their interaction affinity toward IgE, and the results confirmed that aptamer I (5'-SH-GGG GCA CGT TTA TCC GTC CCT CCT AGT GGC GTG CCC C-3') has the strongest binding affinity. Control experiments were conducted with a PEG-functionalized surface and IgG was used to replace IgE in order to verify the selective binding of aptamer I to the IgE molecules. A linear concentration-dependent relationship between IgE and aptamer I was obtained, and a 2-nM detection limit was achieved. SPRi data were further analyzed by global fitting, and the dissociation constant of aptamer I-IgE complex was found to be 2.7 x 10(-7) M, which agrees relatively well with the values reported in the literature. Aptamer affinity screening by SPR imaging demonstrates marked advantages over competing methods because it does not require labeling, can be used in real-time, and is potentially high-throughput. The ability to provide both qualitative and quantitative results on a multichannel chip further establishes SPRi as a powerful tool for the study of biological interactions in a multiplexed format.

  1. The study of energy metabolism in bladder cancer cells in co-culture conditions using a microfluidic chip

    PubMed Central

    Xu, Xiao-Dong; Shao, Shi-Xiu; Cao, Yan-Wei; Yang, Xue-Cheng; Shi, Hao-Qing; Wang, You-Lin; Xue, Sen-Yao; Wang, Xin-Sheng; Niu, Hai-Tao

    2015-01-01

    Objectives: This study aimed to systematically analyze changes in mitochondrial-related protein expression in bladder cancer cells and tumor-associated fibroblasts and to investigate the characteristics of bladder cancer cell energy metabolism. Methods: In this study, we utilized the following techniques to achieve the objectives: (1) a co-culture system of bladder tumor cells and fibroblasts was built using a microfluidic chip as a three-dimensional culture system; (2) the concentration of lactic acid in the medium from the different groups was determined using an automatic micro-plate reader; (3) a qualitative analysis of mitochondria-related protein expression was performed by immunofluorescent staining; and (4) a quantitative analysis of mitochondrial-associated protein expression was conducted via Western blot. SPSS software was utilized to analyze the data. Results: (1) Determination of lactic acid concentration: The lactic acid concentration was determined to be highest in the experimental group, followed by the T24 cell control group and then the fibroblast control group. (2) Qualitative results: In the control group, the mitochondrial-related protein fluorescence intensity was higher in the fibroblasts compared with the cancer cells, and the fluorescence intensity of the fibroblasts was reduced compared with the experimental group. The mitochondrial-related protein fluorescence intensity of the cancer cells was higher in the experimental group compared with the control group, and the opposite results were obtained with the fibroblasts. (3) Quantitative results: The expression of mitochondria-related proteins was higher in fibroblasts compared with cancer cells in the control group, and the opposite results were obtained in the experimental group (P<0.05). The expression of mitochondria-related proteins was increased in cancer cells in the experimental group compared with the control group; the opposite results were observed for the fibroblasts (P<0

  2. A Microfluidics-HPLC/Differential Mobility Spectrometer Macromolecular Detection System for Human and Robotic Missions

    NASA Astrophysics Data System (ADS)

    Coy, S. L.; Killeen, K.; Han, J.; Eiceman, G. A.; Kanik, I.; Kidd, R. D.

    2011-03-01

    Our goal is to develop a unique, miniaturized, solute analyzer based on microfluidics technology. The analyzer consists of an integrated microfluidics High Performance Liquid Chromatographic chip/Differential Mobility Spectrometer (HPLC-chip/DMS) detection system.

  3. Microfluidic Platforms for on-chip Formulation and Small-Angle x-ray Analysis of the Phase Behavior of Lipid/Water Mixtures

    SciTech Connect

    Khvostichenko, Daria S.; Perry, Sarah L.; Kondrashkina, Elena; Guha, Sudipto; Brister, Keith; Kenis, Paul J.A.

    2012-03-27

    We present a microfluidic platform for on-chip formulation and X-ray analysis of lipidic mesophases formed upon mixing lipids and water. The platform is designed to study the effect of detergents on the phase behavior of lipid/water mixtures. The platform allows automated preparation of multiple samples of different composition from stock solutions and subsequent on-chip small-angle X-ray diffraction (SAXS) data collection. To ensure X-ray transparency of the platform we used thin layers of cyclic olefin copolymer (COC) and PDMS. The viability of the platform is demonstrated by mapping out a section of the phase diagram for lipid monoolein mixed with solutions of detergent {beta}-octylglucoside. The platform reported here is a viable alternative to the traditional method of establishing phase diagrams for lipid/solution mixtures. Compared to the conventional approach, a significantly smaller amount of sample is required for mapping phase diagrams of lipidic mesophases and samples of various compositions are prepared automatically. In ongoing work we are using these chips to rapidly determine the phase behavior of a range of lipids to establish their suitability for membrane protein crystallization, especially with respect to their sensitivity to detergent concentration.

  4. 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.

  5. Spectroscopic imaging system for high-throughput viability assessment of ovarian spheroids or microdissected tumor tissues (MDTs) in a microfluidic chip

    NASA Astrophysics Data System (ADS)

    St-Georges-Robillard, A.; Masse, M.; Kendall-Dupont, J.; Strupler, M.; Patra, B.; Jermyn, M.; Mes-Masson, A.-M.; Leblond, F.; Gervais, T.

    2016-02-01

    There is a growing effort in the biomicrosystems community to develop a personalized treatment response assay for cancer patients using primary cells, patient-derived spheroids, or live tissues on-chip. Recently, our group has developed a technique to cut tumors in 350 μm diameter microtissues and keep them alive on-chip, enabling multiplexed in vitro drug assays on primary tumor tissue. Two-photon microscopy, confocal microscopy and flow cytometry are the current standard to assay tissue chemosensitivity on-chip. While these techniques provide microscopic and molecular information, they are not adapted for high-throughput analysis of microtissues. We present a spectroscopic imaging system that allows rapid quantitative measurements of multiple fluorescent viability markers simultaneously by using a liquid crystal tunable filter to record fluorescence and transmittance spectra. As a proof of concept, 24 spheroids composed of ovarian cancer cell line OV90 were formed in a microfluidic chip, stained with two live cell markers (CellTrackerTM Green and Orange), and imaged. Fluorescence images acquired were normalized to the acquisition time and gain of the camera, dark noise was removed, spectral calibration was applied, and spatial uniformity was corrected. Spectral un-mixing was applied to separate each fluorophore's contribution. We have demonstrated that rapid and simultaneous viability measurements on multiple spheroids can be achieved, which will have a significant impact on the prediction of a tumor's response to multiple treatment options. This technique may be applied as well in drug discovery to assess the potential of a drug candidate directly on human primary tissue.

  6. A fully battery-powered inexpensive spectrophotometric system for high-sensitivity point-of-care analysis on a microfluidic chip.

    PubMed

    Dou, Maowei; Lopez, Juan; Rios, Misael; Garcia, Oscar; Xiao, Chuan; Eastman, Michael; Li, XiuJun

    2016-06-21

    A cost-effective b[combining low line]a[combining low line]ttery-powered s[combining low line]pectrophotometric s[combining low line]ystem (BASS) was developed for quantitative point-of-care (POC) analysis on a microfluidic chip. By using methylene blue as a model analyte, we first compared the performance of the BASS with a commercial spectrophotometric system, and further applied the BASS for loop-mediated isothermal amplification (LAMP) detection and subsequent quantitative nucleic acid analysis which exhibited a comparable limit of detection to that of Nanodrop. Compared to the commercial spectrophotometric system, our spectrophotometric system is lower-cost, consumes less reagents, and has higher detection sensitivity. Most importantly, it does not rely on external power supplies. All these features make our spectrophotometric system highly suitable for a variety of POC analyses, such as field detection.

  7. Room-temperature serial crystallography using a kinetically optimized microfluidic device for protein crystallization and on-chip X-ray diffraction

    PubMed Central

    Heymann, Michael; Opthalage, Achini; Wierman, Jennifer L.; Akella, Sathish; Szebenyi, Doletha M. E.; Gruner, Sol M.; Fraden, Seth

    2014-01-01

    An emulsion-based serial crystallographic technology has been developed, in which nanolitre-sized droplets of protein solution are encapsulated in oil and stabilized by surfactant. Once the first crystal in a drop is nucleated, the small volume generates a negative feedback mechanism that lowers the supersaturation. This mechanism is exploited to produce one crystal per drop. Diffraction data are measured, one crystal at a time, from a series of room-temperature crystals stored on an X-ray semi-transparent microfluidic chip, and a 93% complete data set is obtained by merging single diffraction frames taken from different unoriented crystals. As proof of concept, the structure of glucose isomerase was solved to 2.1 Å, demonstrating the feasibility of high-throughput serial X-ray crystallography using synchrotron radiation. PMID:25295176

  8. Efficient radiosynthesis of 3′-deoxy-3′-[18F]fluorothymidine using electrowetting-on-dielectric digital microfluidic chip

    PubMed Central

    Javed, Muhammad Rashed; Chen, Supin; Kim, Hee-Kwon; Wei, Liu; Czernin, Johannes; Kim, Chang-Jin “CJ”; van Dam, R. Michael; Keng, Pei Yuin

    2015-01-01

    Access to diverse PET tracers for preclinical and clinical research remains a major obstacle to research in cancer and other diseases research. The prohibitive cost and limited availability of tracers could be alleviated by microfluidic radiosynthesis technologies combined with high-yield microscale radiosynthetic method. In this report, we demonstrate the multistep synthesis of 3′-deoxy-3′-[18F]fluorothymidine ([18F]FLT) with high yield on an electrowetting on dielectric (EWOD) microfluidic radiosynthesizer, previously developed in our group. We have identified and established several parameters that are most critical in the microscale radiosynthesis such as the reaction time, reagent concentration, and molar ratios, to successfully synthesize [18F]FLT in this compact platform. Methods [18F]FLT was synthesized from the 3-N-Boc-1-[5-O-(4,4′-dimethoxytrityl)-3-O-nosyl-2-deoxy-β-d-lyxofuranosyl] thymine precursor on the EWOD chip starting from the first solvent exchange and [18F]fluoride ion activation step to the final deprotection step. The fluorination reaction was performed in a mixture of thexyl alcohol and DMSO. The crude product after deprotection was collected from the chip and purified on a custom-made solid phase extraction (SPE) cartridge and subjected to quality control testing. The purified [18F]FLT was suitable for microPET studies in multiple nude mice xenografted with the A431 carcinoma cell line. Results [18F]FLT was successfully synthesized on the EWOD microdevice coupled with an off-chip SPE purification with a decayed-corrected radiochemical yield of 63±5% (n=5) and passed all of the quality control test required by the United States Pharmacopeia for radiotracers to be injected into humans. We have successfully demonstrated the synthesis of several batches of [18F]FLT on EWOD starting with ∼ 333 MBq of radioactivity and obtained up to 52 MBq (non-decay corrected) of [18F]FLT upon cartridge purification. The specific activity of two

  9. On-Chip Titration of an Anticoagulant Argatroban and Determination of the Clotting Time within Whole Blood or Plasma Using a Plug-Based Microfluidic System

    PubMed Central

    Song, Helen; Li, Hung-Wing; Munson, Matthew S.; Van Ha, Thuong G.; Ismagilov, Rustem F.

    2006-01-01

    This paper describes extending plug-based microfluidics to handling complex biological fluids such as blood, solving the problem of injecting additional reagents into plugs, and applying this system to measuring of clotting time in small volumes of whole blood and plasma. Plugs are droplets transported through microchannels by fluorocarbon fluids. A plug-based microfluidic system was developed to titrate an anticoagulant (argatroban) into blood samples and to measure the clotting time using the activated partial thromboplastin time (APTT) test. To carry out these experiments, the following techniques were developed for a plug-based system: (i) using Teflon AF coating on the microchannel wall to enable formation of plugs containing blood and transport of the solid fibrin clots within plugs, (ii) using a hydrophilic glass capillary to enable reliable merging of a reagent from an aqueous stream into plugs, (iii) using bright-field microscopy to detect the formation of a fibrin clot within plugs and using fluorescent microscopy to detect the production of thrombin using a fluorogenic substrate, and (iv) titration of argatroban (0–1.5 μg/mL) into plugs and measurement of the resulting APTTs at room temperature (23 °C) and physiological temperature (37 °C). APTT measurements were conducted with normal pooled plasma (platelet-poor plasma) and with donor’s blood samples (both whole blood and platelet-rich plasma). APTT values and APTT ratios measured by the plug-based microfluidic device were compared to the results from a clinical laboratory at 37 °C. APTT obtained from the on-chip assay were about double those from the clinical laboratory but the APTT ratios from these two methods agreed well with each other. PMID:16841902

  10. On-chip titration of an anticoagulant argatroban and determination of the clotting time within whole blood or plasma using a plug-based microfluidic system.

    PubMed

    Song, Helen; Li, Hung-Wing; Munson, Matthew S; Van Ha, Thuong G; Ismagilov, Rustem F

    2006-07-15

    This paper describes extending plug-based microfluidics to handling complex biological fluids such as blood, solving the problem of injecting additional reagents into plugs, and applying this system to measuring of clotting time in small volumes of whole blood and plasma. Plugs are droplets transported through microchannels by fluorocarbon fluids. A plug-based microfluidic system was developed to titrate an anticoagulant (argatroban) into blood samples and to measure the clotting time using the activated partial thromboplastin time (APTT) test. To carry out these experiments, the following techniques were developed for a plug-based system: (i) using Teflon AF coating on the microchannel wall to enable formation of plugs containing blood and transport of the solid fibrin clots within plugs, (ii) using a hydrophilic glass capillary to enable reliable merging of a reagent from an aqueous stream into plugs, (iii) using bright-field microscopy to detect the formation of a fibrin clot within plugs and using fluorescent microscopy to detect the production of thrombin using a fluorogenic substrate, and (iv) titration of argatroban (0-1.5 microg/mL) into plugs and measurement of the resulting APTTs at room temperature (23 degrees C) and physiological temperature (37 degrees C). APTT measurements were conducted with normal pooled plasma (platelet-poor plasma) and with donor's blood samples (both whole blood and platelet-rich plasma). APTT values and APTT ratios measured by the plug-based microfluidic device were compared to the results from a clinical laboratory at 37 degrees C. APTT obtained from the on-chip assay were about double those from the clinical laboratory but the APTT ratios from these two methods agreed well with each other.

  11. Combination of capillary micellar liquid chromatography with on-chip microfluidic chemiluminescence detection for direct analysis of buspirone in human plasma.

    PubMed

    Al Lawati, Haider A J; Kadavilpparampu, Afsal Mohammed; Suliman, FakhrEldin O

    2014-09-01

    Microfluidic based chemiluminescence (CL) detector having novel channel design for enhanced mixing has been developed and investigated in terms of its applicability with micellar mode of liquid chromatography (MLC). The newly developed detector was found to be highly sensitive and an alternative detection technique to combine with capillary MLC. This combination was successfully employed for direct detection of a model analyte using Ru(III)-peroxydisulphate CL system. The selected analyte, buspirone hydrochloride (BUS), was detected selectively at therapeutic concentration levels in human plasma without any sample pretreatment. By incorporating eight flow split units within the spiral channel of microfluidic chip, an enhancement of 140% in CL emission was observed. We also evaluated the effect of non- ionic surfactant, Brij-35, which used as mobile phase modifier in MLC, on CL emission. The CL signal was improved by 52% compared to aqueous-organic mobile phase combinations. Various parameters influencing the micellar chromatographic performance and the CL emission were optimized. This allowed highly sensitive analysis of BUS with limit of detection (LOD) of 0.27 ng mL(-1) (3σ/s) and limit of quantification (LOQ) of 0.89 ng mL(-1) (10σ/s). The analyte recovery from human plasma at three different concentration level ranges from 88% to 96% (RSD 1.9-5.3%). The direct analysis of BUS in human plasma was achieved within 6 min. Therefore, combining microfluidic CL detection with micellar mode of separation is an efficient, cost-effective and highly sensitive technique that can utilize MLC in its full capacity for various bioanalytical procedures.

  12. Multi-layered, membrane-integrated microfluidics based on replica molding of a thiol-ene epoxy thermoset for organ-on-a-chip applications.

    PubMed

    Sticker, Drago; Rothbauer, Mario; Lechner, Sarah; Hehenberger, Marie-Therese; Ertl, Peter

    2015-12-21

    In this study we have investigated a photosensitive thermoset (OSTEMER 322-40) as a complementary material to readily fabricate complex multi-layered microdevices for applications in life science. Simple, versatile and robust fabrication of multifunctional microfluidics is becoming increasingly important for the development of customized tissue-, organ- and body-on-a-chip systems capable of mimicking tissue interfaces and biological barriers. In the present work key material properties including optical properties, vapor permeability, hydrophilicity and biocompatibility are evaluated for cell-based assays using fibroblasts, endothelial cells and mesenchymal stem cells. The excellent bonding strength of the OSTEMER thermoset to flexible fluoropolymer (FEP) sheets and poly(dimethylsiloxane) (PDMS) membranes further allows for the fabrication of integrated microfluidic components such as membrane-based microdegassers, microvalves and micropumps. We demonstrate the application of multi-layered, membrane-integrated microdevices that consist of up to seven layers and three membranes that specially confine and separate vascular cells from the epithelial barrier and 3D tissue structures.

  13. Recent Progress of Microfluidics in Translational Applications

    PubMed Central

    Liu, Zongbin; Han, Xin

    2016-01-01

    Microfluidics, featuring microfabricated structures, is a technology for manipulating fluids at the micrometer scale. The small dimension and flexibility of microfluidic systems are ideal for mimicking molecular and cellular microenvironment, and show great potential in translational research and development. Here, the recent progress of microfluidics in biological and biomedical applications, including molecular analysis, cellular analysis, and chip-based material delivery and biomimetic design is presented. The potential future developments in the translational microfluidics field are also discussed. PMID:27091777

  14. Recent Progress of Microfluidics in Translational Applications.

    PubMed

    Liu, Zongbin; Han, Xin; Qin, Lidong

    2016-04-20

    Microfluidics, featuring microfabricated structures, is a technology for manipulating fluids at the micrometer scale. The small dimension and flexibility of microfluidic systems are ideal for mimicking molecular and cellular microenvironment, and show great potential in translational research and development. Here, the recent progress of microfluidics in biological and biomedical applications, including molecular analysis, cellular analysis, and chip-based material delivery and biomimetic design is presented. The potential future developments in the translational microfluidics field are also discussed.

  15. Heat Transfer and Friction Characteristics of the Microfluidic Heat Sink with Variously-Shaped Ribs for Chip Cooling

    PubMed Central

    Wang, Gui-Lian; Yang, Da-Wei; Wang, Yan; Niu, Di; Zhao, Xiao-Lin; Ding, Gui-Fu

    2015-01-01

    This paper experimentally and numerically investigated the heat transfer and friction characteristics of microfluidic heat sinks with variously-shaped micro-ribs, i.e., rectangular, triangular and semicircular ribs. The micro-ribs were fabricated on the sidewalls of microfluidic channels by a surface-micromachining micro-electro-mechanical system (MEMS) process and used as turbulators to improve the heat transfer rate of the microfluidic heat sink. The results indicate that the utilizing of micro-ribs provides a better heat transfer rate, but also increases the pressure drop penalty for microchannels. Furthermore, the heat transfer and friction characteristics of the microchannels are strongly affected by the rib shape. In comparison, the triangular ribbed microchannel possesses the highest Nusselt number and friction factor among the three rib types. PMID:25912351

  16. Heat transfer and friction characteristics of the microfluidic heat sink with variously-shaped ribs for chip cooling.

    PubMed

    Wang, Gui-Lian; Yang, Da-Wei; Wang, Yan; Niu, Di; Zhao, Xiao-Lin; Ding, Gui-Fu

    2015-04-22

    This paper experimentally and numerically investigated the heat transfer and friction characteristics of microfluidic heat sinks with variously-shaped micro-ribs, i.e., rectangular, triangular and semicircular ribs. The micro-ribs were fabricated on the sidewalls of microfluidic channels by a surface-micromachining micro-electro-mechanical system (MEMS) process and used as turbulators to improve the heat transfer rate of the microfluidic heat sink. The results indicate that the utilizing of micro-ribs provides a better heat transfer rate, but also increases the pressure drop penalty for microchannels. Furthermore, the heat transfer and friction characteristics of the microchannels are strongly affected by the rib shape. In comparison, the triangular ribbed microchannel possesses the highest Nusselt number and friction factor among the three rib types.

  17. Microfluidic on-chip capture-cycloaddition reaction to reversibly immobilize small molecules or multi-component structures for biosensor applications.

    PubMed

    Tassa, Carlos; Liong, Monty; Hilderbrand, Scott; Sandler, Jason E; Reiner, Thomas; Keliher, Edmund J; Weissleder, Ralph; Shaw, Stanley Y

    2013-09-23

    Methods for rapid surface immobilization of bioactive small molecules with control over orientation and immobilization density are highly desirable for biosensor and microarray applications. In this Study, we use a highly efficient covalent bioorthogonal [4+2] cycloaddition reaction between trans-cyclooctene (TCO) and 1,2,4,5-tetrazine (Tz) to enable the microfluidic immobilization of TCO/Tz-derivatized molecules. We monitor the process in real-time under continuous flow conditions using surface plasmon resonance (SPR). To enable reversible immobilization and extend the experimental range of the sensor surface, we combine a non-covalent antigen-antibody capture component with the cycloaddition reaction. By alternately presenting TCO or Tz moieties to the sensor surface, multiple capture-cycloaddition processes are now possible on one sensor surface for on-chip assembly and interaction studies of a variety of multi-component structures. We illustrate this method with two different immobilization experiments on a biosensor chip; a small molecule, AP1497 that binds FK506-binding protein 12 (FKBP12); and the same small molecule as part of an immobilized and in situ-functionalized nanoparticle.

  18. 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.

  19. Real-time microfluidic recombinase polymerase amplification for the toxin B gene of Clostridium difficile on a SlipChip platform.

    PubMed

    Tsaloglou, M-N; Watson, R J; Rushworth, C M; Zhao, Y; Niu, X; Sutton, J M; Morgan, H

    2015-01-07

    Clostridium difficile is one of the key bacterial pathogens that cause infectious diarrhoea both in the developed and developing world. Isothermal nucleic acid amplification methods are increasingly used for identification of toxinogenic infection by clinical labs. For this purpose, we developed a low-cost microfluidic platform based on the SlipChip concept and implemented real-time isothermal recombinase polymerase amplification (RPA). The on-chip RPA assay targets the Clostridium difficile toxin B gene (tcdB) coding for toxin B, one of the proteins responsible for bacterial toxicity. The device was fabricated in clear acrylic using rapid prototyping methods. It has six replicate 500 nL reaction wells as well as two sets of 500 nL control wells. The reaction can be monitored in real-time using exonuclease fluorescent probes with an initial sample volume of as little as 6.4 μL. We demonstrated a limit of detection of 1000 DNA copies, corresponding to 1 fg, at a time-to-result of <20 minutes. This miniaturised platform for pathogen detection has potential for use in resource-limited environments or at the point-of-care because of its ease of use and low cost, particularly if combined with preserved reagents.

  20. MEMS in microfluidic channels.

    SciTech Connect

    Ashby, Carol Iris Hill; Okandan, Murat; Michalske, Terry A.; Sounart, Thomas L.; Matzke, Carolyn M.

    2004-03-01

    Microelectromechanical systems (MEMS) comprise a new class of devices that include various forms of sensors and actuators. Recent studies have shown that microscale cantilever structures are able to detect a wide range of chemicals, biomolecules or even single bacterial cells. In this approach, cantilever deflection replaces optical fluorescence detection thereby eliminating complex chemical tagging steps that are difficult to achieve with chip-based architectures. A key challenge to utilizing this new detection scheme is the incorporation of functionalized MEMS structures within complex microfluidic channel architectures. The ability to accomplish this integration is currently limited by the processing approaches used to seal lids on pre-etched microfluidic channels. This report describes Sandia's first construction of MEMS instrumented microfluidic chips, which were fabricated by combining our leading capabilities in MEMS processing with our low-temperature photolithographic method for fabricating microfluidic channels. We have explored in-situ cantilevers and other similar passive MEMS devices as a new approach to directly sense fluid transport, and have successfully monitored local flow rates and viscosities within microfluidic channels. Actuated MEMS structures have also been incorporated into microfluidic channels, and the electrical requirements for actuation in liquids have been quantified with an elegant theory. Electrostatic actuation in water has been accomplished, and a novel technique for monitoring local electrical conductivities has been invented.

  1. Electrorheological fluid and its applications in microfluidics.

    PubMed

    Wang, Limu; Gong, Xiuqing; Wen, Weijia

    2011-01-01

    Microfluidics is a low-cost technique for fast-diagnosis and microsynthesis. Within a decade it might become the foundation of point-of-care and lab-on-a-chip applications. With microfluidic chips, high-throughput sample screening and information processing are made possible. The picoliter droplet runs in microfluidic chips are ideal miniaturized vessels for microdetection and microsynthesis. Meanwhile, individual manipulation of microdroplets remains a challenge: the shortcomings in automatic, reliable, and scalable methods for logic control prevent further integration of microfluidic applications. The giant electrorheological fluid (GERF), which is a kind of "smart" colloid, has tunable viscosity under the influence of external electric field. Therefore, GERF is introduced as the active controlling medium, with real-time response in on-chip fluid control. This review article introduces the working principles and fabrication methods of different types of electrorheological fluid, and extensively describes the strategies of GERF-assisted microfluidic controlling schemes.

  2. Punch card programmable microfluidics.

    PubMed

    Korir, George; Prakash, Manu

    2015-01-01

    Small volume fluid handling in single and multiphase microfluidics provides a promising strategy for efficient bio-chemical assays, low-cost point-of-care diagnostics and new approaches to scientific discoveries. However multiple barriers exist towards low-cost field deployment of programmable microfluidics. Incorporating multiple pumps, mixers and discrete valve based control of nanoliter fluids and droplets in an integrated, programmable manner without additional required external components has remained elusive. Combining the idea of punch card programming with arbitrary fluid control, here we describe a self-contained, hand-crank powered, multiplex and robust programmable microfluidic platform. A paper tape encodes information as a series of punched holes. A mechanical reader/actuator reads these paper tapes and correspondingly executes operations onto a microfluidic chip coupled to the platform in a plug-and-play fashion. Enabled by the complexity of codes that can be represented by a series of holes in punched paper tapes, we demonstrate independent control of 15 on-chip pumps with enhanced mixing, normally-closed valves and a novel on-demand impact-based droplet generator. We demonstrate robustness of operation by encoding a string of characters representing the word "PUNCHCARD MICROFLUIDICS" using the droplet generator. Multiplexing is demonstrated by implementing an example colorimetric water quality assays for pH, ammonia, nitrite and nitrate content in different water samples. With its portable and robust design, low cost and ease-of-use, we envision punch card programmable microfluidics will bring complex control of microfluidic chips into field-based applications in low-resource settings and in the hands of children around the world.

  3. Cell manipulation in microfluidics.

    PubMed

    Yun, Hoyoung; Kim, Kisoo; Lee, Won Gu

    2013-06-01

    Recent advances in the lab-on-a-chip field in association with nano/microfluidics have been made for new applications and functionalities to the fields of molecular biology, genetic analysis and proteomics, enabling the expansion of the cell biology field. Specifically, microfluidics has provided promising tools for enhancing cell biological research, since it has the ability to precisely control the cellular environment, to easily mimic heterogeneous cellular environment by multiplexing, and to analyze sub-cellular information by high-contents screening assays at the single-cell level. Various cell manipulation techniques in microfluidics have been developed in accordance with specific objectives and applications. In this review, we examine the latest achievements of cell manipulation techniques in microfluidics by categorizing externally applied forces for manipulation: (i) optical, (ii) magnetic, (iii) electrical, (iv) mechanical and (v) other manipulations. We furthermore focus on history where the manipulation techniques originate and also discuss future perspectives with key examples where available.

  4. 1 Hz rTMS of the left posterior parietal cortex (PPC) modifies sensorimotor timing

    PubMed Central

    Krause, Vanessa; Bashir, Shahid; Pollok, Bettina; Caipa, Anuhya; Schnitzler, Alfons; Pascual-Leone, Alvaro

    2013-01-01

    In order to investigate the relevance of the left posterior parietal cortex (PPC) for precise sensorimotor timing we applied 1 Hz repetitive transcranial magnetic stimulation (rTMS) over left PPC, right PPC and visual cortex of healthy participants for ten minutes, respectively. The impact on sensorimotor timing of the right hand was assessed using a synchronization task that required subjects to synchronize their right index finger taps with respect to constant auditory, visual or auditory-visual pacing. Our results reveal reduced negative tap-to-pacer asynchronies following rTMS of the left PPC in all pacing conditions. This effect lasted for about 5 minutes after cessation of rTMS. Right PPC and visual cortex stimulation did not yield any significant behavioural effects. Since suppression of left PPC modified right-hand synchronization accuracy independent of the pacing signal’s modality, the present data support the significance of left PPC for anticipatory motor control over a primary role in multisensory integration. The present data suggest that 1 Hz rTMS might interrupt a matching process of anticipated and real sensorimotor feedback within PPC. Alternatively, downregulation of left PPC activity may affect M1 excitability via functional connections leading to a delay in motor output and, thus, smaller tap-to-pacer asynchronies. PMID:23103789

  5. Droplet microfluidics based microseparation systems.

    PubMed

    Xiao, Zhiliang; Niu, Menglei; Zhang, Bo

    2012-06-01

    Lab on a chip (LOC) technology is a promising miniaturization approach. The feature that it significantly reduced sample consumption makes great sense in analytical and bioanalytical chemistry. Since the start of LOC technology, much attention has been focused on continuous flow microfluidic systems. At the turn of the century, droplet microfluidics, which was also termed segmented flow microfluidics, was introduced. Droplet microfluidics employs two immiscible phases to form discrete droplets, which are ideal vessels with confined volume, restricted dispersion, limited cross-contamination, and high surface area. Due to these unique features, droplet microfluidics proves to be a versatile tool in microscale sample handling. This article reviews the utility of droplet microfluidics in microanalytical systems with an emphasize on separation science, including sample encapsulation at ultra-small volume, compartmentalization of separation bands, isolation of droplet contents, and related detection techniques.

  6. Microfluidic redox battery.

    PubMed

    Lee, Jin Wook; Goulet, Marc-Antoni; Kjeang, Erik

    2013-07-07

    A miniaturized microfluidic battery is proposed, which is the first membraneless redox battery demonstrated to date. This unique concept capitalizes on dual-pass flow-through porous electrodes combined with stratified, co-laminar flow to generate electrical power on-chip. The fluidic design is symmetric to allow for both charging and discharging operations in forward, reverse, and recirculation modes. The proof-of-concept device fabricated using low-cost materials integrated in a microfluidic chip is shown to produce competitive power levels when operated on a vanadium redox electrolyte. A complete charge/discharge cycle is performed to demonstrate its operation as a rechargeable battery, which is an important step towards providing sustainable power to lab-on-a-chip and microelectronic applications.

  7. Ductular reaction-on-a-chip: Microfluidic co-cultures to study stem cell fate selection during liver injury

    PubMed Central

    Haque, Amranul; Gheibi, Pantea; Stybayeva, Gulnaz; Gao, Yandong; Torok, Natalie; Revzin, Alexander

    2016-01-01

    Liver injury modulates local microenvironment, triggering production of signals that instruct stem cell fate choices. In this study, we employed a microfluidic co-culture system to recreate important interactions in the liver stem cell niche, those between adult hepatocytes and liver progenitor cells (LPCs). We demonstrate that pluripotent stem cell-derived LPCs choose hepatic fate when cultured next to healthy hepatocytes but begin biliary differentiation program when co-cultured with injured hepatocytes. We connect this fate selection to skewing in production of hepatocyte growth factor (HGF) and transforming growth factor (TGF)-β1 caused by injury. Significantly, biliary fate selection of LPCs was not observed in the absence of hepatocytes nor did it happen in the presence of TGF-β inhibitors. Our study demonstrates that microfluidic culture systems may offer an interesting new tool for dissecting cellular interactions leading to aberrant stem cell differentiation during injury. PMID:27796316

  8. Microfluidic-chip-based multiple-locus variable-number tandem-repeat fingerprinting with new primer sets for methicillin-resistant Staphylococcus aureus.

    PubMed

    Sabat, Artur J; Chlebowicz, Monika A; Grundmann, Hajo; Arends, Jan P; Kampinga, Greetje; Meessen, Nico E L; Friedrich, Alexander W; van Dijl, Jan Maarten

    2012-07-01

    The detection of outbreaks of methicillin-resistant Staphylococcus aureus (MRSA) infections and a rapid and accurate identification of sources and routes of transmission should be conducted in hospital settings as early and swiftly as possible. In this study, we investigated the application potential of a new approach based on multiple-locus variable-number tandem-repeat fingerprinting (MLVF) and microfluidics technology for a rapid discrimination of MRSA lineages in outbreak settings. A total of 206 nonrepetitive MRSA isolates recovered from infected patients at the University Medical Center Groningen between 2000 and 2010 were tested. The results obtained by MLVF using microcapillary electrophoresis with newly designed primers were compared to those obtained by spa typing and multiple-locus variable-number tandem-repeat analysis (MLVA). The discriminatory power was 0.980 (107 patterns), 0.969 (85 allelic profiles), and 0.959 (66 types) for MLVF, MLVA, and spa typing, respectively. All methods tested showed a good concordance of results calculated by the adjusted Rand's coefficient method. Comparisons of data obtained by the three approaches allowed us to propose an 88% cutoff value for the similarity between any two MLVF patterns, which can be used in S. aureus epidemiological studies, including analyses of outbreaks and strain transmission events. Of the three tested methods, MLVF is the cheapest, fastest, and easiest to perform. MLVF applied to microfluidic polymer chips is a rapid, cheap, reproducible, and highly discriminating tool to determine the clonality of MRSA isolates and to trace the spread of MRSA strains over periods of many years. Although spa typing should be used due to its portability of data, MLVF has a high added value because it is more discriminatory.

  9. Punch Card Programmable Microfluidics

    PubMed Central

    Korir, George; Prakash, Manu

    2015-01-01

    Small volume fluid handling in single and multiphase microfluidics provides a promising strategy for efficient bio-chemical assays, low-cost point-of-care diagnostics and new approaches to scientific discoveries. However multiple barriers exist towards low-cost field deployment of programmable microfluidics. Incorporating multiple pumps, mixers and discrete valve based control of nanoliter fluids and droplets in an integrated, programmable manner without additional required external components has remained elusive. Combining the idea of punch card programming with arbitrary fluid control, here we describe a self-contained, hand-crank powered, multiplex and robust programmable microfluidic platform. A paper tape encodes information as a series of punched holes. A mechanical reader/actuator reads these paper tapes and correspondingly executes operations onto a microfluidic chip coupled to the platform in a plug-and-play fashion. Enabled by the complexity of codes that can be represented by a series of holes in punched paper tapes, we demonstrate independent control of 15 on-chip pumps with enhanced mixing, normally-closed valves and a novel on-demand impact-based droplet generator. We demonstrate robustness of operation by encoding a string of characters representing the word “PUNCHCARD MICROFLUIDICS” using the droplet generator. Multiplexing is demonstrated by implementing an example colorimetric water quality assays for pH, ammonia, nitrite and nitrate content in different water samples. With its portable and robust design, low cost and ease-of-use, we envision punch card programmable microfluidics will bring complex control of microfluidic chips into field-based applications in low-resource settings and in the hands of children around the world. PMID:25738834

  10. Semiconductor sensor embedded microfluidic chip for protein biomarker detection using a bead-based immunoassay combined with deoxyribonucleic acid strand labeling.

    PubMed

    Lin, Yen-Heng; Peng, Po-Yu

    2015-04-15

    Two major issues need to be addressed in applying semiconductor biosensors to detecting proteins in immunoassays. First, the length of the antibody on the sensor surface surpasses the Debye lengths (approximately 1 nm, in normal ionic strength solution), preventing certain specifically bound proteins from being tightly attached to the sensor surface. Therefore, these proteins do not contribute to the sensor's surface potential change. Second, these proteins carry a small charge and can be easily affected by the pH of the surrounding solution. This study proposes a magnetic bead-based immunoassay using a secondary antibody to label negatively charged DNA fragments for signal amplification. An externally imposed magnetic force attaches the analyte tightly to the sensor surface, thereby effectively solving the problem of the analyte protein's distance to the sensor surface surpassing the Debye lengths. In addition, a normal ion intensity buffer can be used without dilution for the proposed method. Experiments revealed that the sensitivity can be improved by using a longer DNA fragment for labeling and smaller magnetic beads as solid support for the antibody. By using a 90 base pair DNA label, the signal was 15 times greater than that without labeling. In addition, by using a 120 nm magnetic bead, a minimum detection limit of 12.5 ng mL(-1) apolipoprotein A1 can be measured. Furthermore, this study integrates a semiconductor sensor with a microfluidic chip. With the help of microvalves and micromixers in the chip, the length of the mixing step for each immunoassay has been reduced from 1h to 20 min, and the sample volume has been reduced from 80 μL to 10 μL. In practice, a protein biomarker in a urinary bladder cancer patient's urine was successfully measured using this technique. This study provides a convenient and effective method to measure protein using a semiconductor sensor.

  11. Weight Loss by Ppc-1, a Novel Small Molecule Mitochondrial Uncoupler Derived from Slime Mold

    PubMed Central

    Suzuki, Toshiyuki; Kikuchi, Haruhisa; Ogura, Masato; Homma, Miwako K.; Oshima, Yoshiteru; Homma, Yoshimi

    2015-01-01

    Mitochondria play a key role in diverse processes including ATP synthesis and apoptosis. Mitochondrial function can be studied using inhibitors of respiration, and new agents are valuable for discovering novel mechanisms involved in mitochondrial regulation. Here, we screened small molecules derived from slime molds and other microorganisms for their effects on mitochondrial oxygen consumption. We identified Ppc-1 as a novel molecule which stimulates oxygen consumption without adverse effects on ATP production. The kinetic behavior of Ppc-1 suggests its function as a mitochondrial uncoupler. Serial administration of Ppc-1 into mice suppressed weight gain with no abnormal effects on liver or kidney tissues, and no evidence of tumor formation. Serum fatty acid levels were significantly elevated in mice treated with Ppc-1, while body fat content remained low. After a single administration, Ppc-1 distributes into various tissues of individual animals at low levels. Ppc-1 stimulates adipocytes in culture to release fatty acids, which might explain the elevated serum fatty acids in Ppc-1-treated mice. The results suggest that Ppc-1 is a unique mitochondrial regulator which will be a valuable tool for mitochondrial research as well as the development of new drugs to treat obesity. PMID:25668511

  12. Bioanalysis in structured microfluidic systems.

    PubMed

    Ros, Alexandra; Hellmich, Wibke; Regtmeier, Jan; Duong, Thanh Tu; Anselmetti, Dario

    2006-07-01

    Microfluidic and lab-on-a-chip devices have attracted widespread interest in separation sciences and bioanalysis. Recent designs in microfluidic devices extend common separation concepts by exploiting new phenomena for molecular dynamics on a length scale of 10 mum and below, giving rise to novel manipulation tools and nonintuitive phenomena for microseparations. Here, we focus on three very recent developments for bioseparations based on tailored microfluidic systems: Single cell navigation, trapping and steering with subsequent on-chip lysis, protein separation and LIF detection (Section 3.1), then we report dielectrophoretic trapping and separation of large DNA fragments in structured microfluidic devices (Section 3.2). Finally, a paradoxial migration phenomenon based on thermal fluctuations, periodically arranged microchannels and a biased alternating current electric field is presented in Section 3.3.

  13. Generation of microgrooved silica nanotube membranes with sustained drug delivery and cell contact guidance ability by using a Teflon microfluidic chip

    PubMed Central

    Chen, Song; Shi, Xuetao; Chinnathambi, Shanmugavel; Wu, Hongkai; Hanagata, Nobutaka

    2013-01-01

    Silica nanotubes have been extensively applied in the biomedical field. However, very little attention has been paid to the fabrication and application of micropatterned silica nanotubes. In the present study, microgrooved silica nanotube membranes were fabricated in situ by microgrooving silica-coated collagen hybrid fibril hydrogels in a Teflon microfluidic chip followed by calcination for removal of collagen fibrils. Scanning electron microscopy images showed that the resulting silica nanotube membranes displayed a typical microgroove/ridge surface topography with ∼50 μm microgroove width and ∼120 μm ridge width. They supported adsorption of bone morphogenetic protein 2 (BMP-2) and exhibited a sustained release behavior for BMP-2. After culturing with osteoblast MC3T3-E1 cells, they induced an enhanced osteoblast differentiation due to the release of biologically active BMP-2 and a strong contact guidance ability to directly align and elongate osteoblasts due to the presence of microgrooved surface topography, indicating their potential application as a multi-functional cell-supporting matrix for tissue generation. PMID:27877563

  14. Kinsenoside screening with a microfluidic chip attenuates gouty arthritis through inactivating NF-κB signaling in macrophages and protecting endothelial cells

    PubMed Central

    Han, Qiao; Bing, Wang; Di, Yin; Hua, Li; Shi-he, Li; Yu-hua, Zheng; Xiu-guo, Han; Yu-gang, Wang; Qi-ming, Fan; Shih-mo, Yang; Ting-ting, Tang

    2016-01-01

    Gouty arthritis is a rheumatic disease that is characterized by the deposition of monosodium urate (MSU) in synovial joints cause by the increased serum hyperuricemia. This study used a three-dimensional (3D) flowing microfluidic chip to screen the effective candidate against MSU-stimulated human umbilical vein endothelial cell (HUVEC) damage, and found kinsenoside (Kin) to be the leading active component of Anoectochilus roxburghi, one of the Chinese medicinal plant widely used in the treatment of gouty arthritis clinically. Cell viability and apoptosis of HUVECs were evaluated, indicating that direct Kin stimulation and conditioned medium (CM) from Kin-treated macrophages both negatively modulated with MSU crystals. Additionally, Kin was capable of attenuating MSU-induced activation of nuclear factor-κB/mitogen-activated protein kinase (NF-κB/MAPK) signaling, targeting IκB kinase-α (IKKα) and IKKβ kinases of macrophages and influencing the expressions of NF-κB downstream cytokines and subsequent HUVEC bioactivity. Inflammasome NLR pyrin domain-containing 3 (NALP3) and toll-like receptor 2 (TLR2) were also inhibited after Kin treatment. Also, Kin downregulated CD14-mediated MSU crystals uptake in macrophages. In vivo study with MSU-injected ankle joints further revealed the significant suppression of inflammatory infiltration and endothelia impairment coupled with alleviation of ankle swelling and nociceptive response via Kin treatments. Taken together, these data implicated that Kin was the most effective candidate from Anoectochilus roxburghi to treat gouty arthritis clinically. PMID:27584788

  15. Generation of microgrooved silica nanotube membranes with sustained drug delivery and cell contact guidance ability by using a Teflon microfluidic chip

    NASA Astrophysics Data System (ADS)

    Chen, Song; Shi, Xuetao; Chinnathambi, Shanmugavel; Wu, Hongkai; Hanagata, Nobutaka

    2013-02-01

    Silica nanotubes have been extensively applied in the biomedical field. However, very little attention has been paid to the fabrication and application of micropatterned silica nanotubes. In the present study, microgrooved silica nanotube membranes were fabricated in situ by microgrooving silica-coated collagen hybrid fibril hydrogels in a Teflon microfluidic chip followed by calcination for removal of collagen fibrils. Scanning electron microscopy images showed that the resulting silica nanotube membranes displayed a typical microgroove/ridge surface topography with ˜50 μm microgroove width and ˜120 μm ridge width. They supported adsorption of bone morphogenetic protein 2 (BMP-2) and exhibited a sustained release behavior for BMP-2. After culturing with osteoblast MC3T3-E1 cells, they induced an enhanced osteoblast differentiation due to the release of biologically active BMP-2 and a strong contact guidance ability to directly align and elongate osteoblasts due to the presence of microgrooved surface topography, indicating their potential application as a multi-functional cell-supporting matrix for tissue generation.

  16. An Impedance Aptasensor with Microfluidic Chips for Specific Detection of H5N1 Avian Influenza Virus

    PubMed Central

    Lum, Jacob; Wang, Ronghui; Hargis, Billy; Tung, Steve; Bottje, Walter; Lu, Huaguang; Li, Yanbin

    2015-01-01

    In this research a DNA aptamer, which was selected through SELEX (systematic evolution of ligands by exponential enrichment) to be specific against the H5N1 subtype of the avian influenza virus (AIV), was used as an alternative reagent to monoclonal antibodies in an impedance biosensor utilizing a microfluidics flow cell and an interdigitated microelectrode for the specific detection of H5N1 AIV. The gold surface of the interdigitated microelectrode embedded in a microfluidics flow cell was modified using streptavidin. The biotinylated aptamer against H5N1 was then immobilized on the electrode surface using biotin–streptavidin binding. The target virus was captured on the microelectrode surface, causing an increase in impedance magnitude. The aptasensor had a detection time of 30 min with a detection limit of 0.0128 hemagglutinin units (HAU). Scanning electron microscopy confirmed the binding of the target virus onto the electrode surface. The DNA aptamer was specific to H5N1 and had no cross-reaction to other subtypes of AIV (e.g., H1N1, H2N2, H7N2). The newly developed aptasensor offers a portable, rapid, low-cost alternative to current methods with the same sensitivity and specificity. PMID:26230699

  17. Integrated fluorescence detection of labeled biomolecules using a prism-like PDMS microfluidic chip and lateral light excitation.

    PubMed

    Novo, Pedro; Chu, Virginia; Conde, João Pedro

    2014-06-21

    Microfabricated amorphous silicon photodiodes were integrated with prism-like PDMS microfluidics for the detection and quantification of fluorescence signals. The PDMS device was fabricated with optical quality surfaces and beveled sides. A 405 nm laser beam perpendicular to the lateral sides of the microfluidic device excites the fluorophores in the microchannel at an angle of 70° to the normal to the microchannel/photodiode surface. This configuration, which makes use of the total internal reflection of the excitation beam and the isotropy of the fluorescence emission, minimizes the intensity of excitation light that reaches the integrated photodetector. A difference of two orders of magnitude was achieved in the reduction of the detection noise level as compared with a normally incident excitation configuration. A limit-of-detection of 5.6 × 10(10) antibodies per square centimeter was achieved using antibodies labeled with a model organic fluorophore. Furthermore, the results using the lateral excitation scheme are in good proportionality agreement with those by fluorescence quantification using wide-field fluorescence microscopy.

  18. Surface acoustic wave microfluidics.

    PubMed

    Ding, Xiaoyun; Li, Peng; Lin, Sz-Chin Steven; Stratton, Zackary S; Nama, Nitesh; Guo, Feng; Slotcavage, Daniel; Mao, Xiaole; Shi, Jinjie; Costanzo, Francesco; Huang, Tony Jun

    2013-09-21

    The recent introduction of surface acoustic wave (SAW) technology onto lab-on-a-chip platforms has opened a new frontier in microfluidics. The advantages provided by such SAW microfluidics are numerous: simple fabrication, high biocompatibility, fast fluid actuation, versatility, compact and inexpensive devices and accessories, contact-free particle manipulation, and compatibility with other microfluidic components. We believe that these advantages enable SAW microfluidics to play a significant role in a variety of applications in biology, chemistry, engineering and medicine. In this review article, we discuss the theory underpinning SAWs and their interactions with particles and the contacting fluids in which they are suspended. We then review the SAW-enabled microfluidic devices demonstrated to date, starting with devices that accomplish fluid mixing and transport through the use of travelling SAW; we follow that by reviewing the more recent innovations achieved with standing SAW that enable such actions as particle/cell focusing, sorting and patterning. Finally, we look forward and appraise where the discipline of SAW microfluidics could go next.

  19. Surface acoustic wave microfluidics

    PubMed Central

    Ding, Xiaoyun; Li, Peng; Lin, Sz-Chin Steven; Stratton, Zackary S.; Nama, Nitesh; Guo, Feng; Slotcavage, Daniel; Mao, Xiaole; Shi, Jinjie; Costanzo, Francesco; Huang, Tony Jun

    2014-01-01

    The recent introduction of surface acoustic wave (SAW) technology onto lab-on-a-chip platforms has opened a new frontier in microfluidics. The advantages provided by such SAW microfluidics are numerous: simple fabrication, high biocompatibility, fast fluid actuation, versatility, compact and inexpensive devices and accessories, contact-free particle manipulation, and compatibility with other microfluidic components. We believe that these advantages enable SAW microfluidics to play a significant role in a variety of applications in biology, chemistry, engineering, and medicine. In this review article, we discuss the theory underpinning SAWs and their interactions with particles and the contacting fluids in which they are suspended. We then review the SAW-enabled microfluidic devices demonstrated to date, starting with devices that accomplish fluid mixing and transport through the use of travelling SAW; we follow that by reviewing the more recent innovations achieved with standing SAW that enable such actions as particle/cell focusing, sorting, and patterning. Finally, we look forward and appraise where the discipline of SAW microfluidics could go next. PMID:23900527

  20. Microfluidic microarray systems and methods thereof

    SciTech Connect

    West, Jay A. A.; Hukari, Kyle W.; Hux, Gary A.

    2009-04-28

    Disclosed are systems that include a manifold in fluid communication with a microfluidic chip having a microarray, an illuminator, and a detector in optical communication with the microarray. Methods for using these systems for biological detection are also disclosed.

  1. Microfluidic Chip for High Efficiency Electrophoretic Analysis of Segmented Flow from a Microdialysis Probe and in Vivo Chemical Monitoring

    PubMed Central

    Wang, Meng; Roman, Gregory T.; Perry, Maura L.; Kennedy, Robert T.

    2009-01-01

    An effective method for in vivo chemical monitoring is to couple sampling probes, such as microdialysis, to on-line analytical methods. A limitation of this approach is that in vivo chemical dynamics may be distorted by flow and diffusion broadening during transfer from sampling probe to analytical system. Converting a homogenous sample stream to segmented flow can prevent such broadening. We have developed a system for coupling segmented microdialysis flow with chip-based electrophoresis. In this system, the dialysis probe is integrated with a PDMS chip that merges dialysate with fluorogenic reagent and segments the flow into 8–10 nL plugs at 0.3–0.5 Hz separated by perfluorodecalin. The plugs flow to a glass chip where they are extracted to an aqueous stream and analyzed by electrophoresis with fluorescence detection. The novel extraction system connects the segmented flow to an electrophoresis sampling channel by a shallow and hydrophilic extraction bridge that removes the entire aqueous droplet from the oil stream. With this approach, temporal resolution was 35 s and independent of distance between sampling and analysis. Electrophoretic analysis produced separation with 223,000 ± 21,000 theoretical plates, 4.4% RSD in peak height, and detection limits of 90–180 nM for six amino acids. This performance was made possible by three key elements: 1) reliable transfer of plug flow to a glass chip; 2) efficient extraction of aqueous plugs from segmented flow; and 3) electrophoretic injection suitable for high efficiency separation with minimal dilution of sample. The system was used to detect rapid concentration changes evoked by infusing glutamate uptake inhibitor into the striatum of anesthetized rats. These results demonstrate the potential of incorporating segmented flow into separations-based sensing schemes for studying chemical dynamics in vivo with improved temporal resolution. PMID:19803495

  2. [Recent development of microfluidic diagnostic technologies].

    PubMed

    Li, Haifang; Zhang, Qianyun; Lin, Jin-Ming

    2011-04-01

    Microfluidic devices exhibit a great promising development in clinical diagnosis and disease screening due to their advantages of precise controlling of fluid flow, requirement of miniamount sample, rapid reaction speed and convenient integration. In this paper, the improvements of microfluidic diagnostic technologies in recent years are reviewed. The applications and developments of on-chip disease marker detection, microfluidic cell selection and cell drug metabolism, and diagnostic micro-devices are discussed.

  3. A microfluidic D-subminiature connector.

    PubMed

    Scott, Adina; Au, Anthony K; Vinckenbosch, Elise; Folch, Albert

    2013-06-07

    Standardized, affordable, user-friendly world-to-chip interfaces represent one of the major barriers to the adoption of microfluidics. We present a connector system for plug-and-play interfacing of microfluidic devices to multiple input and output lines. The male connectors are based on existing standardized housings from electronics that are inexpensive and widely available. The female connectors are fabricated using familiar replica molding techniques that can easily be adopted by microfluidic developers.

  4. Rapid prototyping of a microfluidics-based Venturi micropump imprinted on polymeric, postage-stamp-sized chips

    NASA Astrophysics Data System (ADS)

    Curtis, C.; Eshaque, B.; Badali, K.; Karanassios, V.

    2012-06-01

    Pumps are widely used in chemical analysis. For instance, they are used to help transport liquid samples from a beaker to an instrument, for example for sample introduction. Pumps can also used to evacuate chambers used for mass spectrometry. For miniaturized, portable analytical instruments, miniaturized pumps are ideally suited. In this paper, a micropump with no moving parts that relies on the Venturi effect has been rapidly prototyped by imprinting fluidic channels on inexpensive polymeric substrates. The micropump was first evaluated for potential vacuum applications (e.g., for portable mass spectrometers). Subsequently, it was evaluated for its ability to transfer liquids in microfluidic channels (for possible use as a sample delivery vehicle to an appropriate sample introduction system).

  5. Effects of microchannel geometry on preconcentration intensity in microfluidic chips with straight or convergent-divergent microchannels.

    PubMed

    Chen, Chia-Lin; Yang, Ruey-Jen

    2012-03-01

    Preconcentration microfluidic devices are fabricated incorporating straight or convergent-divergent microchannels and hydrogel or Nafion membranes. Sample preconcentration is achieved utilizing concentration-polarization effects. The effects of the microchannel geometry on the preconcentration intensity are systematically examined. It is shown that for the preconcentrator with the straight microchannel, the time required to achieve a satisfactory preconcentration intensity increases with an increasing channel depth. For the convergent-divergent microchannel, the preconcentration intensity increases with a reducing convergent channel width. Comparing the preconcentration performance of the two different microchannel configurations, it is found that for an equivalent width of the main microchannel, the concentration effect in the convergent-divergent microchannel is faster than that in the straight microchannel.

  6. Rapid wasted-free microfluidic fabrication based on ink-jet approach for microfluidic sensing applications

    NASA Astrophysics Data System (ADS)

    Jarujareet, Ungkarn; Amarit, Rattasart; Sumriddetchkajorn, Sarun

    2016-11-01

    Realizing that current microfluidic chip fabrication techniques are time consuming and labor intensive as well as always have material leftover after chip fabrication, this research work proposes an innovative approach for rapid microfluidic chip production. The key idea relies on a combination of a widely-used inkjet printing method and a heat-based polymer curing technique with an electronic-mechanical control, thus eliminating the need of masking and molds compared to typical microfluidic fabrication processes. In addition, as the appropriate amount of polymer is utilized during printing, there is much less amount of material wasted. Our inkjet-based microfluidic printer can print out the desired microfluidic chip pattern directly onto a heated glass surface, where the printed polymer is suddenly cured. Our proof-of-concept demonstration for widely-used single-flow channel, Y-junction, and T-junction microfluidic chips shows that the whole microfluidic chip fabrication process requires only 3 steps with a fabrication time of 6 minutes.

  7. Paper spray mass spectrometry-based method for analysis of droplets in a gravity-driven microfluidic chip.

    PubMed

    Zhang, Yandong; Li, Haifang; Ma, Yuan; Lin, Jin-Ming

    2014-03-07

    This work presents a paper spray mass spectrometry-based method, to analyze microdroplets produced in a gravity-driven microchip. Droplets at ambient pressure were passively transferred from the chip to a paper substrate by the capillary wicking effect. Paper spray ionization was then performed for mass spectrometry (MS) analysis of droplet contents. The qualitative and quantitative analytical performances of this technique for single droplets were demonstrated. This manually controlled interface is straightforward, low-cost and simple to implement. Moreover, paper spray ionization MS holds promise in the direct analysis of real biological/chemical microreaction samples because of its tolerance with complex matrices. As a proof-of-concept example, the droplet-based acetylcholine hydrolysis was carried out to demonstrate the validation of our method for the direct analysis of micro-chemical/biological reactions. We also introduced a flow injection analysis (FIA) system combined with our droplet system to generate a concentration gradient. As a result, the microreaction can be performed at different concentrations and kinetic information can be obtained in one sample injection. In conclusion, the combination of a microdroplet chip with paper spray ionization and the introduction of the FIA system and make our droplet-MS scheme a useful platform for monitoring and analyzing organic-phase chemical/biological reactions.

  8. Heteronuclear Micro-Helmholtz Coil Facilitates µm-Range Spatial and Sub-Hz Spectral Resolution NMR of nL-Volume Samples on Customisable Microfluidic Chips.

    PubMed

    Spengler, Nils; Höfflin, Jens; Moazenzadeh, Ali; Mager, Dario; MacKinnon, Neil; Badilita, Vlad; Wallrabe, Ulrike; Korvink, Jan G

    2016-01-01

    We present a completely revised generation of a modular micro-NMR detector, featuring an active sample volume of ∼ 100 nL, and an improvement of 87% in probe efficiency. The detector is capable of rapidly screening different samples using exchangeable, application-specific, MEMS-fabricated, microfluidic sample containers. In contrast to our previous design, the sample holder chips can be simply sealed with adhesive tape, with excellent adhesion due to the smooth surfaces surrounding the fluidic ports, and so withstand pressures of ∼2.5 bar, while simultaneously enabling high spectral resolution up to 0.62 Hz for H2O, due to its optimised geometry. We have additionally reworked the coil design and fabrication processes, replacing liquid photoresists by dry film stock, whose final thickness does not depend on accurate volume dispensing or precise levelling during curing. We further introduced mechanical alignment structures to avoid time-intensive optical alignment of the chip stacks during assembly, while we exchanged the laser-cut, PMMA spacers by diced glass spacers, which are not susceptible to melting during cutting. Doing so led to an overall simplification of the entire fabrication chain, while simultaneously increasing the yield, due to an improved uniformity of thickness of the individual layers, and in addition, due to more accurate vertical positioning of the wirebonded coils, now delimited by a post base plateau. We demonstrate the capability of the design by acquiring a 1H spectrum of ∼ 11 nmol sucrose dissolved in D2O, where we achieved a linewidth of 1.25 Hz for the TSP reference peak. Chemical shift imaging experiments were further recorded from voxel volumes of only ∼ 1.5 nL, which corresponded to amounts of just 1.5 nmol per voxel for a 1 M concentration. To extend the micro-detector to other nuclei of interest, we have implemented a trap circuit, enabling heteronuclear spectroscopy, demonstrated by two 1H/13C 2D HSQC experiments.

  9. Heteronuclear Micro-Helmholtz Coil Facilitates µm-Range Spatial and Sub-Hz Spectral Resolution NMR of nL-Volume Samples on Customisable Microfluidic Chips

    PubMed Central

    Spengler, Nils; Höfflin, Jens; Moazenzadeh, Ali; Mager, Dario; MacKinnon, Neil; Badilita, Vlad; Wallrabe, Ulrike; Korvink, Jan G.

    2016-01-01

    We present a completely revised generation of a modular micro-NMR detector, featuring an active sample volume of ∼ 100 nL, and an improvement of 87% in probe efficiency. The detector is capable of rapidly screening different samples using exchangeable, application-specific, MEMS-fabricated, microfluidic sample containers. In contrast to our previous design, the sample holder chips can be simply sealed with adhesive tape, with excellent adhesion due to the smooth surfaces surrounding the fluidic ports, and so withstand pressures of ∼2.5 bar, while simultaneously enabling high spectral resolution up to 0.62 Hz for H2O, due to its optimised geometry. We have additionally reworked the coil design and fabrication processes, replacing liquid photoresists by dry film stock, whose final thickness does not depend on accurate volume dispensing or precise levelling during curing. We further introduced mechanical alignment structures to avoid time-intensive optical alignment of the chip stacks during assembly, while we exchanged the laser-cut, PMMA spacers by diced glass spacers, which are not susceptible to melting during cutting. Doing so led to an overall simplification of the entire fabrication chain, while simultaneously increasing the yield, due to an improved uniformity of thickness of the individual layers, and in addition, due to more accurate vertical positioning of the wirebonded coils, now delimited by a post base plateau. We demonstrate the capability of the design by acquiring a 1H spectrum of ∼ 11 nmol sucrose dissolved in D2O, where we achieved a linewidth of 1.25 Hz for the TSP reference peak. Chemical shift imaging experiments were further recorded from voxel volumes of only ∼ 1.5nL, which corresponded to amounts of just 1.5 nmol per voxel for a 1 M concentration. To extend the micro-detector to other nuclei of interest, we have implemented a trap circuit, enabling heteronuclear spectroscopy, demonstrated by two 1H/13C 2D HSQC experiments. PMID

  10. A Microfluidics-HPLC/Differential Mobility Spectrometer Macromolecular Detection System for Human and Robotic Missions

    NASA Technical Reports Server (NTRS)

    Coy, S. L.; Killeen, K.; Han, J.; Eiceman, G. A.; Kanik, I.; Kidd, R. D.

    2011-01-01

    Our goal is to develop a unique, miniaturized, solute analyzer based on microfluidics technology. The analyzer consists of an integrated microfluidics High Performance Liquid Chromatographic chip / Differential Mobility Spectrometer (?HPLCchip/ DMS) detection system

  11. Microfluidic devices with thick-film electrochemical detection

    DOEpatents

    Wang, Joseph; Tian, Baomin; Sahlin, Eskil

    2005-04-12

    An apparatus for conducting a microfluidic process and analysis, including at least one elongated microfluidic channel, fluidic transport means for transport of fluids through the microfluidic channel, and at least one thick-film electrode in fluidic connection with the outlet end of the microfluidic channel. The present invention includes an integrated on-chip combination reaction, separation and thick-film electrochemical detection microsystem, for use in detection of a wide range of analytes, and methods for the use thereof.

  12. 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.

  13. Development of a fast thermal response microfluidic system using liquid metal

    NASA Astrophysics Data System (ADS)

    Gao, Meng; Gui, Lin

    2016-07-01

    Room temperature liquid metal gallium alloy has been widely used in many micro-electromechanical systems applications, such as on-chip electrical microheaters, micro temperature sensors, micro pumps and so on. Injecting liquid metal into microchannels can provide a simple, rapid, low-cost but efficient way to integrate these elements in microfluidic chips with high accuracy. The liquid metal-filled microstructures can be designed in any shape and easily integrated into microfluidic chips. In this paper, an on-chip liquid metal-based thermal microfluidic system is proposed for quick temperature control at the microscale. The micro system utilizes just one microfluidic chip as a basic working platform, which has liquid metal-based on-chip heaters, temperature sensors and electroosmotic flow pumps. Under the comprehensive control of these elements, the micro system can quickly change the temperature of a target fluid in the microfluidic chip. These liquid metal-based on-chip elements are very helpful for the fabrication and miniaturization of the microfluidic chip. In this paper, deionized water is used to test the temperature control performance of the thermal microfluidic system. According to the experimental results, the micro system can efficiently control the temperature of water ranging from 28 °C to 90 °C. The thermal microfluidic system has great potential for use in many microfluidic applications, such as on-chip polymerase chain reaction, temperature gradient focusing, protein crystallization and chemical synthesis.

  14. On-chip sample preparation and analyte quantification using a microfluidic aqueous two-phase extraction coupled with an immunoassay.

    PubMed

    Soares, R R G; Novo, P; Azevedo, A M; Fernandes, P; Aires-Barros, M R; Chu, V; Conde, J P

    2014-11-07

    Immunoassays are fast and sensitive techniques for analyte quantification, and their use in point-of-care devices for medical, environmental, and food safety applications has potential benefits of cost, portability, and multiplexing. However, immunoassays are often affected by matrix interference effects, requiring the use of complex laboratory extraction and concentration procedures in order to achieve the required sensitivity. In this paper we propose an integrated microfluidic device for the simultaneous matrix clean-up, concentration and detection. This device consists of two modules in series, the first performing an aqueous two-phase extraction (ATPE) for matrix extraction and analyte pre-concentration, and the second an immunoassay for quantification. The model analyte was the mycotoxin ochratoxin A (OTA) in a wine matrix. Using this strategy, a limit of detection (LoD) of 0.26 ng mL(-1) was obtained for red wine spiked with OTA, well below the regulatory limit for OTA in wines of 2 ng mL(-1) set by the European Union. Furthermore, the linear response on the logarithmic concentration scale was observed to span 3 orders of magnitude (0.1-100 ng mL(-1)). These results are comparable to those obtained for the quantification of OTA in plain buffer without an integrated ATPE (LoD = 0.15 ng mL(-1)). The proposed method was also found to provide similar results for markedly different matrices, such as red and white wines. This novel approach based on aqueous two-phase systems can help the development of point-of-care devices that can directly deal with real samples in complex matrices without the need for extra extraction processes and equipment.

  15. Next generation MUT-MAP, a high-sensitivity high-throughput microfluidics chip-based mutation analysis panel.

    PubMed

    Schleifman, Erica B; Tam, Rachel; Patel, Rajesh; Tsan, Alison; Sumiyoshi, Teiko; Fu, Ling; Desai, Rupal; Schoenbrunner, Nancy; Myers, Thomas W; Bauer, Keith; Smith, Edward; Raja, Rajiv

    2014-01-01

    Molecular profiling of tumor tissue to detect alterations, such as oncogenic mutations, plays a vital role in determining treatment options in oncology. Hence, there is an increasing need for a robust and high-throughput technology to detect oncogenic hotspot mutations. Although commercial assays are available to detect genetic alterations in single genes, only a limited amount of tissue is often available from patients, requiring multiplexing to allow for simultaneous detection of mutations in many genes using low DNA input. Even though next-generation sequencing (NGS) platforms provide powerful tools for this purpose, they face challenges such as high cost, large DNA input requirement, complex data analysis, and long turnaround times, limiting their use in clinical settings. We report the development of the next generation mutation multi-analyte panel (MUT-MAP), a high-throughput microfluidic, panel for detecting 120 somatic mutations across eleven genes of therapeutic interest (AKT1, BRAF, EGFR, FGFR3, FLT3, HRAS, KIT, KRAS, MET, NRAS, and PIK3CA) using allele-specific PCR (AS-PCR) and Taqman technology. This mutation panel requires as little as 2 ng of high quality DNA from fresh frozen or 100 ng of DNA from formalin-fixed paraffin-embedded (FFPE) tissues. Mutation calls, including an automated data analysis process, have been implemented to run 88 samples per day. Validation of this platform using plasmids showed robust signal and low cross-reactivity in all of the newly added assays and mutation calls in cell line samples were found to be consistent with the Catalogue of Somatic Mutations in Cancer (COSMIC) database allowing for direct comparison of our platform to Sanger sequencing. High correlation with NGS when compared to the SuraSeq500 panel run on the Ion Torrent platform in a FFPE dilution experiment showed assay sensitivity down to 0.45%. This multiplexed mutation panel is a valuable tool for high-throughput biomarker discovery in personalized

  16. New materials for microfluidics in biology.

    PubMed

    Ren, Kangning; Chen, Yin; Wu, Hongkai

    2014-02-01

    With its continuous progress, microfluidics has become a key enabling technology in biological research. During the past few years, the major growth of microfluidics shifted to the introduction of new materials in making microfluidic chips, primarily driven by the demand of versatile strategies to interface microfluidics with biological cell studies. Although polydimethylsiloxane is still used as primary frame material, hydrogels have been increasingly employed in cell-culture related applications. Moreover, plastics and paper are attracting more attention in commercial device fabrication. Aiming to reflect this trend, current review focuses on the progress of microfluidic chip materials over the time span of January 2011 through June 2013, and provides critical discussion of the resulting major new tools in biological research.

  17. Orientation-Based Control of Microfluidics

    PubMed Central

    Norouzi, Nazila; Bhakta, Heran C.; Grover, William H.

    2016-01-01

    Most microfluidic chips utilize off-chip hardware (syringe pumps, computer-controlled solenoid valves, pressure regulators, etc.) to control fluid flow on-chip. This expensive, bulky, and power-consuming hardware severely limits the utility of microfluidic instruments in resource-limited or point-of-care contexts, where the cost, size, and power consumption of the instrument must be limited. In this work, we present a technique for on-chip fluid control that requires no off-chip hardware. We accomplish this by using inert compounds to change the density of one fluid in the chip. If one fluid is made 2% more dense than a second fluid, when the fluids flow together under laminar flow the interface between the fluids quickly reorients to be orthogonal to Earth’s gravitational force. If the channel containing the fluids then splits into two channels, the amount of each fluid flowing into each channel is precisely determined by the angle of the channels relative to gravity. Thus, any fluid can be routed in any direction and mixed in any desired ratio on-chip simply by holding the chip at a certain angle. This approach allows for sophisticated control of on-chip fluids with no off-chip control hardware, significantly reducing the cost of microfluidic instruments in point-of-care or resource-limited settings. PMID:26950700

  18. Oxygen control with microfluidics.

    PubMed

    Brennan, Martin D; Rexius-Hall, Megan L; Elgass, Laura Jane; Eddington, David T

    2014-11-21

    Cellular function and behavior are affected by the partial pressure of O2, or oxygen tension, in the microenvironment. The level of oxygenation is important, as it is a balance of oxygen availability and oxygen consumption that is necessary to maintain normoxia. Changes in oxygen tension, from above physiological oxygen tension (hyperoxia) to below physiological levels (hypoxia) or even complete absence of oxygen (anoxia), trigger potent biological responses. For instance, hypoxia has been shown to support the maintenance and promote proliferation of regenerative stem and progenitor cells. Paradoxically, hypoxia also contributes to the development of pathological conditions including systemic inflammatory response, tumorigenesis, and cardiovascular disease, such as ischemic heart disease and pulmonary hypertension. Current methods to study cellular behavior in low levels of oxygen tension include hypoxia workstations and hypoxia chambers. These culture systems do not provide oxygen gradients that are found in vivo or precise control at the microscale. Microfluidic platforms have been developed to overcome the inherent limits of these current methods, including lack of spatial control, slow equilibration, and unachievable or difficult coupling to live-cell microscopy. The various applications made possible by microfluidic systems are the topic of this review. In order to understand how the microscale can be leveraged for oxygen control of cells and tissues within microfluidic systems, some background understanding of diffusion, solubility, and transport at the microscale will be presented in addition to a discussion on the methods for measuring the oxygen tension in microfluidic channels. Finally the various methods for oxygen control within microfluidic platforms will be discussed including devices that rely on diffusion from liquid or gas, utilizing on-or-off-chip mixers, leveraging cellular oxygen uptake to deplete the oxygen, relying on chemical reactions in

  19. Suspended microfluidics.

    PubMed

    Casavant, Benjamin P; Berthier, Erwin; Theberge, Ashleigh B; Berthier, Jean; Montanez-Sauri, Sara I; Bischel, Lauren L; Brakke, Kenneth; Hedman, Curtis J; Bushman, Wade; Keller, Nancy P; Beebe, David J

    2013-06-18

    Although the field of microfluidics has made significant progress in bringing new tools to address biological questions, the accessibility and adoption of microfluidics within the life sciences are still limited. Open microfluidic systems have the potential to lower the barriers to adoption, but the absence of robust design rules has hindered their use. Here, we present an open microfluidic platform, suspended microfluidics, that uses surface tension to fill and maintain a fluid in microscale structures devoid of a ceiling and floor. We developed a simple and ubiquitous model predicting fluid flow in suspended microfluidic systems and show that it encompasses many known capillary phenomena. Suspended microfluidics was used to create arrays of collagen membranes, mico Dots (μDots), in a horizontal plane separating two fluidic chambers, demonstrating a transwell platform able to discern collective or individual cellular invasion. Further, we demonstrated that μDots can also be used as a simple multiplexed 3D cellular growth platform. Using the μDot array, we probed the combined effects of soluble factors and matrix components, finding that laminin mitigates the growth suppression properties of the matrix metalloproteinase inhibitor GM6001. Based on the same fluidic principles, we created a suspended microfluidic metabolite extraction platform using a multilayer biphasic system that leverages the accessibility of open microchannels to retrieve steroids and other metabolites readily from cell culture. Suspended microfluidics brings the high degree of fluidic control and unique functionality of closed microfluidics into the highly accessible and robust platform of open microfluidics.

  20. On-demand microfluidic droplet manipulation using hydrophobic ferrofluid as a continuous-phase.

    PubMed

    Zhang, Kai; Liang, Qionglin; Ai, Xiaoni; Hu, Ping; Wang, Yiming; Luo, Guoan

    2011-04-07

    Multiple essential microdroplet operation units, including splitting, dispensing, oil-phase exchange, trapping, release and demulsification, were successfully implemented by combining hydrophobic ferrofluid with microfluidic chips.

  1. Design and Characterization of a Sensorized Microfluidic Cell-Culture System with Electro-Thermal Micro-Pumps and Sensors for Cell Adhesion, Oxygen, and pH on a Glass Chip

    PubMed Central

    Bonk, Sebastian M.; Stubbe, Marco; Buehler, Sebastian M.; Tautorat, Carsten; Baumann, Werner; Klinkenberg, Ernst-Dieter; Gimsa, Jan

    2015-01-01

    We combined a multi-sensor glass-chip with a microfluidic channel grid for the characterization of cellular behavior. The grid was imprinted in poly-dimethyl-siloxane. Mouse-embryonal/fetal calvaria fibroblasts (MC3T3-E1) were used as a model system. Thin-film platinum (Pt) sensors for respiration (amperometric oxygen electrode), acidification (potentiometric pH electrodes) and cell adhesion (interdigitated-electrodes structures, IDES) allowed us to monitor cell-physiological parameters as well as the cell-spreading behavior. Two on-chip electro-thermal micro-pumps (ETμPs) permitted the induction of medium flow in the system, e.g., for medium mixing and drug delivery. The glass-wafer technology ensured the microscopic observability of the on-chip cell culture. Connecting Pt structures were passivated by a 1.2 μm layer of silicon nitride (Si3N4). Thin Si3N4 layers (20 nm or 60 nm) were used as the sensitive material of the pH electrodes. These electrodes showed a linear behavior in the pH range from 4 to 9, with a sensitivity of up to 39 mV per pH step. The oxygen sensors were circular Pt electrodes with a sensor area of 78.5 μm2. Their sensitivity was 100 pA per 1% oxygen increase in the range from 0% to 21% oxygen (air saturated). Two different IDES geometries with 30- and 50-μm finger spacings showed comparable sensitivities in detecting the proliferation rate of MC3T3 cells. These cells were cultured for 11 days in vitro to test the biocompatibility, microfluidics and electric sensors of our system under standard laboratory conditions. PMID:26263849

  2. Active, Universal Particle Micromanipulators: CPUs for Microfluidics

    NASA Astrophysics Data System (ADS)

    Mezic, Igor; Bottausci, Frederic

    2007-11-01

    Current designs for Lab-on-a-Chip applications consist of a variety of separate microfluidic chambers and channels for functions such as concentration, separation, reaction and mixing of bioparticles in liquids. Here we advance an alternative concept, named μfCPU, the Microfluidic Central Processing Unit, where the key microfluidic operations are performed within a single enclosure, using software-based inputs rather than physical hardware changes, thus emulating the role of the Central Processing Unit in computers and cells in living organisms. We present an experimental embodiment of such a device and describe a variety of microfluidic manipulation tasks achieved in it by the use of a suite of electromotive and fluidic forces in a time-dependent way to produce on-demand functionality. We also discuss a new microfluidic devices architecture that utilizes μfCPU as the basic processing unit and uses centralized pumping instead of integrated microfluidic pumps.

  3. Microfluidic Approaches for Protein Crystal Structure Analysis.

    PubMed

    Maeki, Masatoshi; Yamaguchi, Hiroshi; Tokeshi, Manabu; Miyazaki, Masaya

    2016-01-01

    This review summarizes two microfluidic-based protein crystallization methods, protein crystallization behavior in the microfluidic devices, and their applications for X-ray crystal structure analysis. Microfluidic devices provide many advantages for protein crystallography; they require small sample volumes, provide high-throughput screening, and allow control of the protein crystallization. A droplet-based protein crystallization method is a useful technique for high-throughput screening and the formation of a single crystal without any complicated device fabrication process. Well-based microfluidic platforms also enable effective protein crystallization. This review also summarizes the protein crystal growth behavior in microfluidic devices as, is known from viewpoints of theoretical and experimental approaches. Finally, we introduce applications of microfluidic devices for on-chip crystal structure analysis.

  4. Microfluidic-Based Robotic Sampling System for Radioactive Solutions

    SciTech Connect

    Jack D. Law; Julia L. Tripp; Tara E. Smith; Veronica J. Rutledge; Troy G. Garn; John Svoboda; Larry Macaluso

    2014-02-01

    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 system and identified system modifications to optimize performance.

  5. Copper nanowires immobilized on the boards of microfluidic chips for the rapid and simultaneous diagnosis of galactosemia diseases in newborn urine samples.

    PubMed

    García, Miguel; Alonso-Fernández, José Ramón; Escarpa, Alberto

    2013-10-01

    Galactosemia is a rare disease that is diagnosed through the identification of different metabolite profiles. Therefore, the specific detection of galactose 1-phosphate (Gal 1-P), galactose (Gal), and uridyl diphosphate galactose (UDP-Gal) confirms type I, II, and III galactosemia diseases. Because of the low prevalence of galactosemia, sample availability is very scarce and screening methods to diagnose the illness are not commonly employed around the world. This work describes the coupling of microfluidic chips (MCs) to copper nanowires (CuNWs) as electrochemical detectors for the fast diagnosis of galactosemia in precious newborn urine samples. Conceptually speaking, we hypothesize that the inherent selectivity and sensitivity of CuNWs, toward galactosemia metabolites detection in connection with MC selectivity could allow the fast and simultaneous detection of the three galactosemia biomarkers, which implies the fast diagnosis of any galactosemia type in just one single analysis. Electrosynthesized CuNWs show a well-defined shape, with an average length of 6 μm and a width of 300 nm. The modified electrodes exhibited an enhanced electroactive surface area twice as high as the nonmodified ones. Very good intraelectrode repeatability with relative standard deviations (RSDs) of <8% (n = 10) and interelectrode reproducibility with RSDs of <12% (n = 5) were obtained, indicating an excellent stability of the nanoscaled electrochemical detector. Under optimum chemical (3 mM NaOH, pH 11.5), electrokinetic (separation voltage +750 V, injection +1500 V for 5 s) and electrochemical (E = +0.70 V in 3 mM NaOH, pH 11.5) conditions, galactosemia diseases were unequivocally identified, differentiating between type I, II, and III, using selected precious ill diagnosed newborn urine samples. Detection proceeded within less than 350 s, required negligible urine sample consumption, and displayed impressive signal-to-noise characteristics (ranging from 14 to 80) and micromolar

  6. Microfluidic electronics.

    PubMed

    Cheng, Shi; Wu, Zhigang

    2012-08-21

    Microfluidics, a field that has been well-established for several decades, has seen extensive applications in the areas of biology, chemistry, and medicine. However, it might be very hard to imagine how such soft microfluidic devices would be used in other areas, such as electronics, in which stiff, solid metals, insulators, and semiconductors have previously dominated. Very recently, things have radically changed. Taking advantage of native properties of microfluidics, advances in microfluidics-based electronics have shown great potential in numerous new appealing applications, e.g. bio-inspired devices, body-worn healthcare and medical sensing systems, and ergonomic units, in which conventional rigid, bulky electronics are facing insurmountable obstacles to fulfil the demand on comfortable user experience. Not only would the birth of microfluidic electronics contribute to both the microfluidics and electronics fields, but it may also shape the future of our daily life. Nevertheless, microfluidic electronics are still at a very early stage, and significant efforts in research and development are needed to advance this emerging field. The intention of this article is to review recent research outcomes in the field of microfluidic electronics, and address current technical challenges and issues. The outlook of future development in microfluidic electronic devices and systems, as well as new fabrication techniques, is also discussed. Moreover, the authors would like to inspire both the microfluidics and electronics communities to further exploit this newly-established field.

  7. Note: A portable Raman analyzer for microfluidic chips based on a dichroic beam splitter for integration of imaging and signal collection light paths

    SciTech Connect

    Geng, Yijia; Xu, Shuping; Xu, Weiqing; Chen, Lei; Chen, Gang; Bi, Wenbin; Cui, Haining

    2015-05-15

    An integrated and portable Raman analyzer featuring an inverted probe fixed on a motor-driving adjustable optical module was designed for the combination of a microfluidic system. It possesses a micro-imaging function. The inverted configuration is advantageous to locate and focus microfluidic channels. Different from commercial micro-imaging Raman spectrometers using manual switchable light path, this analyzer adopts a dichroic beam splitter for both imaging and signal collection light paths, which avoids movable parts and improves the integration and stability of optics. Combined with surface-enhanced Raman scattering technique, this portable Raman micro-analyzer is promising as a powerful tool for microfluidic analytics.

  8. Development of a multi-layer microfluidic array chip to culture and replate uniform-sized embryoid bodies without manual cell retrieval.

    PubMed

    Kang, Edward; Choi, Yoon Young; Jun, Yesl; Chung, Bong Geun; Lee, Sang-Hoon

    2010-10-21

    We have developed a multi-layer, microfluidic array platform containing concave microwells and flat cell culture chambers to culture embryonic stem (ES) cells and regulate uniform-sized embryoid body (EB) formation. The main advantage of this platform was that EBs cultured within the concave microwells of a bottom layer were automatically replated into flat cell culture chambers of a top layer, following inversion of the multi-layer microfluidic array platform. This allowed EB formation and EB replating to be controlled simultaneously inside a single microfluidic device without pipette-based manual cell retrieval, a drawback of previous EB culture methods.

  9. Carbon dioxide-based copolymers: environmental benefits of PPC, an industrially viable catalyst.

    PubMed

    Qin, Yusheng; Wang, Xianhong

    2010-11-01

    Carbon dioxide-based copolymers utilize the green house gas CO(2) and can be applied in research and industry. Here we focus on industrially viable CO(2)-based catalysts in China and beyond. Poly(propylene carbonate) (PPC), an alternating copolymer of CO(2) and propylene oxide, is one of the emerging low-cost biodegradable plastics. We describe the thermal and mechanical performances of as-polymerized PPC, where amorphous state, low glass transition temperature, and biodegradability are the three main properties. We also describe modification of the PPC, the so-called toughening and strengthening at high temperature, and plasticizing at low temperature, including incorporation of a third monomer unit by chemical terpolymerization, and introduction of special intermolecular interactions or crystallizable components by physical blending. The fast development in catalyst design and performance improvement for PPC has created new chances for industry. In particular, high molecular weight PPC from rare earth ternary catalyst is becoming an economically viable biodegradable plastic with tens of thousands of tons produced per year, providing a new solution to overcome the problem of high cost in biodegradable plastics.

  10. Next-generation integrated microfluidic circuits.

    PubMed

    Mosadegh, Bobak; Bersano-Begey, Tommaso; Park, Joong Yull; Burns, Mark A; Takayama, Shuichi

    2011-09-07

    This mini-review provides a brief overview of recent devices that use networks of elastomeric valves to minimize or eliminate the need for interconnections between microfluidic chips and external instruction lines that send flow control signals. Conventional microfluidic control mechanisms convey instruction signals in a parallel manner such that the number of instruction lines must increase as the number of independently operated valves increases. The devices described here circumvent this "tyranny of microfluidic interconnects" by the serial encoding of information to enable instruction of an arbitrary number of independent valves with a set number of control lines, or by the microfluidic circuit-embedded encoding of instructions to eliminate control lines altogether. Because the parallel instruction chips are the most historical and straightforward to design, they are still the most commonly used approach today. As requirements for instruction complexity, chip-to-chip communication, and real-time on-chip feedback flow control arise, the next generation of integrated microfluidic circuits will need to incorporate these latest interconnect flow control approaches.

  11. Evidence for PPC1, a determinant of the pilei-pellis color of Agaricus bisporus fruitbodies.

    PubMed

    Callac, P; Moquet, F; Imbernon, M; Guedes-Lafargue, M R; Mamoun, M; Olivier, J M

    1998-03-01

    In the present study, we investigated the genetic basis of mushroom cap color. In first generation hybrids between a brown isolate and the white commercial hybrid U 1, the white trait was recessive. Color was determined using color meter technology in second generation hybrids obtained by crossing the homokaryotic progeny of a first generation hybrid with a homokaryon from U 1. Statistical analysis revealed a bimodal distribution describing two classes of white and not-white hybrids. We postulate that a recessive allele at a single locus (PPC1) encodes the white pilei-pellis color. Joint segregation analyses indicated that PPC1 was linked to the ADH (alcohol dehydrogenase) locus. Through the analysis of the heterokaryotic progeny of the first generation hybrid, a recombination model is proposed in which PPC1 is located between the centromere and the ADH locus.

  12. Analysis of trapped oscillation modes in magnetized PPC and its tunability for variable plasma parameters

    NASA Astrophysics Data System (ADS)

    Mittal, Tanvi; Yadav, Rana Pratap; Bora, Dhiraj

    2017-01-01

    In this study, analysis of magnetized plasma photonic crystal (PPC) using 1-D modeling has been presented where effect of extra-ordinary modes of plasma on Photonic bandgaps (PBGs) is highlighted. The PPC characteristic has been studied in range of 1-140 GHz for the different structural parameters, plasma frequency, angle of incidence, collision frequency and applied magnetic field. In the PPC, presence of static magnetic field yields the extra-ordinary mode in plasma that introduces trapped oscillations in the PBGs. This has been comprehensively analyzed and presented in detail. A procedure has been developed to identify the frequency range of the trapped oscillation in PBG. Study also explores that the trapped oscillations can be shifted at any other position in a prescribed frequency band by having suitable parameters like electronics concentration, hybrid frequency and applied magnetic field etc. Presented work can be utilized to avoid the trapped oscillation which yields non linearity in PBG.

  13. Efficacy and safety of PPC-5650 on experimental rectal pain in patients with irritable bowel syndrome.

    PubMed

    Nielsen, Lecia Møller; Olesen, Anne Estrup; Andresen, Trine; Simrén, Magnus; Törnblom, Hans; Drewes, Asbjørn Mohr

    2015-02-01

    PPC-5650 is a new pharmacological agent that can modulate acid-sensing ion channel activity, leading to a reduction in the pain signal under up-regulated conditions. The non-clinical programme for PPC-5650 supported a role for this novel agent in the treatment of pain in patients with irritable bowel syndrome (IBS). In patients with IBS, the aims of the study were: (1) to assess the efficacy of a single bolus of PPC-5650 locally applied in the rectum using multi-modal stimulations of the recto sigmoid and (2) to assess the safety profile of PPC-5650. The study was a randomized, double-blind, placebo-controlled, cross-over trial in patients with IBS, excluding females of child-bearing potential. The study consisted of a training visit, study visit 1 and 2 and a follow-up visit. Rectosigmoid electrical, thermal and mechanical stimulations were performed, pain perception was rated on a pain intensity scale and referred pain areas were assessed. All adverse events were registered. Twenty-five patients with IBS were enrolled and completed the study (9 women and 16 men; mean age 50.4 ± 12.7 years). No effects of the study drug were found on any of the rectal stimulations or for referred pain areas (all p > 0.05). No significant or clinically relevant treatment-related differences were seen for the laboratory safety variables or any other reported adverse event. In conclusion, in patients with IBS on rectal sensitivity to multi-modal stimulations, PPC-5650 did not produce efficacy relative to placebo. The overall safety and tolerability of PPC-5650 was acceptable.

  14. Motion in microfluidic ratchets.

    PubMed

    Caballero, D; Katuri, J; Samitier, J; Sánchez, S

    2016-11-15

    The ubiquitous random motion of mesoscopic active particles, such as cells, can be "rectified" or directed by embedding the particles in systems containing local and periodic asymmetric cues. Incorporated on lab-on-a-chip devices, these microratchet-like structures can be used to self-propel fluids, transport particles, and direct cell motion in the absence of external power sources. In this Focus article we discuss recent advances in the use of ratchet-like geometries in microfluidics which could open new avenues in biomedicine for applications in diagnosis, cancer biology, and bioengineering.

  15. Microfluidic serial dilution ladder.

    PubMed

    Ahrar, Siavash; Hwang, Michelle; Duncan, Philip N; Hui, Elliot E

    2014-01-07

    Serial dilution is a fundamental procedure that is common to a large number of laboratory protocols. Automation of serial dilution is thus a valuable component for lab-on-a-chip systems. While a handful of different microfluidic strategies for serial dilution have been reported, approaches based on continuous flow mixing inherently consume larger amounts of sample volume and chip real estate. We employ valve-driven circulatory mixing to address these issues and also introduce a novel device structure to store each stage of the dilution process. The dilution strategy is based on sequentially mixing the rungs of a ladder structure. We demonstrate a 7-stage series of 1 : 1 dilutions with R(2) equal to 0.995 in an active device area of 1 cm(2).

  16. Principles, Techniques, and Applications of Tissue Microfluidics

    NASA Technical Reports Server (NTRS)

    Wade, Lawrence A.; Kartalov, Emil P.; Shibata, Darryl; Taylor, Clive

    2011-01-01

    The principle of tissue microfluidics and its resultant techniques has been applied to cell analysis. Building microfluidics to suit a particular tissue sample would allow the rapid, reliable, inexpensive, highly parallelized, selective extraction of chosen regions of tissue for purposes of further biochemical analysis. Furthermore, the applicability of the techniques ranges beyond the described pathology application. For example, they would also allow the posing and successful answering of new sets of questions in many areas of fundamental research. The proposed integration of microfluidic techniques and tissue slice samples is called tissue microfluidics because it molds the microfluidic architectures in accordance with each particular structure of each specific tissue sample. Thus, microfluidics can be built around the tissues, following the tissue structure, or alternatively, the microfluidics can be adapted to the specific geometry of particular tissues. By contrast, the traditional approach is that microfluidic devices are structured in accordance with engineering considerations, while the biological components in applied devices are forced to comply with these engineering presets. The proposed principles represent a paradigm shift in microfluidic technology in three important ways: Microfluidic devices are to be directly integrated with, onto, or around tissue samples, in contrast to the conventional method of off-chip sample extraction followed by sample insertion in microfluidic devices. Architectural and operational principles of microfluidic devices are to be subordinated to suit specific tissue structure and needs, in contrast to the conventional method of building devices according to fluidic function alone and without regard to tissue structure. Sample acquisition from tissue is to be performed on-chip and is to be integrated with the diagnostic measurement within the same device, in contrast to the conventional method of off-chip sample prep and

  17. [Application of microfluidics in aquatic environmental pollution analysis].

    PubMed

    Wang, Hu; Wei, Jun-Feng; Zheng, Guo-Xia

    2014-04-01

    Recently, a new type of chip technology, microfluidics, has received global attention for its rapid analysis speed, low reagent consumption, small size and simple operation, etc. Based on a micro-channel network and supported by a Micro-Electro-Mechanic System (MEMS), this technology integrates all the functions of a laboratory into one small piece of chip, which is called "lab on the chip". This paper presented a brief introduction about microfluidics and its representative developments. Future prospects in the aspects of instrument miniaturization, system integration, chip materials, and detection techniques, as well as the implementation of microfluidics in aquatic environmental pollutant analysis were thoroughly discussed. Some problems faced now were put forward. With the rapid progress in the microfluidics, a universal low-cost microchip capable of high speed multi-channel detection and integrated with many kinds of detection methods would be the research focus in the future.

  18. 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.

  19. Manipulation of microfluidic droplets by electrorheological fluid.

    PubMed

    Zhang, Menying; Gong, Xiuqing; Wen, Weijia

    2009-09-01

    Microfluidics, especially droplet microfluidics, attracts more and more researchers from diverse fields, because it requires fewer materials and less time, produces less waste and has the potential of highly integrated and computer-controlled reaction processes for chemistry and biology. Electrorheological fluid, especially giant electrorheological fluid (GERF), which is considered as a kind of smart material, has been applied to the microfluidic systems to achieve active and precise control of fluid by electrical signal. In this review article, we will introduce recent results of microfluidic droplet manipulation, GERF and some pertinent achievements by introducing GERF into microfluidic system: digital generation, manipulation of "smart droplets" and droplet manipulation by GERF. Once it is combined with real-time detection, integrated chip with multiple functions can be realized.

  20. High-Voltage CMOS Controller for Microfluidics.

    PubMed

    Khorasani, M; Behnam, M; van den Berg, L; Backhouse, C J; Elliott, D G

    2009-04-01

    A high-voltage microfluidic controller designed using DALSA semiconductor's 0.8-mum low-voltage/high-voltage complementary metal-oxide semiconductor/double diffused metal-oxide semiconductor process is presented. The chip's four high-voltage output drivers can switch 300 V, and the dc-dc boost converter can generate up to 68 V using external passive components. This integrated circuit represents an advancement in microfluidic technology when used in conjunction with a charge coupling device (CCD)-based optical system and a glass microfluidic channel, enabling a portable and cost-efficient platform for genetic analysis.

  1. Microfluidic device for acoustic cell lysis

    SciTech Connect

    Branch, Darren W.; Cooley, Erika Jane; Smith, Gennifer Tanabe; James, Conrad D.; McClain, Jaime L.

    2015-08-04

    A microfluidic acoustic-based cell lysing device that can be integrated with on-chip nucleic acid extraction. Using a bulk acoustic wave (BAW) transducer array, acoustic waves can be coupled into microfluidic cartridges resulting in the lysis of cells contained therein by localized acoustic pressure. Cellular materials can then be extracted from the lysed cells. For example, nucleic acids can be extracted from the lysate using silica-based sol-gel filled microchannels, nucleic acid binding magnetic beads, or Nafion-coated electrodes. Integration of cell lysis and nucleic acid extraction on-chip enables a small, portable system that allows for rapid analysis in the field.

  2. 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.

  3. Microfluidic photonic integrated circuits

    NASA Astrophysics Data System (ADS)

    Cho, Sung Hwan; Godin, Jessica; Chen, Chun Hao; Tsai, Frank S.; Lo, Yu-Hwa

    2008-11-01

    We report on the development of an inexpensive, portable lab-on-a-chip flow cytometer system in which microfluidics, photonics, and acoustics are integrated together to work synergistically. The system relies on fluid-filled twodimensional on-chip photonic components such as lenses, apertures, and slab waveguides to allow for illumination laser beam shaping, light scattering and fluorescence signal detection. Both scattered and fluorescent lights are detected by photodetectors after being collected and guided by the on-chip optics components (e.g. lenses and waveguides). The detected light signal is imported and amplified in real time and triggers the piezoelectric actuator so that the targeted samples are directed into desired reservoir for subsequent advanced analysis. The real-time, closed-loop control system is developed with field-programmable-gate-array (FPGA) implementation. The system enables high-throughput (1- 10kHz operation), high reliability and low-powered (<1mW) fluorescence activated cell sorting (FACS) on a chip. The microfabricated flow cytometer can potentially be used as a portable, inexpensive point-of-care device in resource poor environments.

  4. Bio-microfluidics: biomaterials and biomimetic designs.

    PubMed

    Domachuk, Peter; Tsioris, Konstantinos; Omenetto, Fiorenzo G; Kaplan, David L

    2010-01-12

    Bio-microfluidics applies biomaterials and biologically inspired structural designs (biomimetics) to microfluidic devices. Microfluidics, the techniques for constraining fluids on the micrometer and sub-micrometer scale, offer applications ranging from lab-on-a-chip to optofluidics. Despite this wealth of applications, the design of typical microfluidic devices imparts relatively simple, laminar behavior on fluids and is realized using materials and techniques from silicon planar fabrication. On the other hand, highly complex microfluidic behavior is commonplace in nature, where fluids with nonlinear rheology flow through chaotic vasculature composed from a range of biopolymers. In this Review, the current state of bio-microfluidic materials, designs and applications are examined. Biopolymers enable bio-microfluidic devices with versatile functionalization chemistries, flexibility in fabrication, and biocompatibility in vitro and in vivo. Polymeric materials such as alginate, collagen, chitosan, and silk are being explored as bulk and film materials for bio-microfluidics. Hydrogels offer options for mechanically functional devices for microfluidic systems such as self-regulating valves, microlens arrays and drug release systems, vital for integrated bio-microfluidic devices. These devices including growth factor gradients to study cell responses, blood analysis, biomimetic capillary designs, and blood vessel tissue culture systems, as some recent examples of inroads in the field that should lead the way in a new generation of microfluidic devices for bio-related needs and applications. Perhaps one of the most intriguing directions for the future will be fully implantable microfluidic devices that will also integrate with existing vasculature and slowly degrade to fully recapitulate native tissue structure and function, yet serve critical interim functions, such as tissue maintenance, drug release, mechanical support, and cell delivery.

  5. QR-on-a-chip: a computer-recognizable micro-pattern engraved microfluidic device for high-throughput image acquisition.

    PubMed

    Yun, Kyungwon; Lee, Hyunjae; Bang, Hyunwoo; Jeon, Noo Li

    2016-02-21

    This study proposes a novel way to achieve high-throughput image acquisition based on a computer-recognizable micro-pattern implemented on a microfluidic device. We integrated the QR code, a two-dimensional barcode system, onto the microfluidic device to simplify imaging of multiple ROIs (regions of interest). A standard QR code pattern was modified to arrays of cylindrical structures of polydimethylsiloxane (PDMS). Utilizing the recognition of the micro-pattern, the proposed system enables: (1) device identification, which allows referencing additional information of the device, such as device imaging sequences or the ROIs and (2) composing a coordinate system for an arbitrarily located microfluidic device with respect to the stage. Based on these functionalities, the proposed method performs one-step high-throughput imaging for data acquisition in microfluidic devices without further manual exploration and locating of the desired ROIs. In our experience, the proposed method significantly reduced the time for the preparation of an acquisition. We expect that the method will innovatively improve the prototype device data acquisition and analysis.

  6. Microplasma fabrication: from semiconductor technology for 2D-chips and microfluidic channels to rapid prototyping and 3D-printing of microplasma devices

    NASA Astrophysics Data System (ADS)

    Shatford, R.; Karanassios, Vassili

    2014-05-01

    Microplasmas are receiving attention in recent conferences and current scientific literature. In our laboratory, microplasmas-on-chips proved to be particularly attractive. The 2D- and 3D-chips we developed became hybrid because they were fitted with a quartz plate (quartz was used due to its transparency to UV). Fabrication of 2D- and 3D-chips for microplasma research is described. The fabrication methods described ranged from semiconductor fabrication technology, to Computer Numerical Control (CNC) machining, to 3D-printing. These methods may prove to be useful for those contemplating in entering microplasma research but have no access to expensive semiconductor fabrication equipment.

  7. Identification and expression of a soybean nodule-enhanced PEP-carboxylase kinase gene (NE-PpcK) that shows striking up-/down-regulation in vivo.

    PubMed

    Xu, Wenxin; Zhou, You; Chollet, Raymond

    2003-05-01

    Various isoforms of plant phosphoenolpyruvate carboxylase (PEPC (Ppc)) are controlled post-translationally by an intricate interaction between allosteric regulation and reversible protein phosphorylation. In leaves and root nodules of legumes, these changes in PEPC phosphorylation state are governed primarily by PEPC-kinase (PpcK), a novel, 'minimal but functional' Ser/Thr kinase. To date, this plant-specific kinase has been investigated in molecular terms exclusively in non-leguminous plants, such as Crassulacean-acid-metabolism (CAM) species and Arabidopsis. As an important extension of our earlier biochemical studies on this dedicated kinase and PEPC phosphorylation in soybean (Glycine max) nodules, we now report the molecular cloning of the first legume PpcK from a soybean nodule cDNA library, which encodes a functional, 31.0 kDa PpcK polypeptide. Besides displaying organ, developmental, and spatial expression properties that are strikingly up-regulated in mature nodules, the expression pattern of this transcript is distinct from that of a second soybean PpcK isogene (GmPpcK). The steady-state abundance of this former, nodule-enhanced transcript (NE-PpcK) is markedly influenced by photosynthate supply from the shoots. This latter up-/down-regulation of NE-PpcK transcript level occurs in vivo in concert with the corresponding changes in the nodule PpcK activity, the phosphorylation-state of PEPC, and the abundance of a previously identified, nodule-enhanced transcript (GmPEPC7) that encodes the target enzyme (NE-Ppc). Furthermore, genomic Southern analysis and inspection of the public database indicate that there are at least three distinct PpcK and Ppc isogenes in soybean. Collectively, these and recent findings with Arabidopsis implicate the existence of multiple PpcK-Ppc'expression-partners' in plants, exemplified by NE-PpcK and NE-Ppc in the soybean nodule.

  8. Fabrication and characterization of polymer microfluidic devices for bio-agent detection

    NASA Astrophysics Data System (ADS)

    Morales, Alfredo M.; Brazzle, John D.; Crocker, Robert W.; Domeier, Linda A.; Goods, Eric B.; Hachman, John T., Jr.; Harnett, Cindy K.; Hunter, Marion C.; Mani, Seethambal S.; Mosier, Bruce P.; Simmons, Blake A.

    2004-12-01

    Sandia and Lawrence Livermore National Laboratories are developing a briefcase-sized, broad-spectrum bioagent detection system. This autonomous instrument, the BioBriefcase, will monitor the environment and warn against bacterium, virus, and toxin based biological attacks. At the heart of this device, inexpensive polymer microfluidic chips will carry out sample preparation and analysis. Fabrication of polymer microfluidic chips involves the creation of a master in etched glass; plating of the master to produce a nickel stamp; large lot chip replication by injection molding; and thermal chip sealing. Since the performance and reliability of microfluidic chips are very sensitive to fluidic impedance and to electromagnetic fluxes, the microchannel dimensions and shape have to be tightly controlled during chip fabrication. In this talk, we will present an overview of chip design and fabrication. Metrology data collected at different fabrication steps and the dimensional deviations of the polymer chip from the original design will be discussed.

  9. Fabrication and characterization of polymer microfluidic devices for bio-agent detection

    NASA Astrophysics Data System (ADS)

    Morales, Alfredo M.; Brazzle, John D.; Crocker, Robert W.; Domeier, Linda A.; Goods, Eric B.; Hachman, John T., Jr.; Harnett, Cindy K.; Hunter, Marion C.; Mani, Seethambal S.; Mosier, Bruce P.; Simmons, Blake A.

    2005-01-01

    Sandia and Lawrence Livermore National Laboratories are developing a briefcase-sized, broad-spectrum bioagent detection system. This autonomous instrument, the BioBriefcase, will monitor the environment and warn against bacterium, virus, and toxin based biological attacks. At the heart of this device, inexpensive polymer microfluidic chips will carry out sample preparation and analysis. Fabrication of polymer microfluidic chips involves the creation of a master in etched glass; plating of the master to produce a nickel stamp; large lot chip replication by injection molding; and thermal chip sealing. Since the performance and reliability of microfluidic chips are very sensitive to fluidic impedance and to electromagnetic fluxes, the microchannel dimensions and shape have to be tightly controlled during chip fabrication. In this talk, we will present an overview of chip design and fabrication. Metrology data collected at different fabrication steps and the dimensional deviations of the polymer chip from the original design will be discussed.

  10. Digital microfluidic operations on micro-electrode dot array architecture.

    PubMed

    Wang, G; Teng, D; Fan, S-K

    2011-12-01

    As digital microfluidics-based biochips find more applications, their complexity is expected to increase significantly owing to the trend of multiple and concurrent assays on the chip. There is a pressing need to deliver a top-down design methodology that the biochip designer can leverage the same level of computer-aided design support as the semi-conductor industry now does. Moreover, as microelectronics fabrication technology is scaling up and integrated device performance is improving, it is expected that these microfluidic biochips will be integrated with microelectronic components in next-generation system-on-chip designs. This study presents the analysis and experiments of digital microfluidic operations on a novel electrowetting-on-dielectric-based 'micro-electrode dot array architecture' that fosters a development path for hierarchical top-down design approach for digital microfluidics. The proposed architecture allows dynamic configurations and activations of identical basic microfluidic unit called 'micro-electrode cells' to design microfluidic components, layouts, routing, microfluidic operations and applications of the biochip hierarchically. Fundamental microfluidic operations have been successfully performed by the architecture. In addition, this novel architecture demonstrates a number of advantages and flexibilities over the conventional digital microfluidics in performing advanced microfluidic operations.

  11. Microfluidic Biochip Design

    NASA Technical Reports Server (NTRS)

    Panzarella, Charles

    2004-01-01

    As humans prepare for the exploration of our solar system, there is a growing need for miniaturized medical and environmental diagnostic devices for use on spacecrafts, especially during long-duration space missions where size and power requirements are critical. In recent years, the biochip (or Lab-on-a- Chip) has emerged as a technology that might be able to satisfy this need. In generic terms, a biochip is a miniaturized microfluidic device analogous to the electronic microchip that ushered in the digital age. It consists of tiny microfluidic channels, pumps and valves that transport small amounts of sample fluids to biosensors that can perform a variety of tests on those fluids in near real time. It has the obvious advantages of being small, lightweight, requiring less sample fluids and reagents and being more sensitive and efficient than larger devices currently in use. Some of the desired space-based applications would be to provide smaller, more robust devices for analyzing blood, saliva and urine and for testing water and food supplies for the presence of harmful contaminants and microorganisms. Our group has undertaken the goal of adapting as well as improving upon current biochip technology for use in long-duration microgravity environments. In addition to developing computational models of the microfluidic channels, valves and pumps that form the basis of every biochip, we are also trying to identify potential problems that could arise in reduced gravity and develop solutions to these problems. One such problem is due to the prevalence of bubbly sample fluids in microgravity. A bubble trapped in a microfluidic channel could be detrimental to the operation of a biochip. Therefore, the process of bubble formation in microgravity needs to be studied, and a model of this process has been developed and used to understand how bubbles develop and move through biochip components. It is clear that some type of bubble filter would be necessary in Space, and

  12. Machine vision for digital microfluidics.

    PubMed

    Shin, Yong-Jun; Lee, Jeong-Bong

    2010-01-01

    Machine vision is widely used in an industrial environment today. It can perform various tasks, such as inspecting and controlling production processes, that may require humanlike intelligence. The importance of imaging technology for biological research or medical diagnosis is greater than ever. For example, fluorescent reporter imaging enables scientists to study the dynamics of gene networks with high spatial and temporal resolution. Such high-throughput imaging is increasingly demanding the use of machine vision for real-time analysis and control. Digital microfluidics is a relatively new technology with expectations of becoming a true lab-on-a-chip platform. Utilizing digital microfluidics, only small amounts of biological samples are required and the experimental procedures can be automatically controlled. There is a strong need for the development of a digital microfluidics system integrated with machine vision for innovative biological research today. In this paper, we show how machine vision can be applied to digital microfluidics by demonstrating two applications: machine vision-based measurement of the kinetics of biomolecular interactions and machine vision-based droplet motion control. It is expected that digital microfluidics-based machine vision system will add intelligence and automation to high-throughput biological imaging in the future.

  13. Microfluidic chip integrated with flexible PDMS-based electrochemical cytosensor for dynamic analysis of drug-induced apoptosis on HeLa cells.

    PubMed

    Cao, Jun-Tao; Zhu, Ying-Di; Rana, Rohit Kumar; Zhu, Jun-Jie

    2014-01-15

    A novel microfluidic platform integrated with a flexible PDMS-based electrochemical cytosensor was developed for real-time monitoring of the proliferation and apoptosis of HeLa cells. The PDMS-gold film, which had a conductive smooth surface and was semi-transparent, facilitated electrochemical measurements and optical microscope observations. We observed distinct increases and decreases in peak current intensity, corresponding to cell proliferation in culture medium and apoptosis in the presence of an anticancer drug, respectively. This electrochemical analysis method permitted real-time, label-free monitoring of cell behavior, and the electrochemical results were confirmed with optical microscopy. The flexible microfluidic electrochemical platform presented here is suitable for on-site monitoring of cell behavior in microenvironments.

  14. 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).

  15. End-of-life nursing education consortium for pediatric palliative care (ELNEC-PPC).

    PubMed

    Malloy, Pam; Sumner, Elizabeth; Virani, Rose; Ferrell, Betty

    2007-01-01

    Pediatric nurses must often care for children with life-threatening illness. Although the child may be a neonate with multiple organ failure, a young adolescent diagnosed with HIV, or a 7-year-old child involved in a serious bicycle accident, pediatric nurses are an essential part of the interdisciplinary team that plans, organizes, implements, and manages the care of these children and their families. To date, more than 600 pediatric nurses have attended a national End-of-Life Nursing Education Consortium-Pediatric Palliative Care (ELNEC-PPC) training program. Many of these nurses have returned to their institutions dedicated to making a difference in the palliative care provided to children and their families. Because pediatric palliative care education is so important, many trainers have incorporated ELNEC-PPC into their nursing orientation, annual competencies, and undergraduate and graduate nursing education. They are developing standards of care and serve on key hospital/hospice committees, such as policy, education, clinical care, and ethics committees. This article showcases various activities of ELNEC-PPC trainers and demonstrates their commitment to improve pediatric palliative care not only in their institutions but also on local, state, national, and international levels.

  16. Injection molded microfluidic devices for biological sample separation and detection

    NASA Astrophysics Data System (ADS)

    Morales, Alfredo M.; Simmons, Blake A.; Wallow, Thomas I.; Campbell, K. Jeffery; Mani, Seethambal S.; Mittal, Brita; Crocker, Robert W.; Cummings, Eric B.; Davalos, Rafael V.; Domeier, Linda A.; Hunter, Marion C.; Krafcik, Karen L.; McGraw, Gregory J.; Mosier, Bruce P.; Sickafoose, Shane M.

    2006-01-01

    We are developing a variety of microsystems for the separation and detection of biological samples. At the heart of these systems, inexpensive polymer microfluidic chips carry out sample preparation and analysis. Fabrication of polymer microfluidic chips involves the creation of a master in etched silicon or glass; plating of the master to produce a nickel stamp; large lot chip replication by injection molding; precision chip sealing; and chemical modification of channel surfaces. Separation chips rely on insulator-based dielectrophoresis for the separation of biological particles. Detection chips carry out capillary electrophoresis to detect fluorescent tags that identify specific biological samples. Since the performance and reliability of these microfluidic chips are very sensitive to fluidic impedance, electromagnetic flux, and zeta potential, the microchannel dimensions, shape, and surface chemistry have to be tightly controlled during chip fabrication and use. This paper will present an overview of chip design, fabrication, and testing. Dimensional metrology data, surface chemistry characterization, and chip performance data will be discussed in detail.

  17. 3D printed microfluidics for biological applications.

    PubMed

    Ho, Chee Meng Benjamin; Ng, Sum Huan; Li, King Ho Holden; Yoon, Yong-Jin

    2015-01-01

    The term "Lab-on-a-Chip," is synonymous with describing microfluidic devices with biomedical applications. Even though microfluidics have been developing rapidly over the past decade, the uptake rate in biological research has been slow. This could be due to the tedious process of fabricating a chip and the absence of a "killer application" that would outperform existing traditional methods. In recent years, three dimensional (3D) printing has been drawing much interest from the research community. It has the ability to make complex structures with high resolution. Moreover, the fast building time and ease of learning has simplified the fabrication process of microfluidic devices to a single step. This could possibly aid the field of microfluidics in finding its "killer application" that will lead to its acceptance by researchers, especially in the biomedical field. In this paper, a review is carried out of how 3D printing helps to improve the fabrication of microfluidic devices, the 3D printing technologies currently used for fabrication and the future of 3D printing in the field of microfluidics.

  18. Microfluidic CODES: a scalable multiplexed electronic sensor for orthogonal detection of particles in microfluidic channels.

    PubMed

    Liu, Ruxiu; Wang, Ningquan; Kamili, Farhan; Sarioglu, A Fatih

    2016-04-21

    Numerous biophysical and biochemical assays rely on spatial manipulation of particles/cells as they are processed on lab-on-a-chip devices. Analysis of spatially distributed particles on these devices typically requires microscopy negating the cost and size advantages of microfluidic assays. In this paper, we introduce a scalable electronic sensor technology, called microfluidic CODES, that utilizes resistive pulse sensing to orthogonally detect particles in multiple microfluidic channels from a single electrical output. Combining the techniques from telecommunications and microfluidics, we route three coplanar electrodes on a glass substrate to create multiple Coulter counters producing distinct orthogonal digital codes when they detect particles. We specifically design a digital code set using the mathematical principles of Code Division Multiple Access (CDMA) telecommunication networks and can decode signals from different microfluidic channels with >90% accuracy through computation even if these signals overlap. As a proof of principle, we use this technology to detect human ovarian cancer cells in four different microfluidic channels fabricated using soft lithography. Microfluidic CODES offers a simple, all-electronic interface that is well suited to create integrated, low-cost lab-on-a-chip devices for cell- or particle-based assays in resource-limited settings.

  19. MEMS and microfluidics for diagnostics devices.

    PubMed

    Rosen, Y; Gurman, P

    2010-06-01

    There are conditions in clinical medicine demanding critical therapeutic decisions. These conditions necessitate accuracy, rapidity, accessibility, cost-effectiveness and mobility. New technologies have been developed in order to address these challenges. Microfluidics and Micro Electro-Mechanical Systems are two of such technologies. Microfluidics, a discipline that involves processing fluids at the microscale in etched microchannels, is being used to build lab- on-a-chip systems to run chemical and biological assays. These systems are being transformed into handheld devices designed to be used at remote