Inexpensive and Safe DNA Gel Electrophoresis Using Household Materials

ERIC Educational Resources Information Center

Ens, S.; Olson, A. B.; Dudley, C.; Ross, N. D., III; Siddiqi, A. A.; Umoh, K. M.; Schneegurt, M. A.

2012-01-01

Gel electrophoresis is the single most important molecular biology technique and it is central to life sciences research, but it is often too expensive for the secondary science classroom or homeschoolers. A simple safe low-cost procedure is described here that uses household materials to construct and run DNA gel electrophoresis. Plastic…

Improvement of J(sc) in a Cu2ZnSnS4 Solar Cell by Using a Thin Carbon Intermediate Layer at the Cu2ZnSnS4/Mo Interface.

PubMed

Zhou, Fangzhou; Zeng, Fangqin; Liu, Xu; Liu, Fangyang; Song, Ning; Yan, Chang; Pu, Aobo; Park, Jongsung; Sun, Kaiwen; Hao, Xiaojing

2015-10-21

Back contact modification plays an important role in improving energy conversion efficiency of Cu2ZnSnS4 (CZTS) thin film solar cells. In this paper, an ultrathin carbon layer is introduced on molybdenum (Mo)-coated soda lime glass (SLG) prior to the deposition of CZTS precursor to improve the back contact and therefore enhance CZTS solar cell efficiency. By introducing this layer, the short circuit current (Jsc) and device conversion efficiency increase for both nonvacuum (sol-gel) and vacuum (sputtering) methods. Specifically, for the sol-gel based process, Jsc increases from 13.60 to 16.96 mA/cm(2) and efficiency from 4.47% to 5.52%, while for the sputtering based process, Jsc increases from 17.50 to 20.50 mA/cm(2) and efficiency from 4.10% to 5.20%. Furthermore, introduction of this layer does not lead to any deterioration of either open circuit voltage (Voc) or fill factor (FF).

Procedures and computer program for deriving the Ferguson plot from electrophoresis in a single pore gradient gel: application to agarose gel and a polystyrene particle.

PubMed

Tietz, D; Gombocz, E; Chrambach, A

1991-10-01

This study presents a computerized evaluation of pore gradient gel electrophoretograms to arrive at estimates for both the particle-free mobility and retardation coefficient, which is related to particle size. Agarose pore gradient gels ranging from 0.2 to 1.1% agarose were formed. Gel gradients were stabilized during their formation by a density gradient of 0-20% 5-(N-2,3-dihydroxypropylacetamido)- 2,4,6-triiodo-N,N'bis-(2,3-dihydroxypropyl)-isophthalamide (Nycodenz). Densitometry of gelled-in Bromophenol Blue showed that these pore gradients exhibited a linear central segment and were reproducible. Migration distances of polystyrene sulfate microspheres (36.5 nm radius) in agarose pore gradient gel electrophoresis were determined by time-lapse photography at several durations of electrophoresis. These migration distances were evaluated as a function of migration time as previously reported (D. Tietz, Adv. Electrophoresis 1988, 2, 109-169). Although this is not necessarily required, the mathematical approach used in this study assumed linearity of both the pore gradient and the Ferguson plot for reasons of simplicity. The data evaluation on the basis of the extended Ogston model is incorporated in a user-friendly program, GRADFIT, which is designed for personal computers (Macintosh). The results obtained are compared with (1) conventional electrophoresis using several gels of single concentration with and without Nycodenz, and (ii) a different mathematical approach for the analysis of gradient gels (Rodbard et al., Anal. Biochem. 1971, 40, 135-157). Moreover, a simple procedure for evaluating linear pore gradient gels using linear regression analysis is presented. It is concluded that the values of particle-free mobility and retardation coefficient derived from pore gradient gel electrophoresis using the different mathematical methods are statistically indistinguishable from each other. However, these values are different, albeit close, to those obtained from conventional Ferguson plots. One of the possible reasons for this relatively minor discrepancy is that the particle-free mobility changed slightly during electrophoresis, which has a different effect on electrophoresis in homogeneous gels (single time measurement) and pore gradient gels (multiple time measurements). The characterization of particles according to size and charge by pore gradient electrophoresis provides a significant operational simplification and sample economy compared to that requiring the use of several gel concentrations, although at the price of increased requirements of instrumentation.

Ink-native electrophoresis: an alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity.

PubMed

Kaneko, Keisuke; Sueyoshi, Noriyuki; Kameshita, Isamu; Ishida, Atsuhiko

2013-09-15

Blue-native electrophoresis (BNE) is a useful technique for analyzing protein complexes, but the Coomassie brilliant blue (CBB) dye used in BNE often hampers in-gel detection of enzymatic activity. Here we report an improved method, termed ink-native electrophoresis (INE), in which Pelikan 4001 fountain pen ink is used as a charge-shifting agent instead of CBB. INE is more suitable than BNE for in-gel detection of protein kinase activity after polyacrylamide gel electrophoresis (PAGE), and its performance in protein complex separation is comparable to that of conventional BNE. INE may provide a powerful tool to isolate and analyze various protein complexes. Copyright © 2013 Elsevier Inc. All rights reserved.

Atomic layer deposition of ultrathin blocking layer for low-temperature solid oxide fuel cell on nanoporous substrate

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

Yu, Wonjong; Cho, Gu Young; Noh, Seungtak

2015-01-15

An ultrathin yttria-stabilized zirconia (YSZ) blocking layer deposited by atomic layer deposition (ALD) was utilized for improving the performance and reliability of low-temperature solid oxide fuel cells (SOFCs) supported by an anodic aluminum oxide substrate. Physical vapor-deposited YSZ and gadolinia-doped ceria (GDC) electrolyte layers were deposited by a sputtering method. The ultrathin ALD YSZ blocking layer was inserted between the YSZ and GDC sputtered layers. To investigate the effects of an inserted ultrathin ALD blocking layer, SOFCs with and without an ultrathin ALD blocking layer were electrochemically characterized. The open circuit voltage (1.14 V) of the ALD blocking-layered SOFC was visiblymore » higher than that (1.05 V) of the other cell. Furthermore, the ALD blocking layer augmented the power density and improved the reproducibility.« less

Copolymers For Capillary Gel Electrophoresis

DOEpatents

Liu, Changsheng; Li, Qingbo

2005-08-09

This invention relates to an electrophoresis separation medium having a gel matrix of at least one random, linear copolymer comprising a primary comonomer and at least one secondary comonomer, wherein the comonomers are randomly distributed along the copolymer chain. The primary comonomer is an acrylamide or an acrylamide derivative that provides the primary physical, chemical, and sieving properties of the gel matrix. The at least one secondary comonomer imparts an inherent physical, chemical, or sieving property to the copolymer chain. The primary and secondary comonomers are present in a ratio sufficient to induce desired properties that optimize electrophoresis performance. The invention also relates to a method of separating a mixture of biological molecules using this gel matrix, a method of preparing the novel electrophoresis separation medium, and a capillary tube filled with the electrophoresis separation medium.

Chromatographic and electrophoretic approaches in ink analysis.

PubMed

Zlotnick, J A; Smith, F P

1999-10-15

Inks are manufactured from a wide variety of substances that exhibit very different chemical behaviors. Inks designed for use in different writing instruments or printing methods have quite dissimilar components. Since the 1950s chromatographic and electrophoretic methods have played important roles in the analysis of inks, where compositional information may have bearing on the investigation of counterfeiting, fraud, forgery, and other crimes. Techniques such as paper chromatography and electrophoresis, thin-layer chromatography, high-performance liquid chromatography, gas chromatography, gel electrophoresis, and the relatively new technique of capillary electrophoresis have all been explored as possible avenues for the separation of components of inks. This paper reviews the components of different types of inks and applications of the above separation methods are reviewed.

Using Gel Electrophoresis To Illustrate Protein Diversity and Isoelectric Point.

ERIC Educational Resources Information Center

Browning, Mark; Vanable, Joseph

2002-01-01

Demonstrates the differences in protein structures by focusing on isoelectric point with an experiment that is observable under certain pH levels in gel electrophoresis. Explains the electrophoresis procedure and reports results of the experiments. (YDS)

Getting the Most out of Electrophoresis Units

ERIC Educational Resources Information Center

Mulvihill, Charlotte

2007-01-01

At Oklahoma City Community College, they have developed gel electrophoresis activities that support active learning of many scientific concepts, including: pH, electrolysis, oxidation reduction, electrical currents, potentials, conductivity, molarity, gel electrophoresis, DNA and protein separation, and DNA fingerprinting. This article presents…

Analyzing genetic diversity in conifers...isozyme resolution by starch gel electrophoresis

Treesearch

M. Thompson Conkle

1972-01-01

Enzymes in forest tree materials can be resolved by starch gel electrophoresis. A gel slab is prepared in a mold assembled from glass and plastic. Wicks containing an aqueous extract of macerated plant material are inserted in the gel and processed. The gel is sliced, stained, examined, and photographed. Isozyme bands produced by differential migration of enzymes...

Visualization of yeast chromosomal DNA

NASA Technical Reports Server (NTRS)

Lubega, Seth

1990-01-01

The DNA molecule is the most significant life molecule since it codes the blue print for other structural and functional molecules of all living organisms. Agarose gel electrophoresis is now being widely used to separate DNA of virus, bacteria, and lower eukaryotes. The task was undertaken of reviewing the existing methods of DNA fractionation and microscopic visualization of individual chromosonal DNA molecules by gel electrophoresis as a basis for a proposed study to investigate the feasibility of separating DNA molecules in free fluids as an alternative to gel electrophoresis. Various techniques were studied. On the molecular level, agarose gel electrophoresis is being widely used to separate chromosomal DNA according to molecular weight. Carl and Olson separate and characterized the entire karyotype of a lab strain of Saccharomyces cerevisiae. Smith et al. and Schwartz and Koval independently reported the visualization of individual DNA molecules migrating through agarose gel matrix during electrophoresis. The techniques used by these researchers are being reviewed in the lab as a basis for the proposed studies.

In-gel staining of proteins in native polyacrylamide gel electrophoresis using meso-tetrakis(4-sulfonatophenyl) porphyrin.

PubMed

Divakar, K; Devi, G Nandhini; Gautam, Pennathur

2012-01-01

Protein identification in polyacrylamide gel electrophoresis (PAGE) requires post-electrophoretic steps like fixing, staining, and destaining of the gel, which are time-consuming and cumbersome. A new method for direct visualization of protein bands in PAGE has been developed using meso-tetrakis(4-sulfonatophenyl)porphyrin (TPPS) as a dye without the need for any post-electrophoretic steps; thus, separation and recovery of enzymes become much easier for further analysis. Activity staining was carried out to show that the biochemical activity of the enzymes was preserved after electrophoresis.

Agarose gel electrophoresis for the separation of DNA fragments.

PubMed

Lee, Pei Yun; Costumbrado, John; Hsu, Chih-Yuan; Kim, Yong Hoon

2012-04-20

Agarose gel electrophoresis is the most effective way of separating DNA fragments of varying sizes ranging from 100 bp to 25 kb(1). Agarose is isolated from the seaweed genera Gelidium and Gracilaria, and consists of repeated agarobiose (L- and D-galactose) subunits(2). During gelation, agarose polymers associate non-covalently and form a network of bundles whose pore sizes determine a gel's molecular sieving properties. The use of agarose gel electrophoresis revolutionized the separation of DNA. Prior to the adoption of agarose gels, DNA was primarily separated using sucrose density gradient centrifugation, which only provided an approximation of size. To separate DNA using agarose gel electrophoresis, the DNA is loaded into pre-cast wells in the gel and a current applied. The phosphate backbone of the DNA (and RNA) molecule is negatively charged, therefore when placed in an electric field, DNA fragments will migrate to the positively charged anode. Because DNA has a uniform mass/charge ratio, DNA molecules are separated by size within an agarose gel in a pattern such that the distance traveled is inversely proportional to the log of its molecular weight(3). The leading model for DNA movement through an agarose gel is "biased reptation", whereby the leading edge moves forward and pulls the rest of the molecule along(4). The rate of migration of a DNA molecule through a gel is determined by the following: 1) size of DNA molecule; 2) agarose concentration; 3) DNA conformation(5); 4) voltage applied, 5) presence of ethidium bromide, 6) type of agarose and 7) electrophoresis buffer. After separation, the DNA molecules can be visualized under uv light after staining with an appropriate dye. By following this protocol, students should be able to: Understand the mechanism by which DNA fragments are separated within a gel matrix Understand how conformation of the DNA molecule will determine its mobility through a gel matrix Identify an agarose solution of appropriate concentration for their needs Prepare an agarose gel for electrophoresis of DNA samples Set up the gel electrophoresis apparatus and power supply Select an appropriate voltage for the separation of DNA fragments Understand the mechanism by which ethidium bromide allows for the visualization of DNA bands Determine the sizes of separated DNA fragments.

A sensitive gel-based global O-glycomics approach reveals high levels of mannosyl glycans in the high mass region of the mouse brain proteome.

PubMed

Breloy, Isabelle; Pacharra, Sandra; Aust, Christina; Hanisch, Franz-Georg

2012-08-01

We developed a gel-based global O-glycomics method applicable for highly complex protein mixtures entrapped in discontinuous gradient gel layers. The protocol is based on in-gel proteolysis with pronase followed by (glyco)peptide elution and off-gel reductive β-elimination. The protocol offers robust performance with sensitivity in the low picomolar range, is compatible with gel-based proteomics, and shows superior performance in global applications in comparison with workflows eliminating glycans in-gel or from electroblotted glycoproteins. By applying this method, we analyzed the O-glycome of human myoblasts and of the mouse brain O-glycoproteome. After semipreparative separation of mouse brain proteins by one-dimensional SDS gel electrophoresis, the O-glycans from proteins in different mass ranges were characterized with a focus on O-mannose-based glycans. The relative proportion of the latter, which generally represent a rare modification, increases to comparatively high levels in the mouse brain proteome in dependence of increasing protein masses.

Automatic DNA Diagnosis for 1D Gel Electrophoresis Images using Bio-image Processing Technique.

PubMed

Intarapanich, Apichart; Kaewkamnerd, Saowaluck; Shaw, Philip J; Ukosakit, Kittipat; Tragoonrung, Somvong; Tongsima, Sissades

2015-01-01

DNA gel electrophoresis is a molecular biology technique for separating different sizes of DNA fragments. Applications of DNA gel electrophoresis include DNA fingerprinting (genetic diagnosis), size estimation of DNA, and DNA separation for Southern blotting. Accurate interpretation of DNA banding patterns from electrophoretic images can be laborious and error prone when a large number of bands are interrogated manually. Although many bio-imaging techniques have been proposed, none of them can fully automate the typing of DNA owing to the complexities of migration patterns typically obtained. We developed an image-processing tool that automatically calls genotypes from DNA gel electrophoresis images. The image processing workflow comprises three main steps: 1) lane segmentation, 2) extraction of DNA bands and 3) band genotyping classification. The tool was originally intended to facilitate large-scale genotyping analysis of sugarcane cultivars. We tested the proposed tool on 10 gel images (433 cultivars) obtained from polyacrylamide gel electrophoresis (PAGE) of PCR amplicons for detecting intron length polymorphisms (ILP) on one locus of the sugarcanes. These gel images demonstrated many challenges in automated lane/band segmentation in image processing including lane distortion, band deformity, high degree of noise in the background, and bands that are very close together (doublets). Using the proposed bio-imaging workflow, lanes and DNA bands contained within are properly segmented, even for adjacent bands with aberrant migration that cannot be separated by conventional techniques. The software, called GELect, automatically performs genotype calling on each lane by comparing with an all-banding reference, which was created by clustering the existing bands into the non-redundant set of reference bands. The automated genotype calling results were verified by independent manual typing by molecular biologists. This work presents an automated genotyping tool from DNA gel electrophoresis images, called GELect, which was written in Java and made available through the imageJ framework. With a novel automated image processing workflow, the tool can accurately segment lanes from a gel matrix, intelligently extract distorted and even doublet bands that are difficult to identify by existing image processing tools. Consequently, genotyping from DNA gel electrophoresis can be performed automatically allowing users to efficiently conduct large scale DNA fingerprinting via DNA gel electrophoresis. The software is freely available from http://www.biotec.or.th/gi/tools/gelect.

Simple and rapid system for two-dimensional gel electrophoresis technique: A laboratory exercise for high school students.

PubMed

Maurye, Praveen; Basu, Arpita; Biswas, Jayanta Kumar; Bandyopadhyay, Tapas Kumar; Naskar, Malay

2018-02-28

Polyacrylamide gel electrophoresis (PAGE) is the most classical technique favored worldwide for resolution of macromolecules in many biochemistry laboratories due to its incessant advanced developments and wide modifications. These ever-growing advancements in the basic laboratory equipments lead to emergence of many expensive, complex, and tricky laboratory equipments. Practical courses of biochemistry at high school or undergraduate levels are often affected by these complications. Two dimensional gel electrophoresis technique (2D-PAGE) used for resolving thousands of proteins in a gel is a combination of isoelectric focusing (first dimension gel electrophoresis technique) and sodium-dodecylsulphate PAGE (second dimension gel electrophoresis technique or SDS-PAGE). Two different laboratory equipments are needed to carry out effective 2D-PAGE technique, which also invites extra burden to the school laboratory. Here, we describe a low cost, time saving and simple gel cassette for protein 2D-PAGE technique that uses easily fabricated components and routine off-the-shelf materials. The performance of the apparatus was verified in a practical exercise by a group of high school students with positive outcomes. © 2018 by The International Union of Biochemistry and Molecular Biology, 2018. © 2018 The International Union of Biochemistry and Molecular Biology.

Purification and characterization of DR_2577 (SlpA) a major S-layer protein from Deinococcus radiodurans

PubMed Central

Farci, Domenica; Bowler, Matthew W.; Esposito, Francesca; McSweeney, Sean; Tramontano, Enzo; Piano, Dario

2015-01-01

The protein DR_2577 is a major Surface layer component of the radio-resistant bacterium Deinococcus radiodurans. In the present study DR_2577 has been purified and its oligomeric profile characterized by means of size exclusion chromatography and gel electrophoresis. DR_2577 was found to be organized into three hierarchical orders characterized by monomers, stable dimers formed by the occurrence of disulfide bonds, and hexamers resulting from a combination of dimers. The structural implications of these findings are discussed providing new elements for a more integrated model of this S-layer. PMID:26074883

Purification and characterization of DR_2577 (SlpA) a major S-layer protein from Deinococcus radiodurans

DOE PAGES

Farci, Domenica; Bowler, Matthew W.; Esposito, Francesca; ...

2015-06-03

The protein DR_2577 is a major Surface layer component of the radio-resistant bacterium Deinococcus radiodurans. In the present study DR_2577 has been purified and its oligomeric profile characterized by means of size exclusion chromatography and gel electrophoresis. DR_2577 was found to be organized into three hierarchical orders characterized by monomers, stable dimers formed by the occurrence of disulfide bonds, and hexamers resulting from a combination of dimers. Finally, the structural implications of these findings are discussed providing new elements for a more integrated model of this S-layer.

Performing Isoelectric Focusing and Simultaneous Fractionation of Proteins on A Rotary Valve Followed by Sodium Dodecyl – Polyacrylamide Gel Electrophoresis

PubMed Central

Wang, Wei; Lu, Joann J.; Gu, Congying; Zhou, Lei; Liu, Shaorong

2013-01-01

In this technical note, we design and fabricate a novel rotary valve and demonstrate its feasibility for performing isoelectric focusing and simultaneous fractionation of proteins, followed by sodium dodecyl – polyacrylamide gel electrophoresis. The valve has two positions. In one position, the valve routes a series of capillary loops together into a single capillary tube where capillary isoelectric focusing (CIEF) is performed. By switching the valve to another position, the CIEF-resolved proteins in all capillary loops are isolated simultaneously, and samples in the loops are removed and collected in vials. After the collected samples are briefly processed, they are separated via sodium dodecyl – polyacrylamide gel electrophoresis (SDS-PAGE, the 2nd-D separation) on either a capillary gel electrophoresis instrument or a slab-gel system. The detailed valve configuration is illustrated, and the experimental conditions and operation protocols are discussed. PMID:23819755

A Simple Vertical Slab Gel Electrophoresis Apparatus.

ERIC Educational Resources Information Center

Carter, J. B.; And Others

1983-01-01

Describes an inexpensive, easily constructed, and safe vertical slab gel kit used routinely for sodium dodecyl sulphate-polyacrylamide gel electrophoresis research and student experiments. Five kits are run from a single transformer. Because toxic solutions are used, students are given plastic gloves and closely supervised during laboratory…

Supramolecular gel electrophoresis of large DNA fragments.

PubMed

Tazawa, Shohei; Kobayashi, Kazuhiro; Oyoshi, Takanori; Yamanaka, Masamichi

2017-10-01

Pulsed-field gel electrophoresis is a frequent technique used to separate exceptionally large DNA fragments. In a typical continuous field electrophoresis, it is challenging to separate DNA fragments larger than 20 kbp because they migrate at a comparable rate. To overcome this challenge, it is necessary to develop a novel matrix for the electrophoresis. Here, we describe the electrophoresis of large DNA fragments up to 166 kbp using a supramolecular gel matrix and a typical continuous field electrophoresis system. C 3 -symmetric tris-urea self-assembled into a supramolecular hydrogel in tris-boric acid-EDTA buffer, a typical buffer for DNA electrophoresis, and the supramolecular hydrogel was used as a matrix for electrophoresis to separate large DNA fragments. Three types of DNA marker, the λ-Hind III digest (2 to 23 kbp), Lambda DNA-Mono Cut Mix (10 to 49 kbp), and Marker 7 GT (10 to 165 kbp), were analyzed in this study. Large DNA fragments of greater than 100 kbp showed distinct mobility using a typical continuous field electrophoresis system. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Gel Electrophoresis--The Easy Way for Students

ERIC Educational Resources Information Center

VanRooy, Wilhelmina; Sultana, Khalida

2010-01-01

This article describes a simple, inexpensive, easy to conduct gel-electrophoresis activity using food dyes. It is an alternative to the more expensive counterparts which require agarose gel, DNA samples, purchased chamber and Tris-borate-EDTA buffer. We suggest some learning activities for senior biology students along with comments on several…

Gel Electrophoresis on a Budget to Dye for

ERIC Educational Resources Information Center

Yu, Julie H.

2010-01-01

Gel electrophoresis is one of the most important tools used in molecular biology and has facilitated the entire field of genetic engineering by enabling the separation of nucleic acids and proteins. However, commercial electrophoresis kits can cost up to $800 for each setup, which is cost prohibitive for most classroom budgets. This article…

Curie temperature of ultrathin ferromagnetic layer with Dzyaloshinskii-Moriya interaction

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

You, Chun-Yeol

2014-08-07

We investigate the effect of the Dzyaloshinskii-Moriya interaction (DMI) on the Curie temperature of the ultrathin ferromagnetic layers. It has been known that the Curie temperature of the ferromagnet depends on spin wave excitation energies, and they are affected by DMI. Therefore, the ferromagnetic transition temperature of the ultrathin ferromagnetic layer must be sensitive on the DMI. We find that the Curie temperature depends on the DMI by using the double time Green's function method. Since the DMI is arisen by the inversion symmetry breaking structure, the DMI is always important in the inversion symmetry breaking ultrathin ferromagnetic layers.

Integrated protocol for reliable and fast quantification and documentation of electrophoresis gels.

PubMed

Rehbein, Peter; Schwalbe, Harald

2015-06-01

Quantitative analysis of electrophoresis gels is an important part in molecular cloning, as well as in protein expression and purification. Parallel quantifications in yield and purity can be most conveniently obtained from densitometric analysis. This communication reports a comprehensive, reliable and simple protocol for gel quantification and documentation, applicable for single samples and with special features for protein expression screens. As major component of the protocol, the fully annotated code of a proprietary open source computer program for semi-automatic densitometric quantification of digitized electrophoresis gels is disclosed. The program ("GelQuant") is implemented for the C-based macro-language of the widespread integrated development environment of IGOR Pro. Copyright © 2014 Elsevier Inc. All rights reserved.

DNA DAMAGE QUANTITATION BY ALKALINE GEL ELECTROPHORESIS.

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

SUTHERLAND,B.M.; BENNETT,P.V.; SUTHERLAND, J.C.

2004-03-24

Physical and chemical agents in the environment, those used in clinical applications, or encountered during recreational exposures to sunlight, induce damages in DNA. Understanding the biological impact of these agents requires quantitation of the levels of such damages in laboratory test systems as well as in field or clinical samples. Alkaline gel electrophoresis provides a sensitive (down to {approx} a few lesions/5Mb), rapid method of direct quantitation of a wide variety of DNA damages in nanogram quantities of non-radioactive DNAs from laboratory, field, or clinical specimens, including higher plants and animals. This method stems from velocity sedimentation studies of DNAmore » populations, and from the simple methods of agarose gel electrophoresis. Our laboratories have developed quantitative agarose gel methods, analytical descriptions of DNA migration during electrophoresis on agarose gels (1-6), and electronic imaging for accurate determinations of DNA mass (7-9). Although all these components improve sensitivity and throughput of large numbers of samples (7,8,10), a simple version using only standard molecular biology equipment allows routine analysis of DNA damages at moderate frequencies. We present here a description of the methods, as well as a brief description of the underlying principles, required for a simplified approach to quantitation of DNA damages by alkaline gel electrophoresis.« less

Starch gel electrophoresis of conifer seeds: a laboratory manual

Treesearch

M. Thompson Conkle; Paul D. Hodgskiss; Lucy B. Nunnally; Serena C. Hunter

1982-01-01

This manual describes fast, low-cost biochemical procedures for separating enzymes representing numerous genes of forest trees. During electrophoresis the mixture of enzymes from a megagametophyte or embryo of a germinated seed separates in a gel. Specific stains applied to gel slices locate each enzyme. These procedures expand on those developed for crops research....

Ultrathin nondoped emissive layers for efficient and simple monochrome and white organic light-emitting diodes.

PubMed

Zhao, Yongbiao; Chen, Jiangshan; Ma, Dongge

2013-02-01

In this paper, highly efficient and simple monochrome blue, green, orange, and red organic light emitting diodes (OLEDs) based on ultrathin nondoped emissive layers (EMLs) have been reported. The ultrathin nondoped EML was constructed by introducing a 0.1 nm thin layer of pure phosphorescent dyes between a hole transporting layer and an electron transporting layer. The maximum external quantum efficiencies (EQEs) reached 17.1%, 20.9%, 17.3%, and 19.2% for blue, green, orange, and red monochrome OLEDs, respectively, indicating the universality of the ultrathin nondoped EML for most phosphorescent dyes. On the basis of this, simple white OLED structures are also demonstrated. The demonstrated complementary blue/orange, three primary blue/green/red, and four color blue/green/orange/red white OLEDs show high efficiency and good white emission, indicating the advantage of ultrathin nondoped EMLs on constructing simple and efficient white OLEDs.

All-solid-state flexible ultrathin micro-supercapacitors based on graphene.

PubMed

Niu, Zhiqiang; Zhang, Li; Liu, Lili; Zhu, Bowen; Dong, Haibo; Chen, Xiaodong

2013-08-07

Flexible, compact, ultrathin and all-solid-state micro-supercapacitors are prepared by coating H₃PO₄/PVA gel electrolyte onto micro-patterned rGO interdigitated electrodes prepared by combining photolithography with selective electrophoretic deposition. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Running DNA Mini-Gels in 20 Minutes or Less Using Sodium Boric Acid Buffer

ERIC Educational Resources Information Center

Jenkins, Kristin P.; Bielec, Barbara

2006-01-01

Providing a biotechnology experience for students can be challenging on several levels, and time is a real constraint for many experiments. Many DNA based methods require a gel electrophoresis step, and although some biotechnology procedures have convenient break points, gel electrophoresis does not. In addition to the time required for loading…

Synthesis of hydrogel via click chemistry for DNA electrophoresis.

PubMed

Finetti, Chiara; Sola, Laura; Elliott, Jim; Chiari, Marcella

2017-09-01

This work introduces a novel sieving gel for DNA electrophoresis using a classical click chemistry reaction, the copper (I)-catalyzed azide-alkyne cycloaddition (CuAAC), to cross-link functional polymer chains. The efficiency of this reaction provides, under mild conditions, hydrogels with near-ideal network connectivity and improved physical properties. Hydrogel formation via click chemistry condensation of functional polymers does not involve the use of toxic monomers and UV initiation. The performance of the new hydrogel in the separation of double stranded DNA fragments was evaluated in the 2200 TapeStation system, an analytical platform, recently introduced by Agilent that combines the advantages of CE in terms of miniaturization and automation with the simplicity of use of slab gel electrophoresis. The click gel enables addition of florescent dyes prior to electrophoresis with considerable improvement of resolution and separation efficiency over conventional cross-linked polyacrylamide gels. Copyright © 2017 Elsevier B.V. All rights reserved.

Gel Electrophoresis of Gold-DNA Nano-Conjugates

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

Pellegrino, T.; Sperling, R.A.; Alivisatos, A.P.

2006-01-10

Single stranded DNA of different lengths and different amounts was attached to colloidal phosphine stabilized Au nanoparticles. The resulting conjugates were investigated in detail by a gel electrophoresis study based on 1200 gels. We demonstrate how these experiments help to understand the binding of DNA to Au particles. In particular we compare specific attachment of DNA via gold-thiol bonds with nonspecific adsorption of DNA. The maximum number of DNA molecules that can be bound per particle was determined. We also compare several methods to used gel electrophoresis for investigating the effective diameter of DNA-Au conjugates, such as using a calibrationmore » curve of particles with known diameters and Ferguson plots.« less

Combining high-throughput MALDI-TOF mass spectrometry and isoelectric focusing gel electrophoresis for virtual 2D gel-based proteomics.

PubMed

Lohnes, Karen; Quebbemann, Neil R; Liu, Kate; Kobzeff, Fred; Loo, Joseph A; Ogorzalek Loo, Rachel R

2016-07-15

The virtual two-dimensional gel electrophoresis/mass spectrometry (virtual 2D gel/MS) technology combines the premier, high-resolution capabilities of 2D gel electrophoresis with the sensitivity and high mass accuracy of mass spectrometry (MS). Intact proteins separated by isoelectric focusing (IEF) gel electrophoresis are imaged from immobilized pH gradient (IPG) polyacrylamide gels (the first dimension of classic 2D-PAGE) by matrix-assisted laser desorption/ionization (MALDI) MS. Obtaining accurate intact masses from sub-picomole-level proteins embedded in 2D-PAGE gels or in IPG strips is desirable to elucidate how the protein of one spot identified as protein 'A' on a 2D gel differs from the protein of another spot identified as the same protein, whenever tryptic peptide maps fail to resolve the issue. This task, however, has been extremely challenging. Virtual 2D gel/MS provides access to these intact masses. Modifications to our matrix deposition procedure improve the reliability with which IPG gels can be prepared; the new procedure is described. Development of this MALDI MS imaging (MSI) method for high-throughput MS with integrated 'top-down' MS to elucidate protein isoforms from complex biological samples is described and it is demonstrated that a 4-cm IPG gel segment can now be imaged in approximately 5min. Gel-wide chemical and enzymatic methods with further interrogation by MALDI MS/MS provide identifications, sequence-related information, and post-translational/transcriptional modification information. The MSI-based virtual 2D gel/MS platform may potentially link the benefits of 'top-down' and 'bottom-up' proteomics. Copyright © 2016 Elsevier Inc. All rights reserved.

A rapid high-resolution method for resolving DNA topoisomers.

PubMed

Mitchenall, Lesley A; Hipkin, Rachel E; Piperakis, Michael M; Burton, Nicolas P; Maxwell, Anthony

2018-01-16

Agarose gel electrophoresis has been the mainstay technique for the analysis of DNA samples of moderate size. In addition to separating linear DNA molecules, it can also resolve different topological forms of plasmid DNAs, an application useful for the analysis of the reactions of DNA topoisomerases. However, gel electrophoresis is an intrinsically low-throughput technique and suffers from other potential disadvantages. We describe the application of the QIAxcel Advanced System, a high-throughput capillary electrophoresis system, to separate DNA topoisomers, and compare this technique with gel electrophoresis. We prepared a range of topoisomers of plasmids pBR322 and pUC19, and a 339 bp DNA minicircle, and compared their separation by gel electrophoresis and the QIAxcel System. We found superior resolution with the QIAxcel System, and that quantitative analysis of topoisomer distributions was straightforward. We show that the QIAxcel system has advantages in terms of speed, resolution and cost, and can be applied to DNA circles of various sizes. It can readily be adapted for use in compound screening against topoisomerase targets.

Comparative Skeletal Muscle Proteomics Using Two-Dimensional Gel Electrophoresis

PubMed Central

Murphy, Sandra; Dowling, Paul; Ohlendieck, Kay

2016-01-01

The pioneering work by Patrick H. O’Farrell established two-dimensional gel electrophoresis as one of the most important high-resolution protein separation techniques of modern biochemistry (Journal of Biological Chemistry 1975, 250, 4007–4021). The application of two-dimensional gel electrophoresis has played a key role in the systematic identification and detailed characterization of the protein constituents of skeletal muscles. Protein changes during myogenesis, muscle maturation, fibre type specification, physiological muscle adaptations and natural muscle aging were studied in depth by the original O’Farrell method or slightly modified gel electrophoretic techniques. Over the last 40 years, the combined usage of isoelectric focusing in the first dimension and sodium dodecyl sulfate polyacrylamide slab gel electrophoresis in the second dimension has been successfully employed in several hundred published studies on gel-based skeletal muscle biochemistry. This review focuses on normal and physiologically challenged skeletal muscle tissues and outlines key findings from mass spectrometry-based muscle proteomics, which was instrumental in the identification of several thousand individual protein isoforms following gel electrophoretic separation. These muscle-associated protein species belong to the diverse group of regulatory and contractile proteins of the acto-myosin apparatus that forms the sarcomere, cytoskeletal proteins, metabolic enzymes and transporters, signaling proteins, ion-handling proteins, molecular chaperones and extracellular matrix proteins. PMID:28248237

Problem-Solving Test: Southwestern Blotting

ERIC Educational Resources Information Center

Szeberényi, József

2014-01-01

Terms to be familiar with before you start to solve the test: Southern blotting, Western blotting, restriction endonucleases, agarose gel electrophoresis, nitrocellulose filter, molecular hybridization, polyacrylamide gel electrophoresis, proto-oncogene, c-abl, Src-homology domains, tyrosine protein kinase, nuclear localization signal, cDNA,…

Disposable pen-shaped capillary gel electrophoresis cartridge for fluorescence detection of bio-molecules

NASA Astrophysics Data System (ADS)

Amirkhanian, Varoujan; Tsai, Shou-Kuan

2014-03-01

We introduce a novel and cost-effective capillary gel electrophoresis (CGE) system utilizing disposable pen-shaped gelcartridges for highly efficient, high speed, high throughput fluorescence detection of bio-molecules. The CGE system has been integrated with dual excitation and emission optical-fibers with micro-ball end design for fluorescence detection of bio-molecules separated and detected in a disposable pen-shaped capillary gel electrophoresis cartridge. The high-performance capillary gel electrophoresis (CGE) analyzer has been optimized for glycoprotein analysis type applications. Using commercially available labeling agent such as ANTS (8-aminonapthalene-1,3,6- trisulfonate) as an indicator, the capillary gel electrophoresis-based glycan analyzer provides high detection sensitivity and high resolving power in 2-5 minutes of separations. The system can hold total of 96 samples, which can be automatically analyzed within 4-5 hours. This affordable fiber optic based fluorescence detection system provides fast run times (4 minutes vs. 20 minutes with other CE systems), provides improved peak resolution, good linear dynamic range and reproducible migration times, that can be used in laboratories for high speed glycan (N-glycan) profiling applications. The CGE-based glycan analyzer will significantly increase the pace at which glycoprotein research is performed in the labs, saving hours of preparation time and assuring accurate, consistent and economical results.

Pulsed-field gel electrophoresis typing of Staphylococcus aureus isolates

USDA-ARS?s Scientific Manuscript database

Pulsed-field gel electrophoresis (PFGE) is the most applied and effective genetic typing method for epidemiological studies and investigation of foodborne outbreaks caused by different pathogens, including Staphylococcus aureus. The technique relies on analysis of large DNA fragments generated by th...

Ultrathin Coating of Confined Pt Nanocatalysts by Atomic Layer Deposition for Enhanced Catalytic Performance in Hydrogenation Reactions.

PubMed

Wang, Meihua; Gao, Zhe; Zhang, Bin; Yang, Huimin; Qiao, Yan; Chen, Shuai; Ge, Huibin; Zhang, Jiankang; Qin, Yong

2016-06-13

Metal-support interfaces play a prominent role in heterogeneous catalysis. However, tailoring the metal-support interfaces to realize full utilization remains a major challenge. In this work, we propose a graceful strategy to maximize the metal-oxide interfaces by coating confined nanoparticles with an ultrathin oxide layer. This is achieved by sequential deposition of ultrathin Al2 O3 coats, Pt, and a thick Al2 O3 layer on carbon nanocoils templates by atomic layer deposition (ALD), followed by removal of the templates. Compared with the Pt catalysts confined in Al2 O3 nanotubes without the ultrathin coats, the ultrathin coated samples have larger Pt-Al2 O3 interfaces. The maximized interfaces significantly improve the activity and the protecting Al2 O3 nanotubes retain the stability for hydrogenation reactions of 4-nitrophenol. We believe that applying ALD ultrathin coats on confined catalysts is a promising way to achieve enhanced performance for other catalysts. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Radioiodination of scorpion and snake toxins. [/sup 125/I, /sup 127/I

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

Rochat, H.; Tessier, M.; Miranda, F.

1977-10-01

Several scorpion and snake toxins were radioiodinated using the lactoperoxydase method of (/sup 125/I)iodide oxidation. Two techniques of labeling were set up: Using carrier-free Na/sup 125/I and 5 ..mu..g of toxin, about one iodine atom was incorporated per mole of protein without loss of toxicity. Specific radioactivities about 2,000 Ci/mmol (280 ..mu..Ci/..mu..g) were obtained. The modified toxin, purified by immunoprecipitation with an antiserum prepared against the native toxin, was obtained in a short time (4 hr), with a good yield (50 to 80%), and in a small volume (1 ml). Using Na/sup 127/I traced with Na /sup 125/I and largermore » amounts (200 ..mu..g) of toxin, more than one iodine atom was incorporated per mole of protein without loss of activity. Lower specific radioactivities (1 to 1.5 Ci/mmol) were obtained. The iodinated toxins were purified by gel filtration of the radioiodination mixtures on a column made of two layers of Sephadex (G-15 and G-50). The modified proteins were extensively analyzed by paper electrophoresis and polyacrylamide gel electrophoresis. Their content of monoiodotyrosine and diiodotyrosine was estimated and, in the case of toxin I of Androctonus australis Hector, it was possible to follow the iodination rate of its three tyrosine residues by automatic Edman degradation. The mode of purification of the iodinated scorpion toxins affects their behavior on molecular sieving on Sephadex G-50 and on electrophoresis on polyacrylamide gel. The results are discussed.« less

Comparison of dye doping and ultrathin emissive layer in white organic light-emitting devices with dual emissive layers

NASA Astrophysics Data System (ADS)

Wang, Xu; Qi, Yige; Yu, Junsheng

2014-09-01

White organic light-emitting devices (WOLEDs) with combined doping emissive layer (EML) and ultrathin EML have been fabricated to investigate the effect of each EML on the electroluminescent (EL) performance of the WOLEDs. Through tailoring doping concentration of bis[(4,6-difluorophenyl)-pyridinato-N,C2'](picolinate) iridium(III) (FIrpic) and thickness of ultrathin bis[2-(4-tertbutylphenyl)benzothiazolato-N,C2'] iridium (acetylacetonate) [(tbt)2Ir(acac)] EML, it is found that the change in the doping ratio of FIrpic significantly influenced the EL efficiencies and spectra, while the alteration of ultrathin EML thickness had much milder effect on the EL performance. The results indicated that ultrathin EML is in favor of reproducibility in mass production compared with doping method.

Surface passivation investigation on ultra-thin atomic layer deposited aluminum oxide layers for their potential application to form tunnel layer passivated contacts

NASA Astrophysics Data System (ADS)

Xin, Zheng; Ling, Zhi Peng; Nandakumar, Naomi; Kaur, Gurleen; Ke, Cangming; Liao, Baochen; Aberle, Armin G.; Stangl, Rolf

2017-08-01

The surface passivation performance of atomic layer deposited ultra-thin aluminium oxide layers with different thickness in the tunnel layer regime, i.e., ranging from one atomic cycle (∼0.13 nm) to 11 atomic cycles (∼1.5 nm) on n-type silicon wafers is studied. The effect of thickness and thermal activation on passivation performance is investigated with corona-voltage metrology to measure the interface defect density D it(E) and the total interface charge Q tot. Furthermore, the bonding configuration variation of the AlO x films under various post-deposition thermal activation conditions is analyzed by Fourier transform infrared spectroscopy. Additionally, poly(3,4-ethylenedioxythiophene) poly(styrene sulfonate) is used as capping layer on ultra-thin AlO x tunneling layers to further reduce the surface recombination current density to values as low as 42 fA/cm2. This work is a useful reference for using ultra-thin ALD AlO x layers as tunnel layers in order to form hole selective passivated contacts for silicon solar cells.

Simultaneous Separation of Acidic and Basic Isoperoxidases in Wounded Potato Tissue by Acrylamide Gel Electrophoresis 1

PubMed Central

Borchert, Rolf; Decedue, Charles J.

1978-01-01

Preparation and use of a newly developed pH 4.3 horizontal thin layer acrylamide gel which permits the simultaneous separation of acidic and basic isoperoxidases in up to 30 samples is described. Use of cytochrome c, horseradish peroxidase, and a purified potato isoperoxidase as internal standards for a range in isoelectric points of peroxidases from pH 3 to 11 is introduced to facilitate comparison of results obtained with different materials and different methods. Distribution of tissue-specific isoperoxidases in different cell layers of wounded potato (Solanum tuberosum L.) tissue is shown and their purification described. Evidence for the in vitro degradation of basic potato isoperoxidases resulting in more acidic forms similar to isoperoxidases occurring in wounded potato tissue is presented. The significance of this observation for the postulated differential function of different isoperoxidases is discussed. ImagesFig. 1-3 PMID:16660608

Mechanisms involved in the hydrothermal growth of ultra-thin and high aspect ratio ZnO nanowires

NASA Astrophysics Data System (ADS)

Demes, Thomas; Ternon, Céline; Morisot, Fanny; Riassetto, David; Legallais, Maxime; Roussel, Hervé; Langlet, Michel

2017-07-01

Hydrothermal synthesis of ZnO nanowires (NWs) with tailored dimensions, notably high aspect ratios (AR) and small diameters, is a major concern for a wide range of applications and still represents a challenging and recurring issue. In this work, an additive-free and reproducible hydrothermal procedure has been developed to grow ultra-thin and high AR ZnO NWs on sol-gel deposited ZnO seed layers. Controlling the substrate temperature and using a low reagent concentration (1 mM) has been found to be essential for obtaining such NWs. We show that the NW diameter remains constant at about 20-25 nm with growth time contrary to the NW length that can be selectively increased leading to NWs with ARs up to 400. On the basis of investigated experimental conditions along with thermodynamic and kinetic considerations, a ZnO NW growth mechanism has been developed which involves the formation and growth of nuclei followed by NW growth when the nuclei reach a critical size of about 20-25 nm. The low reagent concentration inhibits NW lateral growth leading to ultra-thin and high AR NWs. These NWs have been assembled into electrically conductive ZnO nanowire networks, which opens attractive perspectives toward the development of highly sensitive low-cost gas- or bio-sensors.

Probing Biomolecular Structures and Dynamics of Single Molecules Using In-Gel Alternating-Laser Excitation

PubMed Central

Santoso, Yusdi; Kapanidis, Achillefs N.

2009-01-01

Gel electrophoresis is a standard biochemical technique used for separating biomolecules on the basis of size and charge. Despite the use of gels in early single-molecule experiments, gel electrophoresis has not been widely adopted for single-molecule fluorescence spectroscopy. We present a novel method that combines gel electrophoresis and single-molecule fluorescence spectroscopy to simultaneously purify and analyze biomolecules in a gel matrix. Our method, in-gel ALEX, uses non-denaturing gels to purify biomolecular complexes of interest from free components, aggregates, and non-specific complexes. The gel matrix also slows down translational diffusion of molecules, giving rise to long, high-resolution time traces without surface immobilization, which allow extended observations of conformational dynamics in a biologically friendly environment. We demonstrated the compatibility of this method with different types of single molecule spectroscopy techniques, including confocal detection and fluorescence-correlation spectroscopy. We demonstrated that in-gel ALEX can be used to study conformational dynamics at the millisecond timescale; by studying a DNA hairpin in gels, we directly observed fluorescence fluctuations due to conformational interconversion between folded and unfolded states. Our method is amenable to the addition of small molecules that can alter the equilibrium and dynamic properties of the system. In-gel ALEX will be a versatile tool for studying structures and dynamics of complex biomolecules and their assemblies. PMID:19863108

Aligning Goals, Assessments, and Activities: An Approach to Teaching PCR and Gel Electrophoresis

PubMed Central

Robertson, Amber L.; Batzli, Janet; Harris, Michelle; Miller, Sarah

2008-01-01

Polymerase chain reaction (PCR) and gel electrophoresis have become common techniques used in undergraduate molecular and cell biology labs. Although students enjoy learning these techniques, they often cannot fully comprehend and analyze the outcomes of their experiments because of a disconnect between concepts taught in lecture and experiments done in lab. Here we report the development and implementation of novel exercises that integrate the biological concepts of DNA structure and replication with the techniques of PCR and gel electrophoresis. Learning goals were defined based on concepts taught throughout the cell biology lab course and learning objectives specific to the PCR and gel electrophoresis lab. Exercises developed to promote critical thinking and target the underlying concepts of PCR, primer design, gel analysis, and troubleshooting were incorporated into an existing lab unit based on the detection of genetically modified organisms. Evaluative assessments for each exercise were aligned with the learning goals and used to measure student learning achievements. Our analysis found that the exercises were effective in enhancing student understanding of these concepts as shown by student performance across all learning goals. The new materials were particularly helpful in acquiring relevant knowledge, fostering critical-thinking skills, and uncovering prevalent misconceptions. PMID:18316813

Electron transport in ultra-thin films and ballistic electron emission microscopy

NASA Astrophysics Data System (ADS)

Claveau, Y.; Di Matteo, S.; de Andres, P. L.; Flores, F.

2017-03-01

We have developed a calculation scheme for the elastic electron current in ultra-thin epitaxial heterostructures. Our model uses a Keldysh’s non-equilibrium Green’s function formalism and a layer-by-layer construction of the epitaxial film. Such an approach is appropriate to describe the current in a ballistic electron emission microscope (BEEM) where the metal base layer is ultra-thin and generalizes a previous one based on a decimation technique appropriated for thick slabs. This formalism allows a full quantum mechanical description of the transmission across the epitaxial heterostructure interface, including multiple scattering via the Dyson equation, which is deemed a crucial ingredient to describe interfaces of ultra-thin layers properly in the future. We introduce a theoretical formulation needed for ultra-thin layers and we compare with results obtained for thick Au(1 1 1) metal layers. An interesting effect takes place for a width of about ten layers: a BEEM current can propagate via the center of the reciprocal space (\\overlineΓ ) along the Au(1 1 1) direction. We associate this current to a coherent interference finite-width effect that cannot be found using a decimation technique. Finally, we have tested the validity of the handy semiclassical formalism to describe the BEEM current.

A comparison of non-typhoidal Salmonella from humans and food animals using pulsed-field gel electrophoresis and antimicrobial susceptibility patterns

USDA-ARS?s Scientific Manuscript database

Salmonellosis is one of the most important foodborne diseases affecting humans. To characterize the relationship between Salmonella causing human infections and their food animal reservoirs, we compared pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility patterns of non-typhoida...

Indirect fluorometric detection techniques on thin layer chromatography and effect of ultrasound on gel electrophoresis

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

Yinfa, Ma.

Thin-layer chromatography (TLC) is a broadly applicable separation technique. It offers many advantages over high performance liquid chromatography (HPLC), such as easily adapted for two-dimensional separation, for whole-column'' detection and for handling multiple samples, etc. However, due to its draggy development of detection techniques comparing with HPLC, TLC has not received the attention it deserves. Therefore, exploring new detection techniques is very important to the development of TLC. It is the principal of this dissertation to present a new detection method for TLC -- indirect fluorometric detection method. This detection technique is universal sensitive, nondestructive, and simple. This will bemore » described in detail from Sections 1 through Section 5. Section 1 and 3 describe the indirect fluorometric detection of anions and nonelectrolytes in TLC. In Section 2, a detection method for cations based on fluorescence quenching of ethidium bromide is presented. In Section 4, a simple and interesting TLC experiment is designed, three different fluorescence detection principles are used for the determination of caffeine, saccharin and sodium benzoate in beverages. A laser-based indirect fluorometric detection technique in TLC is developed in Section 5. Section 6 is totally different from Sections 1 through 5. An ultrasonic effect on the separation of DNA fragments in agarose gel electrophoresis is investigated. 262 refs.« less

Formation and prevention of fractures in sol-gel-derived thin films.

PubMed

Kappert, Emiel J; Pavlenko, Denys; Malzbender, Jürgen; Nijmeijer, Arian; Benes, Nieck E; Tsai, Peichun Amy

2015-02-07

Sol-gel-derived thin films play an important role as the functional coatings for various applications that require crack-free films to fully function. However, the fast drying process of a standard sol-gel coating often induces mechanical stresses, which may fracture the thin films. An experimental study on the crack formation in sol-gel-derived silica and organosilica ultrathin (submicron) films is presented. The relationships among the crack density, inter-crack spacing, and film thickness were investigated by combining direct micrograph analysis with spectroscopic ellipsometry. It is found that silica thin films are more prone to fracturing than organosilica films and have a critical film thickness of 300 nm, above which the film fractures. In contrast, the organosilica films can be formed without cracks in the experimentally explored regime of film thickness up to at least 1250 nm. These results confirm that ultrathin organosilica coatings are a robust silica substitute for a wide range of applications.

Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis

PubMed Central

Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

2017-01-01

Background: As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Objective: Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Materials and Methods: Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). Results: The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Conclusions: Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. SUMMARY The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines PMID:28250651

Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis.

PubMed

Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

2017-01-01

As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used : SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis : CS, TCMs: Traditional Chinese medicines.

Application of multiplex PCR, pulsed-field gel electrophoresis (PFGE), and BOX-PCR for molecular analysis of enterococci

USDA-ARS?s Scientific Manuscript database

The objective of the study was to use band-based molecular methods including BOX-PCR (Polymerase Chain Reaction) and Pulsed-Field Gel Electrophoresis (PFGE) to determine if genetically related enterococci were found among different stores, food types, or years. Enterococci were also characterized f...

Simple and Rapid System for Two-Dimensional Gel Electrophoresis Technique: A Laboratory Exercise for High School Students

ERIC Educational Resources Information Center

Maurye, Praveen; Basu, Arpita; Biswas, Jayanta Kumar; Bandyopadhyay, Tapas Kumar; Naskar, Malay

2018-01-01

Polyacrylamide gel electrophoresis (PAGE) is the most classical technique favored worldwide for resolution of macromolecules in many biochemistry laboratories due to its incessant advanced developments and wide modifications. These ever-growing advancements in the basic laboratory equipments lead to emergence of many expensive, complex, and tricky…

Analysis of Endonuclease R·EcoRI Fragments of DNA from Lambdoid Bacteriophages and Other Viruses by Agarose-Gel Electrophoresis

PubMed Central

Helling, Robert B.; Goodman, Howard M.; Boyer, Herbert W.

1974-01-01

By means of agarose-gel electrophoresis, endonuclease R·EcoRI-generated fragments of DNA from various viruses were separated, their molecular weights were determined, and complete or partial fragment maps for lambda, φ80, and hybrid phages were constructed. Images PMID:4372397

Aligning Goals, Assessments, and Activities: An Approach to Teaching PCR and Gel Electrophoresis

ERIC Educational Resources Information Center

Phillips, Allison R.; Robertson, Amber L.; Batzli, Janet; Harris, Michelle; Miller, Sarah

2008-01-01

Polymerase chain reaction (PCR) and gel electrophoresis have become common techniques used in undergraduate molecular and cell biology labs. Although students enjoy learning these techniques, they often cannot fully comprehend and analyze the outcomes of their experiments because of a disconnect between concepts taught in lecture and experiments…

Coupling Sodium Dodecyl Sulfate–Capillary Polyacrylamide Gel Electrophoresis with MALDI-TOF-MS via a PTFE Membrane

PubMed Central

Lu, Joann J.; Zhu, Zaifang; Wang, Wei; Liu, Shaorong

2011-01-01

Sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) is a fundamental analytical technique for proteomic research, and SDS–capillary gel electrophoresis (CGE) is its miniaturized version. Compared to conventional slab-gel electrophoresis, SDS-CGE has many advantages such as increased separation efficiency, reduced separation time and automated operation. SDS-CGE is not widely accepted in proteomic research primarily due to the difficulties in identifying the well-resolved proteins. MALDI–TOF–MS is an outstanding platform for protein identifications. Coupling the two would solve the problem but is extremely challenging because the MS detector has no access to the SDS-CGE resolved proteins and the SDS interferes with MS detection. In this work we introduce an approach to address these issues. We discover that poly(tetrafluoroethylene) (PTFE) membranes are excellent materials for collecting SDS-CGE separated proteins. We demonstrate that we can wash off the SDS bound to the collected proteins and identify these proteins on-membrane with MALDI-TOF-MS. We also show that we can immunoblot and Coomassie-stain the proteins collected on these membranes. PMID:21309548

Apparatus for electrophoresis separation

DOEpatents

Anderson, Norman L.

1978-01-01

An apparatus is disclosed for simultaneously performing electrophoresis separations on a plurality of slab gels containing samples of protein, protein subunits or nucleic acids. A reservoir of buffer solution is divided into three compartments by two parallel partitions having vertical slots spaced along their length. A sheet of flexible, electrically insulative material is attached to each partition and is provided with vertical slits aligned with the slots. Slab-gel holders are received within the slots with the flexible material folded outwardly as flaps from the slits to overlay portions of the holder surfaces and thereby act as electrical and liquid seals. An elongated, spaghetti-like gel containing a sample of specimen that was previously separated by isoelectric focusing techniques is vertically positioned along a marginal edge portion of the slab gel. On application of an electrical potential between the two outer chambers of buffer solution, a second dimensional electrophoresis separation in accordance with molecular weight occurs as the specimen molecules migrate across the slab gel.

Effect of processing parameters on microstructure of MoS{sub 2} ultra-thin films synthesized by chemical vapor deposition method

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

Song, Yang; You, Suping; Sun, Kewei

2015-06-15

MoS{sub 2} ultra-thin layers are synthesized using a chemical vapor deposition method based on the sulfurization of molybdenum trioxide (MoO{sub 3}). The ultra-thin layers are characterized by X-ray diffraction (XRD), photoluminescence (PL) spectroscopy and atomic force microscope (AFM). Based on our experimental results, all the processing parameters, such as the tilt angle of substrate, applied voltage, heating time and the weight of source materials have effect on the microstructures of the layers. In this paper, the effects of such processing parameters on the crystal structures and morphologies of the as-grown layers are studied. It is found that the film obtainedmore » with the tilt angle of 0.06° is more uniform. A larger applied voltage is preferred to the growth of MoS{sub 2} thin films at a certain heating time. In order to obtain the ultra-thin layers of MoS{sub 2}, the weight of 0.003 g of source materials is preferred. Under our optimal experimental conditions, the surface of the film is smooth and composed of many uniformly distributed and aggregated particles, and the ultra-thin MoS{sub 2} atomic layers (1∼10 layers) covers an area of more than 2 mm×2 mm.« less

Enhancement of absorption and color contrast in ultra-thin highly absorbing optical coatings

NASA Astrophysics Data System (ADS)

Kats, Mikhail A.; Byrnes, Steven J.; Blanchard, Romain; Kolle, Mathias; Genevet, Patrice; Aizenberg, Joanna; Capasso, Federico

2013-09-01

Recently a new class of optical interference coatings was introduced which comprises ultra-thin, highly absorbing dielectric layers on metal substrates. We show that these lossy coatings can be augmented by an additional transparent subwavelength layer. We fabricated a sample comprising a gold substrate, an ultra-thin film of germanium with a thickness gradient, and several alumina films. The experimental reflectivity spectra showed that the additional alumina layer increases the color range that can be obtained, in agreement with calculations. More generally, this transparent layer can be used to enhance optical absorption, protect against erosion, or as a transparent electrode for optoelectronic devices.

The influence of oxalate-promoted growth of saponite and talc crystals

USGS Publications Warehouse

Schumann, Dirk; Hartman, Hyman; Eberl, Dennis D.; Sears, S. Kelly; Hesse, Reinhard; Vali, Hojatollah

2013-01-01

The intercalating growth of new silicate layers or metal hydroxide layers in the interlayer space of other clay minerals is known from various mixed-layer clay minerals such as illite-smectite (I-S), chlorite-vermiculite, and mica-vermiculite. In a recent study, the present authors proposed that smectite-group minerals can be synthesized from solution as new 2:1 silicate layers within the low-charge interlayers of rectorite. That study showed how oxalate catalyzes the crystallization of saponite from a silicate gel at low temperatures (60ºC) and ambient pressure. As an extension of this work the aim of the present study was to test the claim that new 2:1 silicate layers can be synthesized as new intercalating layers in the low-charge interlayers of rectorite and whether oxalate could promote such an intercalation synthesis. Two experiments were conducted at 60ºC and atmospheric pressure. First, disodium oxalate solution was added to a suspension of rectorite in order to investigate the effects that oxalate anions have on the structure of rectorite. In a second experiment, silicate gel of saponitic composition (calculated interlayer charge −0.33 eq/O10(OH)2) was mixed with a suspension of rectorite and incubated in disodium oxalate solution. The synthesis products were extracted after 3 months and analyzed by X-ray diffraction and high-resolution transmission electron microscopy (HRTEM). The treatment of ultrathin sections with octadecylammonium (nC = 18) cations revealed the presence of 2:1 layer silicates with different interlayer charges that grew from the silicate gel. The oxalate-promoted nucleation of saponite and talc crystallites on the rectorite led to the alteration and ultimately to the destruction of the rectorite structure. The change was documented in HRTEM lattice-fringe images. The crystallization of new 2:1 layer silicates also occurred within the expandable interlayers of rectorite but not as new 2:1 silicate layers parallel to the previous 2:1 silicate layers. Instead, they grew independently of any orientation predetermined by the rectorite crystal substrate and their crystallization was responsible for the destruction of the rectorite structure.

Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis.

PubMed

Riond, Barbara; Wenger-Riggenbach, Bettina; Hofmann-Lehmann, Regina; Lutz, Hans

2009-03-01

Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and alpha-, beta-, and gamma-globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm-blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi-automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within-run and within-assay precision. Data from warm-blooded and draught horses were compared using the Mann-Whitney U test. Within-run and within-assay CVs were <5% for all protein fractions. No significant difference was found between warm-blooded and heavy draught horses and so combined reference intervals (2.5-97.5%) were calculated for total protein (51.0-72.0 g/L), albumin (29.6-38.5 g/L), alpha(1)-globulin (1.9-3.1 g/L), alpha(2)-globulin (5.3-8.7 g/L), beta(1)-globulin (2.8-7.3g/L), beta(2)-globulin (2.2-6.0 g/L), and gamma-globulin (5.8-12.7 g/L) concentrations, and albumin/globulin ratio (0.93-1.65). Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.

A multi-channel gel electrophoresis and continuous fraction collection apparatus for high throughput protein separation and characterization

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

Choi, Megan; Nordmeyer, Robert A.; Cornell, Earl

2009-10-02

To facilitate a direct interface between protein separation by PAGE and protein identification by mass spectrometry, we developed a multichannel system that continuously collects fractions as protein bands migrate off the bottom of gel electrophoresis columns. The device was constructed using several short linear gel columns, each of a different percent acrylamide, to achieve a separation power similar to that of a long gradient gel. A Counter Free-Flow elution technique then allows continuous and simultaneous fraction collection from multiple channels at low cost. We demonstrate that rapid, high-resolution separation of a complex protein mixture can be achieved on this systemmore » using SDS-PAGE. In a 2.5 h electrophoresis run, for example, each sample was separated and eluted into 48-96 fractions over a mass range of 10-150 kDa; sample recovery rates were 50percent or higher; each channel was loaded with up to 0.3 mg of protein in 0.4 mL; and a purified band was eluted in two to three fractions (200 L/fraction). Similar results were obtained when running native gel electrophoresis, but protein aggregation limited the loading capacity to about 50 g per channel and reduced resolution.« less

Visualization of DNA molecules in time during electrophoresis

NASA Technical Reports Server (NTRS)

Lubega, Seth

1991-01-01

For several years individual DNA molecules have been observed and photographed during agarose gel electrophoresis. The DNA molecule is clearly the largest molecule known. Nevertheless, the largest molecule is still too small to be seen using a microscope. A technique developed by Morikawa and Yanagida has made it possible to visualize individual DNA molecules. When these long molecules are labeled with appropriate fluorescence dyes and observed under a fluorescence microscope, although it is not possible to directly visualize the local ultrastructure of the molecules, yet because they are long light emitting chains, their microscopic dynamical behavior can be observed. This visualization works in the same principle that enables one to observe a star through a telescope because it emits light against a dark background. The dynamics of individual DNA molecules migrating through agarose matrix during electrophoresis have been described by Smith et al. (1989), Schwartz and Koval (1989), and Bustamante et al. (1990). DNA molecules during agarose gel electrophoresis advance lengthwise thorough the gel in an extended configuration. They display an extension-contraction motion and tend to bunch up in their leading ends as the 'heads' find new pores through the gel. From time to time they get hooked on obstacles in the gel to form U-shaped configurations before they resume their linear configuration.

Fundamental limits of ultrathin metasurfaces

PubMed Central

Arbabi, Amir; Faraon, Andrei

2017-01-01

We present a set of universal relations which relate the local transmission, reflection, and polarization conversion coefficients of a general class of non-magnetic passive ultrathin metasurfaces. We show that these relations are a result of equal forward and backward scattering by single layer ultrathin metasurfaces, and they lead to confinement of the transmission, reflection, and polarization conversion coefficients to limited regions of the complex plane. Using these relations, we investigate the effect of the presence of a substrate, and show that the maximum polarization conversion efficiency for a transmissive metasurface decreases as the refractive index contrast between the substrate and cladding layer increases. Furthermore, we demonstrate that a single layer reflective metasurface can achieve full 2π phase shift coverage without altering the polarization if it is illuminated from the higher refractive index material. We also discuss two approaches for achieving asymmetric scattering from metasurfaces, and realizing metasurfaces which overcome the performance limitations of single layer ultrathin metasurfaces. PMID:28262739

Recent advances in preparative electrophoresis

NASA Technical Reports Server (NTRS)

Mosher, Richard A.; Thormann, Wolfgang; Egen, Ned B.; Couasnon, Pascal; Sammons, David W.

1987-01-01

Various approaches for preparative electrophoresis, and three new instruments for preparative electrophoresis are discussed. Consideration is given to isoelectric focusing, isotachophoresis, and zone electrophoresis, three gel-based electrophoresis methods. The design, functions, and performance of the Elphor VaP 21 device of Hannig (1982), the shear-stabilized BIOSTREAM separator of Thompson (1983), and the recycling isoelectric focusing device are described.

Genetic diversity demonstrated by pulsed field gel electrophoresis of Salmonella enterica isolates obtained from diverse sources in Mexico

USDA-ARS?s Scientific Manuscript database

This study was conducted to determine the genetic diversity of Salmonella isolates recovered from a variety of sources using pulsed-field gel electrophoresis (PFGE) to assess their possible relatedness. Salmonella was isolated from ca. 52% of samples from a pepper var. Bell production system. A to...

Beverage-Agarose Gel Electrophoresis: An Inquiry-Based Laboratory Exercise with Virtual Adaptation

ERIC Educational Resources Information Center

Cunningham, Steven C.; McNear, Brad; Pearlman, Rebecca S.; Kern, Scott E.

2006-01-01

A wide range of literature and experience has shown that teaching methods that promote active learning, such as inquiry-based approaches, are more effective than those that rely on passive learning. Gel electrophoresis, one of the most common laboratory techniques in molecular biology, has a wide range of applications in the life sciences. As…

Purification of human alpha uterine protein.

PubMed

Sutcliffe, R G; Bolton, A E; Sharp, F; Nicholson, L V; MacKinnon, R

1980-03-01

Human alpha uterine protein (AUP) has been prepared from extracts of decudua by antibody affinity chromatography, DEAE Sepharose chromatography and by filtration through Sephadex G-150. This procedure yielded a protein fraction containing AUP, which was labelled with 125I by chloramine T. When analysed by SDS gel electrophoresis this radioiodinated protein fraction was found to contain predominantly a single species of protein which was precipitated by antibodies against AUP in antibody-antigen crossed electrophoresis. Rabbit anti-AUP precipitated 55-65% of the tracer in a double-antibody system. Sephadex G150 gel filtration of AUP obtained before and after affinity chromatography provided a molecular weight estimate of 50000. Since SDS gel electrophoresis revealed a polypeptide molecular weight of 23000-25000, it is suggested that AUP is a dimer.

EXAFS analysis of a human Cu,Zn SOD isoform focused using non-denaturing gel electrophoresis

NASA Astrophysics Data System (ADS)

Chevreux, Sylviane; Solari, Pier Lorenzo; Roudeau, Stéphane; Deves, Guillaume; Alliot, Isabelle; Testemale, Denis; Hazemann, Jean Louis; Ortega, Richard

2009-11-01

Isoelectric point isoforms of a metalloprotein, copper-zinc superoxide dismutase (CuZnSOD), separated on electrophoresis gels were analyzed using X-ray Absorption Spectroscopy. Mutations of this protein are involved in familial cases of amyotrophic lateral sclerosis. The toxicity of mutants could be relied to defects in the metallation state. Our purpose is to establish analytical protocols to study metallation state of protein isoforms such as those from CuZnSOD. We previously highlighted differences in the copper oxidation state between CuZnSOD isoforms using XANES. Here, we present the first results for EXAFS analyses performed at Cu and Zn K-edge on the majoritary expressed isoform of human CuZnSOD separated on electrophoresis gels.

Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

DOEpatents

Zhang, Jian-Shi; Giometti, Carol S.; Tollaksen, Sandra L.

1989-01-01

After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a DC power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, so that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. A high percentage extraction of proteins is achieved. The extracted proteins can be removed and subjected to partial digestion by trypsin or the like, followed by two-dimensional electrophoresis, resulting in a gel slab having a pattern of peptide gel spots which can be cored out and subjected to electrophoretic extraction to extract individual peptides.

High-efficiency/CRI/color stability warm white organic light-emitting diodes by incorporating ultrathin phosphorescence layers in a blue fluorescence layer

NASA Astrophysics Data System (ADS)

Miao, Yanqin; Wang, Kexiang; Zhao, Bo; Gao, Long; Tao, Peng; Liu, Xuguang; Hao, Yuying; Wang, Hua; Xu, Bingshe; Zhu, Furong

2018-01-01

By incorporating ultrathin (<0.1 nm) green, yellow, and red phosphorescence layers with different sequence arrangements in a blue fluorescence layer, four unique and simplified fluorescence/phosphorescence (F/P) hybrid, white organic light-emitting diodes (WOLEDs) were obtained. All four devices realize good warm white light emission, with high color rending index (CRI) of >80, low correlated color temperature of <3600 K, and high color stability at a wide voltage range of 5 V-9 V. These hybrid WOLEDs also reveal high forward-viewing external quantum efficiencies (EQE) of 17.82%-19.34%, which are close to the theoretical value of 20%, indicating an almost complete exciton harvesting. In addition, the electroluminescence spectra of the hybrid WOLEDs can be easily improved by only changing the incorporating sequence of the ultrathin phosphorescence layers without device efficiency loss. For example, the hybrid WOLED with an incorporation sequence of ultrathin red/yellow/green phosphorescence layers exhibits an ultra-high CRI of 96 and a high EQE of 19.34%. To the best of our knowledge, this is the first WOLED with good tradeoff among device efficiency, CRI, and color stability. The introduction of ultrathin (<0.1 nm) phosphorescence layers can also greatly reduce the consumption of phosphorescent emitters as well as simplify device structures and fabrication process, thus leading to low cost. Such a finding is very meaningful for the potential commercialization of hybrid WOLEDs.

Electrophoresis Gel Quantification with a Flatbed Scanner and Versatile Lighting from a Screen Scavenged from a Liquid Crystal Display (LCD) Monitor

ERIC Educational Resources Information Center

Yeung, Brendan; Ng, Tuck Wah; Tan, Han Yen; Liew, Oi Wah

2012-01-01

The use of different types of stains in the quantification of proteins separated on gels using electrophoresis offers the capability of deriving good outcomes in terms of linear dynamic range, sensitivity, and compatibility with specific proteins. An inexpensive, simple, and versatile lighting system based on liquid crystal display backlighting is…

Rapid pulsed-field gel electrophoresis protocol for subtyping of Streptococcus suis serotype 2.

PubMed

Luey, Cindy K Y; Chu, Yiu Wai; Cheung, Terence K M; Law, Catherine C P; Chu, Man Yu; Cheung, Danny T L; Kam, Kai Man

2007-03-01

A rapid pulsed-field gel electrophoresis (PFGE) protocol for subtyping of Streptococcus suis serotype 2 was developed and evaluated using 27 clinical isolates from 22 epidemiologically unrelated patients. Results were matched against antibiogram, virulence genotyping and multi locus sequence typing (MLST). PFGE appeared to be the most discriminatory with numerical index of discrimination (D) equal to 0.87.

Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes

PubMed Central

Greenough, Lucia; Schermerhorn, Kelly M.; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E.; Gardner, Andrew F.

2016-01-01

Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3′-5′ exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. PMID:26365239

Direct analysis of in-gel proteins by carbon nanotubes-modified paper spray ambient mass spectrometry.

PubMed

Han, Feifei; Yang, Yuhan; Ouyang, Jin; Na, Na

2015-02-07

The in situ and direct extraction, desorption and ionization of in-gel intact proteins after electrophoresis has been achieved by carbon nanotubes (CNTs)-modified paper spray mass spectrometry at ambient conditions. Characteristics of CNTs (including larger surface area, smaller pore diameter and enhanced conductivity) were endowed to the porous filter paper substrate by uniformly dispersing the CNTs on the filter paper. Upon applying electric potential to the CNTs-modified paper, the in-gel proteins were extracted from the gel and subsequently migrated to the tip of the filter paper by electrophoresis-like behavior for paper spray ionization, which was monitored by extracted ion chronograms. The characterizations of modified filter papers and CNTs nanoparticles further confirmed the role of CNTs in in-gel protein extraction, protein migration as well as spray ionization at the paper tip. Under optimized conditions, a mixture of cytochrome c, lysozyme and myoglobin was successfully separated by native electrophoresis and subsequently analysed by the present method, showing a limit of detection of 10 ng per gel band. The present strategy offers a new pathway for the direct detection of in-gel intact proteins at ambient conditions without any pre-treatment (e.g. digestion, chemical extraction and desalting), which exhibits potential applications in top-down proteomics.

Prevalence and characterization of multi-drug resistant Salmonella Enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken

PubMed Central

Parvej, Md. Shafiullah; Nazir, K. H. M. Nazmul Hussain; Rahman, M. Bahanur; Jahan, Mueena; Khan, Mohammad Ferdousur Rahman; Rahman, Marzia

2016-01-01

Aim: Salmonella is an important zoonotic pathogen responsible for animal and human diseases. The aim of the present study was to determine the prevalence and stereotyping of Salmonella isolates isolated from apparently healthy poultry. Furthermore, the clonal relatedness among the isolated Salmonella serovars was assessed. Materials and Methods: A total of 150 cloacal swab samples from apparently healthy chickens were collected, and were subjected for the isolation and identification of associated Salmonella organisms. The isolated colonies were identified and characterized on the basis of morphology, cultural characters, biochemical tests, slide agglutination test, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE). Antibiotic sensitivity patterns were also investigated using commonly used antibiotics. Results: Of the 150 samples, 11 (7.33%) produced characteristics pink colony with black center on XLD agar medium, and all were culturally and biochemically confirmed to be Salmonella. All possessed serovar-specific gene SpeF and reacted uniformly with group D antisera, suggesting that all of the isolates were Salmonella Enterica serovar Gallinarum, biovar Pullorum and/or Gallinarum. Antimicrobial susceptibility testing revealed that 54.54% of the isolated Salmonella Enterica serovars were highly sensitive to ciprofloxacin, whereas the 81.81% isolates were resistant to amoxycillin, doxycycline, kanamycin, gentamycin, and tetracycline. Pulsed-field gel electrophoresis of the XbaI-digested genomic DNA exhibited identical banding patterns, suggesting that the multidrug resistant Salmonella Enterica serovars occurring in commercial layers are highly clonal in Bangladesh. Conclusion: The present study was conducted to find out the prevalence of poultry Salmonella in layer chicken and to find out the clonal relationship among them. The data in this study suggest the prevalence of Salmonella Enterica, which is multidrug resistant and highly clonal for commercial layers of Bangladesh. PMID:27051187

Room-temperature synthesis of two-dimensional ultrathin gold nanowire parallel array with tunable spacing.

PubMed

Morita, Clara; Tanuma, Hiromitsu; Kawai, Chika; Ito, Yuki; Imura, Yoshiro; Kawai, Takeshi

2013-02-05

A series of long-chain amidoamine derivatives with different alkyl chain lengths (CnAA where n is 12, 14, 16, or 18) were synthesized and studied with regard to their ability to form organogels and to act as soft templates for the production of Au nanomaterials. These compounds were found to self-assemble into lamellar structures and exhibited gelation ability in some apolar solvents. The gelation concentration, gel-sol phase transition temperature, and lattice spacing of the lamellar structures in organic solvent all varied on the basis of the alkyl chain length of the particular CnAA compound employed. The potential for these molecules to function as templates was evaluated through the synthesis of Au nanowires (NWs) in their organogels. Ultrathin Au NWs were obtained from all CnAA/toluene gel systems, each within an optimal temperature range. Interestingly, in the case of C12AA and C14AA, it was possible to fabricate ultrathin Au NWs at room temperature. In addition, two-dimensional parallel arrays of ultrathin Au NWs were self-assembled onto TEM copper grids as a result of the drying of dispersion solutions of these NWs. The use of CnAA compounds with differing alkyl chain lengths enabled precise tuning of the distance between the Au NWs in these arrays.

Optimization of the first dimension for separation by two-dimensional gel electrophoresis of basic proteins from human brain tissue.

PubMed

Pennington, Kyla; McGregor, Emma; Beasley, Clare L; Everall, Ian; Cotter, David; Dunn, Michael J

2004-01-01

A major cause of poor resolution in the alkaline pH range of two-dimensional electrophoresis (2-DE) gels is unsatisfactory separation of basic proteins in the first dimension. We have compared methods for the separation of basic proteins in the isoelectric focusing dimension of human brain proteins. The combined use of anodic cup-loading and the hydroxyethyldisulphide containing solution (DeStreak) produced better resolution in both analytical and micropreparative protein loaded 2-DE gels than the other methods investigated.

Increase in local protein concentration by field-inversion gel electrophoresis.

PubMed

Tsai, Henghang; Low, Teck Yew; Freeby, Steve; Paulus, Aran; Ramnarayanan, Kalpana; Cheng, Chung-Pui Paul; Leung, Hon-Chiu Eastwood

2007-09-26

Proteins that migrate through cross-linked polyacrylamide gels (PAGs) under the influence of a constant electric field experience negative factors, such as diffusion and non-specific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein separation efficiency. Enhancement of protein separation efficiency was investigated by implementing pulsed field-inversion gel electrophoresis (FIGE). Separation of model protein species and large protein complexes was compared between FIGE and constant field electrophoresis (CFE) in different percentages of PAGs. Band intensities of proteins in FIGE with appropriate ratios of forward and backward pulse times were superior to CFE despite longer running times. These results revealed an increase in band intensity per defined gel volume. A biphasic protein relative mobility shift was observed in percentages of PAGs up to 14%. However, the effect of FIGE on protein separation was stochastic at higher PAG percentage. Rat liver lysates subjected to FIGE in the second-dimension separation of two-dimensional polyarcylamide gel electrophoresis (2D PAGE) showed a 20% increase in the number of discernible spots compared with CFE. Nine common spots from both FIGE and CFE were selected for peptide sequencing by mass spectrometry (MS), which revealed higher final ion scores of all nine protein spots from FIGE. Native protein complexes ranging from 800 kDa to larger than 2000 kDa became apparent using FIGE compared with CFE. The present investigation suggests that FIGE under appropriate conditions improves protein separation efficiency during PAGE as a result of increased local protein concentration. FIGE can be implemented with minimal additional instrumentation in any laboratory setting. Despite the tradeoff of longer running times, FIGE can be a powerful protein separation tool.

Micropreparative capillary gel electrophoresis of DNA: rapid expressed sequence tag library construction.

PubMed

Shi, Liang; Khandurina, Julia; Ronai, Zsolt; Li, Bi-Yu; Kwan, Wai King; Wang, Xun; Guttman, András

2003-01-01

A capillary gel electrophoresis based automated DNA fraction collection technique was developed to support a novel DNA fragment-pooling strategy for expressed sequence tag (EST) library construction. The cDNA population is first cleaved by BsaJ I and EcoR I restriction enzymes, and then subpooled by selective ligation with specific adapters followed by polymerase chain reaction (PCR) amplification and labeling. Combination of this cDNA fingerprinting method with high-resolution capillary gel electrophoresis separation and precise fractionation of individual cDNA transcript representatives avoids redundant fragment selection and concomitant repetitive sequencing of abundant transcripts. Using a computer-controlled capillary electrophoresis device the transcript representatives were separated by their size and fractions were automatically collected in every 30 s into 96-well plates. The high resolving power of the sieving matrix ensured sequencing grade separation of the DNA fragments (i.e., single-base resolution) and successful fraction collection. Performance and precision of the fraction collection procedure was validated by PCR amplification of the collected DNA fragments followed by capillary electrophoresis analysis for size and purity verification. The collected and PCR-amplified transcript representatives, ranging up to several hundred base pairs, were then sequenced to create an EST library.

Trapping and breaking of in vivo nicked DNA during pulsed-field gel electrophoresis

PubMed Central

Khan, Sharik R.; Kuzminov, Andrei

2013-01-01

Pulsed field gel electrophoresis (PFGE) offers a high-resolution approach to quantify chromosomal fragmentation in bacteria, measured as percent of chromosomal DNA entering the gel. The degree of separation in PFG depends upon the size of DNA, as well as various conditions of electrophoresis, such as electric field strength (FS), time of electrophoresis, switch time and buffer composition. Here we describe a new parameter, the structural integrity of the sample DNA itself, that influences its migration through PFGs. We show that sub-chromosomal fragments containing both spontaneous and DNA damage-induced nicks are prone to breakage during PFGE. Such breakage at single strand interruptions results in artefactual decrease in molecular weight of linear DNA making accurate determination of the number of double strand breaks difficult. While breakage of nicked sub-chromosomal fragments is FS-independent, some high molecular weight sub-chromosomal fragments are also trapped within wells under the standard PFGE conditions. This trapping can be minimized by lowering the field strength and increasing the time of electrophoresis. We discuss how breakage of nicked DNA may be mechanistically linked to trapping. Our results suggest how to optimize conditions for PFGE when quantifying chromosomal fragmentation induced by DNA damage. PMID:23770235

Quantification of DNA by Agarose Gel Electrophoresis and Analysis of the Topoisomers of Plasmid and M13 DNA Following Treatment with a Restriction Endonuclease or DNA Topoisomerase I

ERIC Educational Resources Information Center

Tweedie, John W.; Stowell, Kathryn M.

2005-01-01

A two-session laboratory exercise for advanced undergraduate students in biochemistry and molecular biology is described. The first session introduces students to DNA quantification by ultraviolet absorbance and agarose gel electrophoresis followed by ethidium bromide staining. The second session involves treatment of various topological forms of…

Introducing Proteomics in the Undergraduate Curriculum: A Simple 2D Gel Electrophoresis Exercise with Serum Proteins

ERIC Educational Resources Information Center

Kim, Thomas D.; Craig, Paul A.

2010-01-01

Two-dimensional gel electrophoresis (2DGE) remains an important tool in the study of biological systems by proteomics. While the use of 2DGE is commonplace in research publications, there are few instructional laboratories that address the use of 2DGE for analyzing complex protein samples. One reason for this lack is the fact that the preparation…

Automatic multiple-sample applicator and electrophoresis apparatus

NASA Technical Reports Server (NTRS)

Grunbaum, B. W. (Inventor)

1977-01-01

An apparatus for performing electrophoresis and a multiple-sample applicator is described. Electrophoresis is a physical process in which electrically charged molecules and colloidal particles, upon the application of a dc current, migrate along a gel or a membrane that is wetted with an electrolyte. A multiple-sample applicator is provided which coacts with a novel tank cover to permit an operator either to depress a single button, thus causing multiple samples to be deposited on the gel or on the membrane simultaneously, or to depress one or more sample applicators separately by means of a separate button for each applicator.

Disaggregation of adenylate cyclase during polyacrylamide-gel electrophoresis in mixtures of ionic and non-ionic detergents.

PubMed

Newby, A C; Chrambach, A

1979-02-01

1. Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] solubilized from the rat liver plasma membrane with 1% Lubrol PX and partially purified by gel filtration in buffer containing 0.01% Lubrol PX was physically characterized by polyacrylamide-gel electrophoresis. 2. The molecular radius determined for the partially purified enzyme was 4.9nm, compared with the value of 3.9nm obtained for the enzyme before gel filtration. 3. This difference, representing an approximate doubling of the molecular volume of the enzyme, implied that aggregation with itself or other proteins had occurred during partial purification. 4. Aggregation was not reversed by electrophoresis in the presence of high Lubrol concentrations. 5. Substitution of deoxycholate or N-dodecylsarcosinate for Lubrol PX either for solubilization or during electrophoresis led to poorer resolution of membrane proteins at concentrations giving greater than 70% loss of enzyme activity. 6. Partially purified adenylate cyclase was electrophoresed in the presence of mixed micelles of Lubrol PX and deoxycholate or Lubrol PX and N-dodecylsarcosinate. Different mixtures were examined simultaneously in a suitable apparatus. 7. Electrophoresis in the presence of 0.1% Lubrol plus 0.03% deoxycholate decreased the molecular radius of the cyclase to 4.0nm, with greater than 90% recovery of enzymic activity. The net charge of the enzyme was also increased, indicating ionic detergent binding. 8. With 0.1% Lubrol plus 0.03% N-dodecylsarcosinate the molecular radius was 4.3nm, recovery approx. 50% and net charge similar to that seen in Lubrol plus deoxycholate. 9. The resolution of cyclase from bulk protein, on an analytical scale, was improved in the presence of detergent mixtures, as compared with resolution in Lubrol alone. 10. The results demonstrate the usefulness of polyacrylamide-gel electrophoresis to detect and overcome aggregation problems with membrane proteins and suggest that detergent mixtures in specific ratios may be useful in the purification of adenylate cyclase and other intrinsic membrane proteins.

Capillary Zone Electrophoresis for the Analysis of Peptides: Fostering Students' Problem-Solving and Discovery Learning in an Undergraduate Laboratory Experiment

ERIC Educational Resources Information Center

Albright, Jessica C.; Beussman, Douglas J.

2017-01-01

Capillary electrophoresis is an important analytical separation method used to study a wide variety of samples, including those of biological origin. Capillary electrophoresis may be covered in the classroom, especially in advanced analytical courses, and while many students are exposed to gel electrophoresis in biology or biochemistry…

Monitoring refolding of tailspike endorhamnosidase using capillary electrophoresis-laser induced tryptophan fluorescence

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

Jensen, P.K.; Lee, Cheng S.; King, J.A.

1997-12-31

The use of capillary electrophoresis equipped with laser-induced tryptophan fluorescence detection is presented for monitoring the refolding pathway of phage P22 tailspike endorhamnosidase. Upon initiation of refolding, tailspike polypeptides rapidly fold into structured monomeric intermediates with a high content of secondary structure. These monomeric species associate to form the triple-chain defined folding intermediates, the protrimers. Conversion of the protrimer into the native, sodium dodecyl sulfate (SDS) resistant tailspike protein is the rate-limiting step in the refolding pathway. Refolding kinetics and yield measured by capillary electrophoresis are in good agreement with those obtained via native gel electrophoresis, SDS polyacrylamide gel electrophoresismore » (SDS-PAGE) and fluorescence spectrophotometry. To enhance separation resolution between protrimer and native protein in capillary electrophoresis, the use of poly(ethylene oxide) is investigated for the introduction of a sieving separation mechanism. The increased viscosity of the electrophoresis buffer may also play a role in resolution enhancement.« less

DIGE Analysis of Human Tissues.

PubMed

Gelfi, Cecilia; Capitanio, Daniele

2018-01-01

Two-dimensional difference gel electrophoresis (2-D DIGE) is an advanced and elegant gel electrophoretic analytical tool for comparative protein assessment. It is based on two-dimensional gel electrophoresis (2-DE) separation of fluorescently labeled protein extracts. The tagging procedures are designed to not interfere with the chemical properties of proteins with respect to their pI and electrophoretic mobility, once a proper labeling protocol is followed. The two-dye or three-dye systems can be adopted and their choice depends on specific applications. Furthermore, the use of an internal pooled standard makes 2-D DIGE a highly accurate quantitative method enabling multiple protein samples to be separated on the same two-dimensional gel. The image matching and cross-gel statistical analysis generates robust quantitative results making data validation by independent technologies successful.

Pulsed-field gel electrophoresis (PFGE): application in population structure studies of bovine mastitis-causing streptococci.

PubMed

Santos-Sanches, Ilda; Chambel, Lélia; Tenreiro, Rogério

2015-01-01

Pulsed-field gel electrophoresis (PFGE) separates large DNA molecules by the use of an alternating electrical field, such that greater size resolution can be obtained when compared to normal agarose gel electrophoresis. PFGE is often employed to track pathogens and is a valuable typing scheme to detect and differentiate strains. Particularly, the contour-clamped homogeneous electric field (CHEF) PFGE system is considered to be the gold standard for use in epidemiological studies of many bacterial pathogens. Here we describe a PFGE protocol that was applicable to the study of bovine streptococci, namely, Streptococcus agalactiae (group B Streptococcus, GBS), Streptococcus dysgalactiae subsp. dysgalactiae (group C Streptococcus, GCS), and Streptococcus uberis-which are relevant pathogens causing mastitis, a highly prevalent and costly disease in dairy industry due to antibiotherapy and loss in milk production.

Blood grouping based on PCR methods and agarose gel electrophoresis.

PubMed

Sell, Ana Maria; Visentainer, Jeane Eliete Laguila

2015-01-01

The study of erythrocyte antigens continues to be an intense field of research, particularly after the development of molecular testing methods. More than 300 specificities have been described by the International Society for Blood Transfusion as belonging to 33 blood group systems. The polymerase chain reaction (PCR) is a central tool for red blood cells (RBC) genotyping. PCR and agarose gel electrophoresis are low cost, easy, and versatile in vitro methods for amplifying defined target DNA (RBC polymorphic region). Multiplex-PCR, AS-PCR (Specific Allele Polymerase Chain Reaction), and RFLP-PCR (Restriction Fragment Length Polymorphism-Polymerase Chain Reaction) techniques are usually to identify RBC polymorphisms. Furthermore, it is an easy methodology to implement. This chapter describes the PCR methodology and agarose gel electrophoresis to identify the polymorphisms of the Kell, Duffy, Kidd, and MNS blood group systems.

Laser Desorption Mass Spectrometry for DNA Sequencing and Analysis

NASA Astrophysics Data System (ADS)

Chen, C. H. Winston; Taranenko, N. I.; Golovlev, V. V.; Isola, N. R.; Allman, S. L.

1998-03-01

Rapid DNA sequencing and/or analysis is critically important for biomedical research. In the past, gel electrophoresis has been the primary tool to achieve DNA analysis and sequencing. However, gel electrophoresis is a time-consuming and labor-extensive process. Recently, we have developed and used laser desorption mass spectrometry (LDMS) to achieve sequencing of ss-DNA longer than 100 nucleotides. With LDMS, we succeeded in sequencing DNA in seconds instead of hours or days required by gel electrophoresis. In addition to sequencing, we also applied LDMS for the detection of DNA probes for hybridization LDMS was also used to detect short tandem repeats for forensic applications. Clinical applications for disease diagnosis such as cystic fibrosis caused by base deletion and point mutation have also been demonstrated. Experimental details will be presented in the meeting. abstract.

Antibacterial and Barrier Properties of Gelatin Coated by Electrospun Polycaprolactone Ultrathin Fibers Containing Black Pepper Oleoresin of Interest in Active Food Biopackaging Applications

PubMed Central

Castro-Mayorga, Jinneth Lorena

2018-01-01

The present study evaluated the effect of using electrospun polycaprolactone (PCL) as a barrier coating and black pepper oleoresin (OR) as a natural extract on the morphology, thermal, mechanical, antimicrobial, oxygen, and water vapor barrier properties of solvent cast gelatin (GEL). The antimicrobial activity of the developed multilayer system obtained by the so-called electrospinning coating technique was also evaluated against Staphylococcus aureus strains for 10 days. The results showed that the multilayer system containing PCL and OR increased the thermal resistance, elongated the GEL film, and significantly diminished its permeance to water vapor. Active multilayer systems stored in hermetically closed bottles increased their antimicrobial activity after 10 days by inhibiting the growth of Staphylococcus aureus. This study demonstrates that addition of electrospun PCL ultrathin fibers and OR improved the properties of GEL films, which promoted its potential use in active food packaging applications. PMID:29597268

Synthesizing new types of ultrathin 2D metal oxide nanosheets via half-successive ion layer adsorption and reaction

NASA Astrophysics Data System (ADS)

Gao, Linjie; Li, Yaguang; Xiao, Mu; Wang, Shufang; Fu, Guangsheng; Wang, Lianzhou

2017-06-01

Two-dimensional (2D) metal oxide nanosheets have demonstrated their great potential in a broad range of applications. The existing synthesis strategies are mainly preparing 2D nanosheets from layered and specific transition metal oxides. How to prepare the other types of metal oxides as ultrathin 2D nanosheets remains unsolved, especially for metal oxides containing alkali, alkaline earth metal, and multiple metal elements. Herein, we developed a half-successive ion layer adsorption and reaction (SILAR) method, which could synthesize those types of metal oxides as ultrathin 2D nanosheets. The synthesized 2D metal oxides nanosheets are within 1 nm level thickness and 500 m2 · g-1 level surface area. This method allows us to develop many new types of ultrathin 2D metal oxides nanosheets that have never been prepared before.

Field-dependent magnetization of BiFeO 3 in ultrathin La 0.7Sr 0.3MnO 3/BiFeO 3 superlattice

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

Fitzsimmons, Michael R.; Jia, Quanxi X.; Singh, Surendra

2015-12-02

We report the observation of field-induced magnetization of BiFeO 3 (BFO) in an ultrathin La 0.7Sr 0.3MnO 3 (LSMO)/BFO superlattice using polarized neutron reflectivity (PNR). The depth dependent structure and magnetic characterization of subnano layer thick (thickness ~ 0.7 nm each) LSMO/BFO hetrostructure is carried out using X-ray reflectivity and PNR techniques. Our PNR results indicate parallel alignment of magnetization as well as enhancement in magnetic moment across LSMO/BFO interfaces. The study showed an increase in average magnetization on increasing applied magnetic field at 10K. As a result, we observed a saturation magnetization of 110 ± 15 kA/m (~0.8 μmore » B/Fe) for ultrathin BFO layer (~2 unit cell) sandwiched between ultrathin LSMO layers (~ 2 unit cell).« less

Optimal processing for gel electrophoresis images: Applying Monte Carlo Tree Search in GelApp.

PubMed

Nguyen, Phi-Vu; Ghezal, Ali; Hsueh, Ya-Chih; Boudier, Thomas; Gan, Samuel Ken-En; Lee, Hwee Kuan

2016-08-01

In biomedical research, gel band size estimation in electrophoresis analysis is a routine process. To facilitate and automate this process, numerous software have been released, notably the GelApp mobile app. However, the band detection accuracy is limited due to a band detection algorithm that cannot adapt to the variations in input images. To address this, we used the Monte Carlo Tree Search with Upper Confidence Bound (MCTS-UCB) method to efficiently search for optimal image processing pipelines for the band detection task, thereby improving the segmentation algorithm. Incorporating this into GelApp, we report a significant enhancement of gel band detection accuracy by 55.9 ± 2.0% for protein polyacrylamide gels, and 35.9 ± 2.5% for DNA SYBR green agarose gels. This implementation is a proof-of-concept in demonstrating MCTS-UCB as a strategy to optimize general image segmentation. The improved version of GelApp-GelApp 2.0-is freely available on both Google Play Store (for Android platform), and Apple App Store (for iOS platform). © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparative Testis Tissue Proteomics Using 2-Dye Versus 3-Dye DIGE Analysis.

PubMed

Holland, Ashling

2018-01-01

Comparative tissue proteomics aims to analyze alterations of the proteome in response to a stimulus. Two-dimensional difference gel electrophoresis (2D-DIGE) is a modified and advanced form of 2D gel electrophoresis. DIGE is a powerful biochemical method that compares two or three protein samples on the same analytical gel, and can be used to establish differentially expressed protein levels between healthy normal and diseased pathological tissue sample groups. Minimal DIGE labeling can be used via a 2-dye system with Cy3 and Cy5 or a 3-dye system with Cy2, Cy3, and Cy5 to fluorescently label samples with CyDye flours pre-electrophoresis. DIGE circumvents gel-to-gel variability by multiplexing samples to a single gel and through the use of a pooled internal standard for normalization. This form of quantitative high-resolution proteomics facilitates the comparative analysis and evaluation of tissue protein compositions. Comparing tissue groups under different conditions is crucially important for advancing the biomedical field by characterization of cellular processes, understanding pathophysiological development and tissue biomarker discovery. This chapter discusses 2D-DIGE as a comparative tissue proteomic technique and describes in detail the experimental steps required for comparative proteomic analysis employing both options of 2-dye and 3-dye DIGE minimal labeling.

Combination of Multiplex PCR and PCR-Denaturing Gradient Gel Electrophoresis for Monitoring Common Sourdough-Associated Lactobacillus Species

PubMed Central

Settanni, Luca; Valmorri, Sara; van Sinderen, Douwe; Suzzi, Giovanna; Paparella, Antonello; Corsetti, Aldo

2006-01-01

A combination of denaturing gradient gel electrophoresis (DGGE) and a previously described multiplex PCR approach was employed to detect sourdough lactobacilli. Primers specific for certain groups of Lactobacillus spp. were used to amplify fragments, which were analyzed by DGGE. DGGE profiles obtained from Lactobacillus type strains acted as standards to analyze lactobacilli from four regional Abruzzo (central Italy) sourdoughs. PMID:16672538

Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes.

PubMed

Greenough, Lucia; Schermerhorn, Kelly M; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E; Gardner, Andrew F

2016-01-29

Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3'-5' exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Erythrocyte membrane protein analysis by sodium dodecyl sulphate-capillary gel electrophoresis in the diagnosis of hereditary spherocytosis.

PubMed

Debaugnies, France; Cotton, Frédéric; Boutique, Charles; Gulbis, Béatrice

2011-03-01

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) is currently the reference method for detecting protein deficiencies related to hereditary spherocytosis. The aim of the study was to evaluate an automated capillary gel electrophoresis system, the Experion instrument from BioRad, for its ability to separate and quantify the erythrocyte membrane proteins. The major erythrocyte membrane proteins (actin, protein 4.2, protein 4.1, band 3, ankyrin, α- and β-spectrin) were extracted and purified from membrane ghosts by centrifugation, immunoprecipitation and electroelution. Analyses were performed using SDS-PAGE and sodium dodecyl sulphate capillary gel electrophoresis (SDS-CGE) to establish a separation profile of the total ghosts. Then, the samples from patients received for investigations of erythrocyte membrane defects were analysed. Five of the seven expected erythrocyte membrane proteins were finally separated and identified. In the 20 studied cases, taking into account the screening test results and the clinical and family histories, the SDS-CGE method allowed us to achieve the same conclusion as with SDS-PAGE, except for the patient with elliptocytosis. The new SDS-CGE method presents interesting features that could make this instrument a powerful diagnostic tool for detection of erythrocyte membrane protein abnormalities, and can be proposed as an automated alternative method to the labour intensive SDS-PAGE analysis.

SERS Taper-Fiber Nanoprobe Modified by Gold Nanoparticles Wrapped with Ultrathin Alumina Film by Atomic Layer Deposition

PubMed Central

Xu, Wenjie; Chen, Zhenyi; Chen, Na; Zhang, Heng; Liu, Shupeng; Hu, Xinmao; Wen, Jianxiang; Wang, Tingyun

2017-01-01

A taper-fiber SERS nanoprobe modified by gold nanoparticles (Au-NPs) with ultrathin alumina layers was fabricated and its ability to perform remote Raman detection was demonstrated. The taper-fiber nanoprobe (TFNP) with a nanoscale tip size under 80 nm was made by heated pulling combined with the chemical etching method. The Au-NPs were deposited on the TFNP surface with the electrostatic self-assembly technology, and then the TFNP was wrapped with ultrathin alumina layers by the atomic layer deposition (ALD) technique. The results told us that with the increasing thickness of the alumina film, the Raman signals decreased. With approximately 1 nm alumina film, the remote detection limit for R6G aqueous solution reached 10−6 mol/L. PMID:28245618

Ultrathin platinum nanowires grown on single-layered nickel hydroxide with high hydrogen evolution activity.

PubMed

Yin, Huajie; Zhao, Shenlong; Zhao, Kun; Muqsit, Abdul; Tang, Hongjie; Chang, Lin; Zhao, Huijun; Gao, Yan; Tang, Zhiyong

2015-03-02

Design and synthesis of effective electrocatalysts for hydrogen evolution reaction in alkaline environments is critical to reduce energy losses in alkaline water electrolysis. Here we report a hybrid nanomaterial comprising of one-dimensional ultrathin platinum nanowires grown on two-dimensional single-layered nickel hydroxide. Judicious surface chemistry to generate the fully exfoliated nickel hydroxide single layers is explored to be the key for controllable growth of ultrathin platinum nanowires with diameters of about 1.8 nm. Impressively, this hybrid nanomaterial exhibits superior electrocatalytic activity for hydrogen evolution reaction in alkaline solution, which outperforms currently reported catalysts, and the obviously improved catalytic stability. We believe that this work may lead towards the development of single-layered metal hydroxide-based hybrid materials for applications in catalysis and energy conversion.

Studies on lectins. XXXII. Application of affinity electrophoresis to the study of the interaction of lectins and their derivatives with sugars.

PubMed

Horejsí, V; Tichá, M; Kocourek, J

1977-09-29

Affinity electrophoresis was used to study the sugar binding heterogeneity of lectins or their derivatives. Commercial and demetallized preparations of concanavalin A could be resolved by affinity electrophoresis into three components with different affinity to immobilized sugar. Similarly the Vicia cracca lectin obtained by affinity chromatography behaved on affinity gels as a mixture of active and inactive molecular species. Affinity electrophoresis has shown that the nonhemagglutinating acetylated lentil lectin and photo-oxidized or sulfenylated pea lectin retain their sugar binding properties; dissociation constants of saccharide complexes of these derivatives are similar to those of native lectins. The presence of specific immobilized sugar in the affinity gel improved the resolution of isolectins from Dolichos biflorus and Ricinus communis seeds.

Wheat gliadin: digital imaging and database construction using a 4-band reference system of agarose isoelectric focusing patterns.

PubMed

Black, J A; Waggamon, K A

1992-01-01

An isoelectric focusing method using thin-layer agarose gel has been developed for wheat gliadin. Using flat-bed units with a third electrode, up to 72 samples per gel may be analyzed. Advantages over traditional acid polyacrylamide gel electrophoresis methodology include: faster run times, nontoxic media, and greater sample capacity. The method is suitable for fingerprinting or purity testing of wheat varieties. Using digital images captured by a flat-bed scanner, a 4-band reference system using isoelectric points was devised. Software enables separated bands to be assigned pI values based upon reference tracks. Precision of assigned isoelectric points is shown to be on the order of 0.02 pH units. Captured images may be stored in a computer database and compared to unknown patterns to enable an identification. Parameters for a match with a stored pattern may be adjusted for pI interval required for a match, and number of best matches.

Ultrathin spinel membrane-encapsulated layered lithium-rich cathode material for advanced Li-ion batteries.

PubMed

Wu, Feng; Li, Ning; Su, Yuefeng; Zhang, Linjing; Bao, Liying; Wang, Jing; Chen, Lai; Zheng, Yu; Dai, Liqin; Peng, Jingyuan; Chen, Shi

2014-06-11

Lack of high-performance cathode materials has become a technological bottleneck for the commercial development of advanced Li-ion batteries. We have proposed a biomimetic design and versatile synthesis of ultrathin spinel membrane-encapsulated layered lithium-rich cathode, a modification by nanocoating. The ultrathin spinel membrane is attributed to the superior high reversible capacity (over 290 mAh g(-1)), outstanding rate capability, and excellent cycling ability of this cathode, and even the stubborn illnesses of the layered lithium-rich cathode, such as voltage decay and thermal instability, are found to be relieved as well. This cathode is feasible to construct high-energy and high-power Li-ion batteries.

Electrophoresis gel image processing and analysis using the KODAK 1D software.

PubMed

Pizzonia, J

2001-06-01

The present article reports on the performance of the KODAK 1D Image Analysis Software for the acquisition of information from electrophoresis experiments and highlights the utility of several mathematical functions for subsequent image processing, analysis, and presentation. Digital images of Coomassie-stained polyacrylamide protein gels containing molecular weight standards and ethidium bromide stained agarose gels containing DNA mass standards are acquired using the KODAK Electrophoresis Documentation and Analysis System 290 (EDAS 290). The KODAK 1D software is used to optimize lane and band identification using features such as isomolecular weight lines. Mathematical functions for mass standard representation are presented, and two methods for estimation of unknown band mass are compared. Given the progressive transition of electrophoresis data acquisition and daily reporting in peer-reviewed journals to digital formats ranging from 8-bit systems such as EDAS 290 to more expensive 16-bit systems, the utility of algorithms such as Gaussian modeling, which can correct geometric aberrations such as clipping due to signal saturation common at lower bit depth levels, is discussed. Finally, image-processing tools that can facilitate image preparation for presentation are demonstrated.

Photopatterned free-standing polyacrylamide gels for microfluidic protein electrophoresis.

PubMed

Duncombe, Todd A; Herr, Amy E

2013-06-07

Designed for compatibility with slab-gel polyacrylamide gel electrophoresis (PAGE) reagents and instruments, we detail development of free-standing polyacrylamide gel (fsPAG) microstructures supporting electrophoretic performance rivalling that of microfluidic platforms. For the protein electrophoresis study described here, fsPAGE lanes are comprised of a sample reservoir and contiguous separation gel. No enclosed microfluidic channels are employed. The fsPAG devices (120 μm tall) are directly photopatterned atop of and covalently attached to planar polymer or glass surfaces. Leveraging the fast <1 h design-prototype-test cycle - significantly faster than mold based fabrication techniques - we optimize the fsPAG architecture to minimize injection dispersion for rapid (<1 min) and short (1 mm) protein separations. The facile fabrication and prototyping of the fsPAGE provides researchers a powerful tool for developing custom analytical assays. We highlight the utility of assay customization by fabricating a polyacrylamide gel with a spatial pore-size distribution and demonstrate the resulting enhancement in separation performance over a uniform gel. Further, we up-scale from a unit separation to an array of 96 concurrent fsPAGE assays in 10 min run time driven by one electrode pair. The fsPAG array layout matches that of a 96-well plate to facilitate integration of the planar free standing gel array with multi-channel pipettes while remaining compatible with conventional slab-gel PAGE reagents, such as staining for label-free protein detection. Notably, the entire fsPAGE workflow from fabrication, to operation, and readout uses readily available materials and instruments - making this technique highly accessible.

Remote N2 plasma treatment to deposit ultrathin high-k dielectric as tunneling contact layer for single-layer MoS2 MOSFET

NASA Astrophysics Data System (ADS)

Qian, Qingkai; Zhang, Zhaofu; Hua, Mengyuan; Wei, Jin; Lei, Jiacheng; Chen, Kevin J.

2017-12-01

Remote N2 plasma treatment is explored as a surface functionalization technique to deposit ultrathin high-k dielectric on single-layer MoS2. The ultrathin dielectric is used as a tunneling contact layer, which also serves as an interfacial layer below the gate region for fabricating top-gate MoS2 metal-oxide-semiconductor field-effect transistors (MOSFETs). The fabricated devices exhibited small hysteresis and mobility as high as 14 cm2·V-1·s-1. The contact resistance was significantly reduced, which resulted in the increase of drain current from 20 to 56 µA/µm. The contact resistance reduction can be attributed to the alleviated metal-MoS2 interface reaction and the preserved conductivity of MoS2 below the source/drain metal contact.

Distinguishing triplet energy transfer and trap-assisted recombination in multi-color organic light-emitting diode with an ultrathin phosphorescent emissive layer

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

Xue, Qin, E-mail: xueqin19851202@163.com; Liu, Shouyin; Xie, Guohua

2014-03-21

An ultrathin layer of deep-red phosphorescent emitter tris(1-phenylisoquinoline) iridium (III) (Ir(piq){sub 3}) is inserted within different positions of the electron blocking layer fac-tris (1-phenylpyrazolato-N,C{sup 2′})-iridium(III) (Ir(ppz){sub 3}) to distinguish the contribution of the emission from the triplet exciton energy transfer/diffusion from the adjacent blue phosphorescent emitter and the trap-assisted recombination from the narrow band-gap emitter itself. The charge trapping effect of the narrow band-gap deep-red emitter which forms a quantum-well-like structure also plays a role in shaping the electroluminescent characteristics of multi-color organic light-emitting diodes. By accurately controlling the position of the ultrathin sensing layer, it is considerably easy tomore » balance the white emission which is quite challenging for full-color devices with multiple emission zones. There is nearly no energy transfer detectable if 7 nm thick Ir(ppz){sub 3} is inserted between the blue phosphorescent emitter and the ultrathin red emitter.« less

Thermally reversible gels in electrophoresis. I - Matrix characterization

NASA Technical Reports Server (NTRS)

Righetti, Pier Giorgio; Snyder, Robert S.

1988-01-01

Two series of thermally reversible hydrogen-bonded gels have been characterized: (5 pct) PVA-(4 pct) PEG and (5 pct) PVA-(0.04 pct) borate gels. They both have extremely low melting points (16-17 C) and could be of potential interest for recovery of proteins after preparative electrophoresis. The PVA-borate gels can be exploited in the pH range 7-11 by progressively increasing the borate content in the pH interval 8 to 7 and concomitantly decreasing the borate levels in the pH zone 8 to 11. It is hypothesized that the low melting point of these gels is due to the fact that they are sparingly and sparsely hydrogen bonded along the PVA chain: on the average, 1 OH group out of 3 or 4 OH groups in the PVA polymer should be engaged in H-bond formation.

Problem-solving test: Southwestern blotting.

PubMed

Szeberényi, József

2014-01-01

Terms to be familiar with before you start to solve the test: Southern blotting, Western blotting, restriction endonucleases, agarose gel electrophoresis, nitrocellulose filter, molecular hybridization, polyacrylamide gel electrophoresis, proto-oncogene, c-abl, Src-homology domains, tyrosine protein kinase, nuclear localization signal, cDNA, deletion mutants, expression plasmid, transfection, RNA polymerase II, promoter, Shine-Dalgarno sequence, polyadenylation element, affinity chromatography, Northern blotting, immunoprecipitation, sodium dodecylsulfate, autoradiography, tandem repeats. Copyright © 2014 The International Union of Biochemistry and Molecular Biology.

Small-scale enzymatic digestion of glycoproteins and proteoglycans for analysis of oligosaccharides by LC-MS and FACE gel electrophoresis.

PubMed

Estrella, Ruby P; Whitelock, John M; Roubin, Rebecca H; Packer, Nicolle H; Karlsson, Niclas G

2009-01-01

Structural characterization of oligosaccharides from proteoglycans and other glycoproteins is greatly enhanced through the use of mass spectrometry and gel electrophoresis. Sample preparation for these sensitive techniques often requires enzymatic treatments to produce oligosaccharide sequences for subsequent analysis. This chapter describes several small-scale methods for in-gel, on-blot, and in-solution enzymatic digestions in preparation for graphitized carbon liquid chromatography-mass spectrometry (LC-MS) analysis, with specific applications indicated for glycosaminoglycans (GAGs) and N-linked oligosaccharides. In addition, accompanying procedures for oligosaccharide reduction by sodium borohydride, sample desalting via carbon microcolumn, desialylation by sialidase enzyme treatment, and small-scale oligosaccharide species fractionation are included. Fluorophore-assisted carbohydrate electrophoresis (FACE) is another useful method to isolate derivatized oligosaccharides. Overall, the modularity of these techniques provides ease and flexibility for use in conjunction with mass spectrometric and electrophoretic tools for glycomic research studies.

TCA precipitation and ethanol/HCl single-step purification evaluation: One-dimensional gel electrophoresis, bradford assays, spectrofluorometry and Raman spectroscopy data on HSA, Rnase, lysozyme - Mascots and Skyline data.

PubMed

Eddhif, Balkis; Guignard, Nadia; Batonneau, Yann; Clarhaut, Jonathan; Papot, Sébastien; Geffroy-Rodier, Claude; Poinot, Pauline

2018-04-01

The data presented here are related to the research paper entitled "Study of a Novel Agent for TCA Precipitated Proteins Washing - Comprehensive Insights into the Role of Ethanol/HCl on Molten Globule State by Multi-Spectroscopic Analyses" (Eddhif et al., submitted for publication) [1]. The suitability of ethanol/HCl for the washing of TCA-precipitated proteins was first investigated on standard solution of HSA, cellulase, ribonuclease and lysozyme. Recoveries were assessed by one-dimensional gel electrophoresis, Bradford assays and UPLC-HRMS. The mechanistic that triggers protein conformational changes at each purification stage was then investigated by Raman spectroscopy and spectrofluorometry. Finally, the efficiency of the method was evaluated on three different complex samples (mouse liver, river biofilm, loamy soil surface). Proteins profiling was assessed by gel electrophoresis and by UPLC-HRMS.

Ultrathin and Atomically Flat Transition-Metal Oxide: Promising Building Blocks for Metal-Insulator Electronics.

PubMed

Cui, Qingsong; Sakhdari, Maryam; Chamlagain, Bhim; Chuang, Hsun-Jen; Liu, Yi; Cheng, Mark Ming-Cheng; Zhou, Zhixian; Chen, Pai-Yen

2016-12-21

We present a new and viable template-assisted thermal synthesis method for preparing amorphous ultrathin transition-metal oxides (TMOs) such as TiO 2 and Ta 2 O 5 , which are converted from crystalline two-dimensional (2D) transition-metal dichalcogenides (TMDs) down to a few atomic layers. X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and scanning transmission electron microscopy (STEM) were used to characterize the chemical composition and bonding, surface morphology, and atomic structure of these ultrathin amorphous materials to validate the effectiveness of our synthesis approach. Furthermore, we have fabricated metal-insulator-metal (MIM) diodes using the TiO 2 and Ta 2 O 5 as ultrathin insulating layers with low potential barrier heights. Our MIM diodes show a clear transition from direct tunneling to Fowler-Nordheim tunneling, which was not observed in previously reported MIM diodes with TiO 2 or Ta 2 O 5 as the insulating layer. We attribute the improved performance of our MIM diodes to the excellent flatness and low pinhole/defect densities in our TMO insulting layers converted from 2D TMDs, which enable the low-threshold and controllable electron tunneling transport. We envision that it is possible to use the ultrathin TMOs converted from 2D TMDs as the insulating layer of a wide variety of metal-insulator and field-effect electronic devices for various applications ranging from microwave mixing, parametric conversion, infrared photodetection, emissive energy harvesting, to ultrafast electronic switching.

DNA migration mechanism analyses for applications in capillary and microchip electrophoresis

PubMed Central

Forster, Ryan E.; Hert, Daniel G.; Chiesl, Thomas N.; Fredlake, Christopher P.; Barron, Annelise E.

2009-01-01

In 2009, electrophoretically driven DNA separations in slab gels and capillaries have the sepia tones of an old-fashioned technology in the eyes of many, even while they remain ubiquitously used, fill a unique niche, and arguably have yet to reach their full potential. For comic relief, what is old becomes new again: agarose slab gel separations are used to prepare DNA samples for “next-gen” sequencing platforms (e.g., the Illumina and 454 machines)—dsDNA molecules within a certain size range are “cut out” of a gel and recovered for subsequent “massively parallel” pyrosequencing. In this review, we give a Barron lab perspective on how our comprehension of DNA migration mechanisms in electrophoresis has evolved, since the first reports of DNA separations by CE (∼1989) until now, 20 years later. Fused silica capillaries, and borosilicate glass and plastic microchips, quietly offer increasing capacities for fast (and even “ultra-fast”), efficient DNA separations. While the channel-by-channel scaling of both old and new electrophoresis platforms provides key flexibility, it requires each unique DNA sample to be prepared in its own micro- or nanovolume. This Achille's heel of electrophoresis technologies left an opening through which pooled-sample, next-gen DNA sequencing technologies rushed. We shall see, over time, whether sharpening understanding of transitions in DNA migration modes in crosslinked gels, nanogel solutions, and uncrosslinked polymer solutions will allow electrophoretic DNA analysis technologies to flower again. Microchannel electrophoresis, after a quiet period of metamorphosis, may emerge sleeker and more powerful, to claim its own important niche applications. PMID:19582705

Selective coherent perfect absorption of subradiant mode in ultrathin bi-layer metamaterials via antisymmetric excitation

NASA Astrophysics Data System (ADS)

Tan, Wei; Zhang, Caihong; Li, Chun; Zhou, Xiaoying; Jia, Xiaoqing; Feng, Zheng; Su, Juan; Jin, Biaobing

2017-05-01

We demonstrate that the subradiant mode in ultrathin bi-layer metamaterials can be exclusively excited under two-antisymmetric-beam illumination (or equivalently, at a node of the standing wave field), while the superradiant mode is fully suppressed due to their different mode symmetry. Coherent perfect absorption (CPA) with the Lorentzian lineshape can be achieved corresponding to the subradiant mode. A theoretical model is established to distinguish the different behaviors of these two modes and to elucidate the CPA condition. Terahertz ultrathin bi-layer metamaterials on flexible polyimide substrates are fabricated and tested, exhibiting excellent agreement with theoretical predictions. This work provides physical insight into how to selectively excite the antisymmetric subradiant mode via coherence incidence.

Microplate array diagonal gel electrophoresis (MADGE), CpG-PCR and temporal thermal ramp-MADGE (Melt-MADGE) for single nucleotide analyses in populations.

PubMed

Day, I N; O'Dell, S D; Spanakis, E; Weavind, G P

1999-02-01

Important requirements for molecular genetic epidemiological studies are economy, sample parallelism, convenience of setup and accessibility, goals inadequately met by existent approaches. We invented microplate array diagonal gel electrophoresis (MADGE) to gain simultaneously the advantages of simple setup, 96-well microplate compatibility, horizontal electrophoresis, and the resolution of polyacrylamide. At essentially no equipment cost (one simple plastic gel former), 10-100-fold savings on time for sample coding, liquid transfers, and data documentation, in addition to volume reductions and gel re-use, can be achieved. MADGE is compatible with ARMS, restriction analysis and other pattern analyses. CpG-PCR is a general PCR approach to CpG sites (10-20% of all human single base variation): both primers have 3' T, and are abutted to the CpG, forcing a TaqI restriction site if the CpG is intact. Typically, a 52 bp PCR product is then cut in half. CpG-PCR also illustrates that PAGE-MADGE readily permits analysis of 'ultrashort' PCRs. Melt-MADGE employs real-time-variable-temperature electrophoresis to examine duplex mobility during melting, achieving DGGE-like de novo, mutation scanning, but with the conveniences of arbitrary programmability, MADGE compatibility and short run time. This suite of methods enhances our capability to type or scan thousands of samples simultaneously, by 10-100-fold.

Designable ultra-smooth ultra-thin solid-electrolyte interphases of three alkali metal anodes.

PubMed

Gu, Yu; Wang, Wei-Wei; Li, Yi-Juan; Wu, Qi-Hui; Tang, Shuai; Yan, Jia-Wei; Zheng, Ming-Sen; Wu, De-Yin; Fan, Chun-Hai; Hu, Wei-Qiang; Chen, Zhao-Bin; Fang, Yuan; Zhang, Qing-Hong; Dong, Quan-Feng; Mao, Bing-Wei

2018-04-09

Dendrite growth of alkali metal anodes limited their lifetime for charge/discharge cycling. Here, we report near-perfect anodes of lithium, sodium, and potassium metals achieved by electrochemical polishing, which removes microscopic defects and creates ultra-smooth ultra-thin solid-electrolyte interphase layers at metal surfaces for providing a homogeneous environment. Precise characterizations by AFM force probing with corroborative in-depth XPS profile analysis reveal that the ultra-smooth ultra-thin solid-electrolyte interphase can be designed to have alternating inorganic-rich and organic-rich/mixed multi-layered structure, which offers mechanical property of coupled rigidity and elasticity. The polished metal anodes exhibit significantly enhanced cycling stability, specifically the lithium anodes can cycle for over 200 times at a real current density of 2 mA cm -2 with 100% depth of discharge. Our work illustrates that an ultra-smooth ultra-thin solid-electrolyte interphase may be robust enough to suppress dendrite growth and thus serve as an initial layer for further improved protection of alkali metal anodes.

Disaggregation of adenylate cyclase during polyacrylamide-gel electrophoresis in mixtures of ionic and non-ionic detergents

PubMed Central

Newby, Andrew C.; Chrambach, Andreas

1979-01-01

1. Adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] solubilized from the rat liver plasma membrane with 1% Lubrol PX and partially purified by gel filtration in buffer containing 0.01% Lubrol PX was physically characterized by polyacrylamide-gel electrophoresis. 2. The molecular radius determined for the partially purified enzyme was 4.9nm, compared with the value of 3.9nm obtained for the enzyme before gel filtration. 3. This difference, representing an approximate doubling of the molecular volume of the enzyme, implied that aggregation with itself or other proteins had occurred during partial purification. 4. Aggregation was not reversed by electrophoresis in the presence of high Lubrol concentrations. 5. Substitution of deoxycholate or N-dodecylsarcosinate for Lubrol PX either for solubilization or during electrophoresis led to poorer resolution of membrane proteins at concentrations giving greater than 70% loss of enzyme activity. 6. Partially purified adenylate cyclase was electrophoresed in the presence of mixed micelles of Lubrol PX and deoxycholate or Lubrol PX and N-dodecylsarcosinate. Different mixtures were examined simultaneously in a suitable apparatus. 7. Electrophoresis in the presence of 0.1% Lubrol plus 0.03% deoxycholate decreased the molecular radius of the cyclase to 4.0nm, with greater than 90% recovery of enzymic activity. The net charge of the enzyme was also increased, indicating ionic detergent binding. 8. With 0.1% Lubrol plus 0.03% N-dodecylsarcosinate the molecular radius was 4.3nm, recovery approx. 50% and net charge similar to that seen in Lubrol plus deoxycholate. 9. The resolution of cyclase from bulk protein, on an analytical scale, was improved in the presence of detergent mixtures, as compared with resolution in Lubrol alone. 10. The results demonstrate the usefulness of polyacrylamide-gel electrophoresis to detect and overcome aggregation problems with membrane proteins and suggest that detergent mixtures in specific ratios may be useful in the purification of adenylate cyclase and other intrinsic membrane proteins. ImagesFig. 3. PMID:435255

Microchannel gel electrophoretic separation systems and methods for preparing and using

DOEpatents

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

2015-02-24

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

Microchannel gel electrophoretic separation systems and methods for preparing and using

DOEpatents

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

2013-09-03

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

Self-surface charge exfoliation and electrostatically coordinated 2D hetero-layered hybrids

PubMed Central

Yang, Min-Quan; Xu, Yi-Jun; Lu, Wanheng; Zeng, Kaiyang; Zhu, Hai; Xu, Qing-Hua; Ho, Ghim Wei

2017-01-01

At present, the technological groundwork of atomically thin two-dimensional (2D) hetero-layered structures realized by successive thin film epitaxial growth is in principle constrained by lattice matching prerequisite as well as low yield and expensive production. Here, we artificially coordinate ultrathin 2D hetero-layered metal chalcogenides via a highly scalable self-surface charge exfoliation and electrostatic coupling approach. Specifically, bulk metal chalcogenides are spontaneously exfoliated into ultrathin layers in a surfactant/intercalator-free medium, followed by unconstrained electrostatic coupling with a dissimilar transition metal dichalcogenide, MoSe2, into scalable hetero-layered hybrids. Accordingly, surface and interfacial-dominated photocatalysis reactivity is used as an ideal testbed to verify the reliability of diverse 2D ultrathin hetero-layered materials that reveal high visible-light photoreactivity, efficient charge transfer and intimate contact interface for stable cycling and storage purposes. Such a synthetic approach renders independent thickness and composition control anticipated to advance the development of ‘design-and-build' 2D layered heterojunctions for large-scale exploration and applications. PMID:28146147

Layered ultra-thin coherent structures used as electrical resistors having low-temperature coefficient of resistivity

DOEpatents

Werner, T.R.; Falco, C.M.; Schuller, I.K.

1982-08-31

A thin film resistor having a controlled temperature coefficient of resistance (TCR) ranging from negative to positive degrees kelvin and having relatively high resistivity. The resistor is a multilayer superlattice crystal containing a plurality of alternating, ultra-thin layers of two different metals. TCR is varied by controlling the thickness of the individual layers. The resistor can be readily prepared by methods compatible with thin film circuitry manufacturing techniques.

Characterization of Proteinuria in Dogue de Bordeaux Dogs, a Breed Predisposed to a Familial Glomerulonephropathy: A Retrospective Study.

PubMed

Lavoué, Rachel; Trumel, Catherine; Smets, Pascale M Y; Braun, Jean-Pierre; Aresu, Luca; Daminet, Sylvie; Concordet, Didier; Palanché, Florence; Peeters, Dominique

2015-01-01

Dogue de Bordeaux dog has been reported to be predisposed to a familial glomerulonephropathy that displays some morphological modifications reported in focal and segmental glomerulosclerosis. Prevalence of quantitatively abnormal renal proteinuria was recently reported to be 33% in this breed. The nature of the proteinuria was assessed by sodium dodecyl sulfate-agarose gel electrophoresis and determinations of urinary markers (urinary retinol-binding protein, urinary N-acetyl-β-glucosaminidase, urinary albumin and urinary immunoglobulin G) on stored specimens. Diagnostic performances of sodium dodecyl sulfate-agarose gel electrophoresis to identify dogs with elevated urinary biomarkers were assessed. Samples from 102 adult Dogue de Bordeaux dogs (47 non-proteinuric [urine protein-to-creatinine ratio ≤ 0.2], 20 borderline-proteinuric [0.2< urine protein-to-creatinine ratio ≤ 0.5] and 35 proteinuric dogs [urine protein-to-creatinine ratio >0.5]) were used, of which 2 were suffering from familial glomerulonephropathy. The electrophoretic protein patterns, for all but one proteinuric dog, were indicative of a glomerular origin and, in all dogs, the urinary albumin concentration related to creatinine concentration and the urinary immunoglobulin G concentration related to creatinine concentration were above the upper limit of the reference interval established for the breed. Sensitivity and specificity of sodium dodecyl sulfate-agarose gel electrophoresis identifying dogs with elevated urinary albumin concentration were 94% and 92%, respectively, while diagnostic performance of sodium dodecyl sulfate-agarose gel electrophoresis in detecting dogs with elevated urinary immunoglobulin G concentration yielded sensitivity and specificity of 90% and 74%, respectively. These results suggest that all proteinuric and some borderline-proteinuric Dogue de Bordeaux dogs likely have underlying glomerular lesions and that sodium dodecyl sulfate-agarose gel electrophoresis and urinary markers might be useful to screen dogs with borderline-proteinuria. Additional investigations are warranted to assess if these findings are related to the familial glomerulonephropathy.

Tuning of in-plane optical anisotropy by inserting ultra-thin InAs layer at interfaces in (001)-grown GaAs/AlGaAs quantum wells

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

Yu, J. L., E-mail: jlyu@semi.ac.cn; Key Laboratory of Semiconductor Materials Science, Institute of Semiconductors, Chinese Academy of Sciences, P.O. Box 912, Beijing 100083; Key Laboratory of Optoelectronic Materials Chemistry and Physics, Chinese Academy of Sciences, Fuzhou 350002

2015-01-07

The in-plane optical anisotropy (IPOA) in (001)-grown GaAs/AlGaAs quantum wells (QWs) with different well widths varying from 2 nm to 8 nm has been studied by reflectance difference spectroscopy. Ultra-thin InAs layers with thickness ranging from 0.5 monolayer (ML) to 1.5 ML have been inserted at GaAs/AlGaAs interfaces to tune the asymmetry in the QWs. It is demonstrated that the IPOA can be accurately tailored by the thickness of the inserted ultra-thin InAs layer at the interfaces. Strain-induced IPOA has also been extracted by using a stress apparatus. We find that the intensity of the strain-induced IPOA decreases with the thickness ofmore » the inserted InAs layer, while that of the interface-induced IPOA increases with the thickness of the InAs layer. Theoretical calculations based on 6 band k ⋅ p theory have been carried out, and good agreements with experimental results are obtained. Our results demonstrate that, the IPOA of the QWs can be greatly and effectively tuned by inserting an ultra-thin InAs layer with different thicknesses at the interfaces of QWs, which does not significantly influence the transition energies and the transition probability of QWs.« less

Optimization of Large Gel 2D Electrophoresis for Proteomic Studies of Skeletal Muscle

PubMed Central

Reed, Patrick W.; Densmore, Allison; Bloch, Robert J.

2013-01-01

We describe improved methods for large format, 2-dimensional gel electrophoresis (2-DE) that improve protein solubility and recovery, minimize proteolysis, and reduce the loss of resolution due to contaminants and manipulations of the gels, and thus enhance quantitative analysis of protein spots. Key modifications are: (i) the use of 7M urea + 2 M thiourea, instead of 9M urea, in sample preparation and in the tops of the gel tubes; (ii) standardized deionization of all solutions containing urea with a mixed bed ion exchange resin and removal of urea from the electrode solutions; and (iii) use of a new gel tank and cooling device that eliminate the need to run two separating gels in the SDS dimension. These changes make 2D-GE analysis more reproducible and sensitive, with minimal artifacts. Application of this method to the soluble fraction of muscle tissues reliably resolves ~1800 protein spots in adult human skeletal muscle and over 2800 spots in myotubes. PMID:22589104

Polygalacturonase from Rhizopus stolonifer, an Elicitor of Casbene Synthetase Activity in Castor Bean (Ricinus communis L.) Seedlings 1

PubMed Central

Lee, Sung-Chul; West, Charles A.

1981-01-01

Apparently homogeneous polygalacturonase-elicitor purified from the filtrates of Rhizopus stolonifer cultures stimulates germinating castor bean seedlings to produce greatly increased levels of casbene synthetase activity. The purification procedure involved gel-filtration chromatography on Sephadex G-25 and G-75 columns followed by cation-exchange chromatography on a Sephadex CM C-50 column. Homogeneity of the purified preparation was indicated by the results of cationic polyacrylamide disc gel electrophoresis and isoelectric focusing (pI = 8.0). The identity of the casbene elicitor activity and polygalacturonase were indicated by the coincidence of the two activities at all stages of purification, the coincidence of both activities with the single protein-staining band detected on a cationic polyacrylamide disc gel and an isoelectric focusing gel, and the identical behavior of both activities on an agarose gel affinity column. The purified polygalacturonase-elicitor is a glycoprotein with approximately 20% carbohydrate content and an estimated molecular weight of 32,000 by polyacrylamide disc gel electrophoresis. PMID:16661728

Charge transfer from an adsorbed ruthenium-based photosensitizer through an ultra-thin aluminium oxide layer and into a metallic substrate

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

Gibson, Andrew J.; Temperton, Robert H.; Handrup, Karsten

2014-06-21

The interaction of the dye molecule N3 (cis-bis(isothiocyanato)bis(2,2-bipyridyl-4,4′-dicarbo-xylato) -ruthenium(II)) with the ultra-thin oxide layer on a AlNi(110) substrate, has been studied using synchrotron radiation based photoelectron spectroscopy, resonant photoemission spectroscopy, and near edge X-ray absorption fine structure spectroscopy. Calibrated X-ray absorption and valence band spectra of the monolayer and multilayer coverages reveal that charge transfer is possible from the molecule to the AlNi(110) substrate via tunnelling through the ultra-thin oxide layer and into the conduction band edge of the substrate. This charge transfer mechanism is possible from the LUMO+2 and 3 in the excited state but not from the LUMO,more » therefore enabling core-hole clock analysis, which gives an upper limit of 6.0 ± 2.5 fs for the transfer time. This indicates that ultra-thin oxide layers are a viable material for use in dye-sensitized solar cells, which may lead to reduced recombination effects and improved efficiencies of future devices.« less

The use of field-inversion gel electrophoresis for deletion detection in Duchenne muscular dystrophy.

PubMed Central

Chen, J D; Denton, M J; Morgan, G; Pearn, J H; Mackinlay, A G

1988-01-01

Deletion is a common cause of Duchenne muscular dystrophy (DMD). Field-inversion gel electrophoresis, in conjunction with Southern blot hybridization, was used to detect large SfiI DNA fragments in the DMD locus. Two unrelated boys with DMD were found to have abnormal sized DNA fragments resulting from deletions. Some of the female relatives of these patients were also shown by this method to have deletions in the DMD locus. Images Figure 1 PMID:3358426

Case study of the use of pulsed field gel electrophoresis in the detection of a food-borne outbreak.

PubMed

De Lappe, Niall; Cormican, Martin

2015-01-01

In early July 2008, a cluster of six Salmonella Agona was identified in the Republic of Ireland. A dispersed, common source outbreak was suspected. Later in July a further case was identified and the Health Protection Agency in the UK indicated that they had 32 cases of S. Agona since Feb 2008. This chapter discusses how pulsed field gel electrophoresis was used to help confirm an outbreak and to trace the source of the outbreak.

Identification of Methanococcus Jannaschii Proteins in 2-D Gel Electrophoresis Patterns by Mass Spectrometry

DOE R&D Accomplishments Database

Liang, X.

1998-06-10

The genome of Methanococcus jannaschii has been sequenced completely and has been found to contain approximately 1,770 predicted protein-coding regions. When these coding regions are expressed and how their expression is regulated, however, remain open questions. In this work, mass spectrometry was combined with two-dimensional gel electrophoresis to identify which proteins the genes produce under different growth conditions, and thus investigate the regulation of genes responsible for functions characteristic of this thermophilic representative of the methanogenic Archaea.

Preparative electrophoresis with on-column optical fiber monitoring and direct elution into a minimized volume.

PubMed

Jackson, George W; Willson, Richard

2005-11-01

A "column-format" preparative electrophoresis device which obviates the need for gel extraction or secondary electro-elution steps is described. Separated biomolecules are continuously detected and eluted directly into a minimal volume of free solution for subsequent use. An optical fiber allows the species of interest to be detected just prior to elution from the gel column, and a small collection volume is created by addition of an ion-exchange membrane near the end of the column.

Electrophoretic extraction of proteins from two-dimensional electrophoresis gel spots

DOEpatents

Zhang, Jian-Shi; Giometti, C.S.; Tollaksen, S.L.

1987-09-04

After two-dimensional electrophoresis of proteins or the like, resulting in a polyacrylamide gel slab having a pattern of protein gel spots thereon, an individual protein gel spot is cored out from the slab, to form a gel spot core which is placed in an extraction tube, with a dialysis membrane across the lower end of the tube. Replicate gel spots can be cored out from replicate gel slabs and placed in the extraction tube. Molten agarose gel is poured into the extraction tube where the agarose gel hardens to form an immobilizing gel, covering the gel spot cores. The upper end portion of the extraction tube is filled with a volume of buffer solution, and the upper end is closed by another dialysis membrane. Upper and lower bodies of a buffer solution are brought into contact with the upper and lower membranes and are provided with electrodes connected to the positive and negative terminals of a dc power supply, thereby producing an electrical current which flows through the upper membrane, the volume of buffer solution, the agarose, the gel spot cores and the lower membrane. The current causes the proteins to be extracted electrophoretically from the gel spot cores, so that the extracted proteins accumulate and are contained in the space between the agarose gel and the upper membrane. 8 figs.

Low-cost ultra-thin broadband terahertz beam-splitter.

PubMed

Ung, Benjamin S-Y; Fumeaux, Christophe; Lin, Hungyen; Fischer, Bernd M; Ng, Brian W-H; Abbott, Derek

2012-02-27

A low-cost terahertz beam-splitter is fabricated using ultra-thin LDPE plastic sheeting coated with a conducting silver layer. The beam splitting ratio is determined as a function of the thickness of the silver layer--thus any required splitting ratio can be printed on demand with a suitable rapid prototyping technology. The low-cost aspect is a consequence of the fact that ultra-thin LDPE sheeting is readily obtainable, known more commonly as domestic plastic wrap or cling wrap. The proposed beam-splitter has numerous advantages over float zone silicon wafers commonly used within the terahertz frequency range. These advantages include low-cost, ease of handling, ultra-thin thickness, and any required beam splitting ratio can be readily fabricated. Furthermore, as the beam-splitter is ultra-thin, it presents low loss and does not suffer from Fabry-Pérot effects. Measurements performed on manufactured prototypes with different splitting ratios demonstrate a good agreement with our theoretical model in both P and S polarizations, exhibiting nearly frequency-independent splitting ratios in the terahertz frequency range.

Agarose gel electrophoresis of joint fluid using Hyrys-Hydrasys SEBIA system as a new prognostic tool for periprosthetic osteolysisin revision arthroplasty

PubMed Central

Chiva, A

2013-01-01

Rationale. Prevention of wear-mediated osteolysis, the most common complication in total joint arthroplasty, is a great challenge for orthopedic surgery. Despite the diversity of current biomarkers of periprosthetic osteolysis (products of wear, bone turnover and inflammatory biomarkers), the major interferences and the great amount of sample necessary for analysis limit their use in clinical practice. Objective. The aim of this paper is to present three new electrophoretic methods using Hyrys-Hydrasys SEBIA system that have been used for the first time in Electrophoresis Laboratory of our hospital in the analysis of joint fluid for the prevention of periprosthetic osteolysis in revision arthroplasty. Methods and results. Analytical aspects of agarose gel electrophoresis of joint fluid proteins and lipoproteins as well as SDS-agarose gel electrophoresis of joint fluid proteins, their performances and clinical value are presented. The decreased level of albumin and increased level of alpha1 and alpha2 globulins were frequent changes detected on SEBIA electropherograms and good indicator for the presence of an inflammatory reaction generated by particle debris. In addition, a slightly increase of LDL mobility could provide good information about a high oxidative stress. Moreover, the Ig G assessed by using SDS-agarose gel electrophoresis could be a potential biomarker for an immunological reaction towards orthopedic implants. Discussion. Electrophoresis of joint fluid using Hyrys-Hydrasys SEBIA France system is a new analytical technique able to remove the most of current biomarkers disadvantages due to the determination of particular proteins (acute phase proteins, albumin, lipoproteins, and immunoglobulins) by using minimal amounts of joint fluid with minor interferences, minimal cost and rapid results. Abbreviations CTX, crosslinked C-telopeptide; IL- interleukins; Ig G, immunoglobulin G; LDL, low density lipoprotein; NTX, crosslinked N-telopeptide; PICP, procollagen I C – terminal extension peptide; SDS, sodium dodecyl sulphate PMID:24146682

Diffractive intermediate layer enables broadband light trapping for high efficiency ultrathin c-Si tandem cells

NASA Astrophysics Data System (ADS)

Li, Guijun; Ho, Jacob Y. L.; Li, He; Kwok, Hoi-Sing

2014-06-01

Light management through the intermediate reflector in the tandem cell configuration is of great practical importance for achieving high stable efficiency and also low cost production. So far, however, the intermediate reflectors employed currently are mainly focused on the light absorption enhancement of the top cell. Here, we present a diffractive intermediate layer that allows for light trapping over a broadband wavelength for the ultrathin c-Si tandem solar cell. Compared with the standard intermediate reflector, this nanoscale architectural intermediate layer results in a 35% and 21% remarkable enhancement of the light absorption in the top (400-800 nm) and bottom (800-1100 nm) cells simultaneously, and ultrathin c-Si tandem cells with impressive conversion efficiency of 13.3% are made on the glass substrate.

Ultrathin Polyaniline-based Buffer Layer for Highly Efficient Polymer Solar Cells with Wide Applicability

PubMed Central

Zhao, Wenchao; Ye, Long; Zhang, Shaoqing; Fan, Bin; Sun, Mingliang; Hou, Jianhui

2014-01-01

Interfacial buffer layers often attribute the improved device performance in organic optoelectronic device. Herein, a water-soluble hydrochloric acid doped polyanilines (HAPAN) were utilized as p-type electrode buffer layer in highly efficient polymer solar cells (PSC) based on PBDTTT-EFT and several representative polymers. The PBDTTT-EFT-based conventional PSC featuring ultrathin HAPAN (1.3 nm) delivered high PCE approximately 9%, which is one of the highest values among conventional PSC devices. Moreover, ultrathin HAPAN also exhibited wide applicability in a variety of efficient photovoltaic polymers including PBDTTT-C-T, PTB7, PBDTBDD, PBTTDPP-T, PDPP3T and P3HT. The excellent performances were originated from the high transparency, small film roughness and suitable work function. PMID:25300365

Electrophoresis of DNA in agarose gels, polyacrylamide gels and in free solution

PubMed Central

Stellwagen, Nancy C.

2009-01-01

This review describes the electrophoresis of curved and normal DNA molecules in agarose gels, polyacrylamide gels and in free solution. These studies were undertaken to clarify why curved DNA molecules migrate anomalously slowly in polyacrylamide gels but not in agarose gels. Two milestone papers are cited, in which Ferguson plots were used to estimate the effective pore size of agarose and polyacrylamide gels. Subsequent studies on the effect of the electric field on agarose and polyacrylamide gel matrices, DNA interactions with the two gel matrices, and the effect of curvature on the free solution mobility of DNA are also described. The combined results suggest that the anomalously slow mobilities observed for curved DNA molecules in polyacrylamide gels are due primarily to preferential interactions of curved DNAs with the polyacrylamide gel matrix; the restrictive pore size of the matrix is of lesser importance. In free solution, DNA mobilities increase with increasing molecular mass until leveling off at a plateau value of (3.17 ± 0.01) × 10-4 cm2/Vs in 40 mM Tris-acetate-EDTA buffer at 20°C. Curved DNA molecules migrate anomalously slowly in free solution as well as in polyacrylamide gels, explaining why the Ferguson plots of curved and normal DNAs containing the same number of base pairs extrapolate to different mobilities at zero gel concentration. PMID:19517510

[Preparation of polyacrylamide gel electrophoresis for human chorionic gonadotropin chimeric peptide 12 expressed in E. coli].

PubMed

Zou, Yong-Shui; Xu, Wan-Xiang; He, Yuan; Sun, Zhi-Da; Xue, Xiao-Lin

2002-09-01

In recent years, development of chimeric peptide (CP) immunogens is a trend in the vaccinological field. The CPs contain a B cell epitope(s) of target antigen and a promiscuous self - or foreign- T cell epitope(s). However, such constructed CPs were all expressed in prokaryotic or eukaryotic systems at lower levels. To purify the human chorionic gonadotropin (hCG) CP12 expressed in E. coli at the level of about 1% of the total cell proteins, an improved method of preparative gel polyacrylamide gel electrophoresis (PAGE) was developed. The important improvement to routine preparative PAGE involves: (1) running reversed electrophoresis by rearranging the gel- carrying plate when the bromophenol blue band arrived at 1-1.5 centimeter from the bottom of the gel; (2) making a collecting trough between the gel and a dialytic membrane that was used to isolate the upper tank buffer. About 8 fractions were collected at regular intervals of 15 minutes after bromophenol blue running out of gel. And then 0.2 ml was taken from each fraction and the protein was precipitated by sequentially adding trichloroacetic acid and acetone. Each sample was dissolved in 20 microL sample buffer and analyzed and identified by SDS-PAGE and Western blotting. As a result, the hCG CP12 expression product with 95% relative homogeneity was harvested at a 50-100 microgram level after a single-step purification of this preparative PAGE, with respect to the sample which contained 3-4 mg of cell proteins.

Investigation of Cu-, Zn- and Fe-containing human brain proteins using isotopic-enriched tracers by LA-ICP-MS and MALDI-FT-ICR-MS

NASA Astrophysics Data System (ADS)

Becker, J. Susanne; Zoriy, Miroslav; Pickhardt, Carola; Przybylski, Michael; Becker, J. Sabine

2005-04-01

Identification of metal-containing proteins and determination of Cu, Fe, Zn concentration in very small protein volumes is of increasing importance in protein research. Proteins containing metal ions were analyzed directly and simultaneously in separated protein spots in two-dimensional gels (2D gels) by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) as an element mass spectrometric technique. In order to study the formation of proteins containing Cu, Zn and Fe in a human brain sample, isotopic-enriched tracers (54Fe, 65Cu and 67Zn) were doped to two-dimensional gels of separated Alzheimer-diseased brain proteins after two-dimensional (2D) gel electrophoresis. The protein spots were screened systematically by LA-ICP-MS with respect to these metal ion intensities. 54Fe/56Fe, 65Cu/63Cu and 67Zn/64Zn isotope ratios in metal-containing proteins were measured directly by LA-ICP-MS. The isotope ratio measurements obtained by LA-ICP-MS indicate certain protein spots with a natural isotope composition of Cu, Zn and/or Fe. These proteins already contained the metal investigated in the original proteins and are stable enough to survive the reducing conditions during gel electrophoresis. On the other hand, proteins with a changed isotope ratio of metals in comparison to the isotope ratio in nature demonstrate the accumulation of tracers within the protein complexes during the tracer experiments in 2D gels. The identification of singular protein spots from Alzheimer-diseased brain separated by 2D gel electrophoresis was attempted by biopolymer mass spectrometry using MALDI-FTICR-MS after excision from the 2D gel and tryptic digestion.

Confinement of holes and electrons in blue organic light-emitting diodes with additional red emissive layers

NASA Astrophysics Data System (ADS)

Kang, Jin Sung; Yoo, Seung Il; Kim, Jin Wook; Yoon, Geum Jae; Yi, Seungjun; Kim, Woo Young

2016-02-01

We used various emissive layer (EML) structures with ultrathin red EMLs to enhance the charge carrier balance and carrier recombination rate in blue PHOLED devices. These EML materials have different energy gaps between highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energy levels. The ultrathin red EMLs, which were inserted in between the blue EMLs, effectively confined the charge carriers in EML, and increased the carrier recombination rate. The thickness of the individual EML was optimized, under 30 nm of the total thickness of EML. The blue PHOLEDs with ultrathin red EMLs achieved a luminous efficiency of 19.24 cd/A, which was 28.7% higher than those without ultrathin red EMLs, and the maximum external quantum efficiency was 11.81% at 500 cd/m2.

Salmonella enterica Pulsed-Field Gel Electrophoresis Clusters, Minnesota, USA, 2001–2007

PubMed Central

Hedberg, Craig W.; Meyer, Stephanie; Boxrud, David J.; Smith, Kirk E.

2010-01-01

We determined characteristics of Salmonella enterica pulsed-field gel electrophoresis clusters that predict their being solved (i.e., that result in identification of a confirmed outbreak). Clusters were investigated by the Minnesota Department of Health by using a dynamic iterative model. During 2001–2007, a total of 43 (12.5%) of 344 clusters were solved. Clusters of >4 isolates were more likely to be solved than clusters of 2 isolates. Clusters in which the first 3 case isolates were received at the Minnesota Department of Health within 7 days were more likely to be solved than were clusters in which the first 3 case isolates were received over a period >14 days. If resources do not permit investigation of all S. enterica pulsed-field gel electrophoresis clusters, investigation of clusters of >4 cases and clusters in which the first 3 case isolates were received at a public health laboratory within 7 days may improve outbreak investigations. PMID:21029524

Genotypic analysis of strains of mutans streptococci by pulsed-field gel electrophoresis.

PubMed

Mineyama, R; Yoshino, S; Fukushima, K

2004-01-01

The species and serotypes of various strains of S. mutans and S. sobrinus were characterized by pulsed-field gel electrophoresis after the genomic DNA from the various strains had been digested with five restriction enzymes (EcoR I, Xba I, Hind III, Sfi I and BssH II) separately. Among these restriction enzymes, BssH II was very useful for the characterization of species and serotypes and, in particular, digestion discriminated between serotypes d and g. The restriction patterns obtained from the genomic DNA of isolates isolated from children's saliva were essentially identical to those from the genomic DNA of the standard laboratory strains. Patterns of BssH II digests of the genomic DNA of 10 isolates identified as S. sobrinus were characteristic of serotype g of the standard laboratory strains. Our results indicate that digestion with BssH II and subsequence analysis by pulsed-field gel electrophoresis should be useful for the characterization of species and serotypes and for epidemiological studies of mutans streptococci.

WGA-based lectin affinity gel electrophoresis: A novel method for the detection of O-GlcNAc-modified proteins.

PubMed

Kubota, Yuji; Fujioka, Ko; Takekawa, Mutsuhiro

2017-01-01

Post-translational modification with O-linked β-N-acetylglucosamine (O-GlcNAc) occurs selectively on serine and/or threonine residues of cytoplasmic and nuclear proteins, and dynamically regulates their molecular functions. Since conventional strategies to evaluate the O-GlcNAcylation level of a specific protein require time-consuming steps, the development of a rapid and easy method for the detection and quantification of an O-GlcNAcylated protein has been a challenging issue. Here, we describe a novel method in which O-GlcNAcylated and non-O-GlcNAcylated forms of proteins are separated by lectin affinity gel electrophoresis using wheat germ agglutinin (WGA), which primarily binds to N-acetylglucosamine residues. Electrophoresis of cell lysates through a gel containing copolymerized WGA selectively induced retardation of the mobility of O-GlcNAcylated proteins, thereby allowing the simultaneous visualization of both the O-GlcNAcylated and the unmodified forms of proteins. This method is therefore useful for the quantitative detection of O-GlcNAcylated proteins.

Direct synthesis of ultrathin SOI structure by extremely low-energy oxygen implantation

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

Hoshino, Yasushi, E-mail: yhoshino@kanagawa-u.ac.jp; Yachida, Gosuke; Inoue, Kodai

2016-06-15

We performed extremely low-energy {sup 16}O{sup +} implantation at 10 keV (R{sub p} ∼ 25 nm) followed by annealing aiming at directly synthesizing an ultrathin Si layer separated by a buried SiO{sub 2} layer in Si(001) substrates, and then investigated feasible condition of recrystallization and stabilization of the superficial Si and the buried oxide layer by significantly low temperature annealing. The elemental compositions were analyzed by Rutherford backscattering (RBS) and secondary ion mass spectroscopy (SIMS). The crystallinity of the superficial Si layer was quantitatively confirmed by ananlyzing RBS-channeling spectra. Cross-sectional morphologies and atomic configurations were observed by transmission electron microscopemore » (TEM). As a result, we succeeded in directly synthesizing an ultrathin single-crystalline silicon layer with ≤20 nm thick separated by a thin buried stoichiometric SiO{sub 2} layer with ≤20 nm thick formed by extremely low-energy {sup 16}O{sup +} implantation followed by surprisingly low temperature annealing at 1050{sup ∘} C.« less

Suppressing Shuttle Effect Using Janus Cation Exchange Membrane for High-Performance Lithium-Sulfur Battery Separator.

PubMed

Li, Zhen; Han, Yu; Wei, Junhua; Wang, Wenqiang; Cao, Tiantian; Xu, Shengming; Xu, Zhenghe

2017-12-27

Suppressing the shuttle effect of polysulfide ions to obtain high durability and good electrochemical performance is of great concern in the field of lithium-sulfur batteries. To address this issue, a Janus membrane consisting of an ultrathin dense layer and a robust microporous layer is fabricated using cation exchange resin. Different from the composite membranes made from polyolefin membranes, the multiple layers of the Janus membrane in this study are synchronously generated by one step, getting rid of the additional complex coating processes. Excellent overall performance is obtained by the cooperation of multiple factors. The excellent ionic selectivity of cation exchange resin renders a great suppression of the shuttle effect, endowing the lithium-sulfur battery with high Coulombic efficiency of 92.0-99.0% (LiNO 3 -free electrolyte). The ultrathin property of a dense layer renders a low ionic resistance, resulting in 60% higher discharge capacity over the entire C-rates (versus the control sample with Celgard 2400 membrane). The robust macroporous layer supports the ultrathin layer to achieve a free-standing property, ensuring the usability of the Janus membrane.

Modification of vital wheat gluten with phosphoric acid to produce high free-solution capacity

USDA-ARS?s Scientific Manuscript database

Wheat gluten reacts with phosphoric acid to produce natural superabsorbent gels. The gel properties are defined by Fourier Transform Infra-red (FTIR) spectroscopy, 2-dimensional gel electrophoresis (2DE), and uptake of water, salt solutions, and aqueous ethanol. Temperatures above 120'C and dry cond...

A single-step simultaneous protein staining procedure for polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis.

PubMed

Pal, Jayanta K; Berwal, Sunil K; Soni, Rupali N

2012-01-01

A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS) polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water and viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining), and cost-effective.

Separation of oligopeptides, nucleobases, nucleosides and nucleotides using capillary electrophoresis/electrochromatography with sol-gel modified inner capillary wall.

PubMed

Svobodová, Jana; Kofroňová, Olga; Benada, Oldřich; Král, Vladimír; Mikšík, Ivan

2017-09-29

The aim of this article is to study the modification of an inner capillary wall with sol-gel coating (pure silica sol-gel or silica sol-gel containing porphyrin-brucine conjugate) and determine its influence on the separation process using capillary electrophoresis/electrochromatography method. After modification of the inner capillary surface the separation of analytes was performed using two different phosphate buffers (pH 2.5 and 9.0) and finally the changes in electrophoretic mobilities of various samples were calculated. To confirm that the modification of the inner capillary surface was successful, the parts of the inner surfaces of capillaries were observed using scanning electron microscopy. The analytes used as testing samples were oligopeptides, nucleosides, nucleobases and finally nucleotides. Copyright © 2017 Elsevier B.V. All rights reserved.

Identification of Fur, Aconitase, and Other Proteins Expressed by Mycobacterium tuberculosis under Conditions of Low and High Concentrations of Iron by Combined Two-Dimensional Gel Electrophoresis and Mass Spectrometry

PubMed Central

Wong, Diane K.; Lee, Bai-Yu; Horwitz, Marcus A.; Gibson, Bradford W.

1999-01-01

Iron plays a critical role in the pathophysiology of Mycobacterium tuberculosis. To gain a better understanding of iron regulation by this organism, we have used two-dimensional (2-D) gel electrophoresis, mass spectrometry, and database searching to study protein expression in M. tuberculosis under conditions of high and low iron concentration. Proteins in cellular extracts from M. tuberculosis Erdman strain grown under low-iron (1 μM) and high-iron (70 μM) conditions were separated by 2-D polyacrylamide gel electrophoresis, which allowed high-resolution separation of several hundred proteins, as visualized by Coomassie staining. The expression of at least 15 proteins was induced, and the expression of at least 12 proteins was decreased under low-iron conditions. In-gel trypsin digestion was performed on these differentially expressed proteins, and the digestion mixtures were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry to determine the molecular masses of the resulting tryptic peptides. Partial sequence data on some of the peptides were obtained by using after source decay and/or collision-induced dissociation. The fragmentation data were used to search computerized peptide mass and protein sequence databases for known proteins. Ten iron-regulated proteins were identified, including Fur and aconitase proteins, both of which are known to be regulated by iron in other bacterial systems. Our study shows that, where large protein sequence databases are available from genomic studies, the combined use of 2-D gel electrophoresis, mass spectrometry, and database searching to analyze proteins expressed under defined environmental conditions is a powerful tool for identifying expressed proteins and their physiologic relevance. PMID:9864233

Affinity in electrophoresis.

PubMed

Heegaard, Niels H H

2009-06-01

The journal Electrophoresis has greatly influenced my approaches to biomolecular affinity studies. The methods that I have chosen as my main tools to study interacting biomolecules--native gel and later capillary zone electrophoresis--have been the topic of numerous articles in Electrophoresis. Below, the role of the journal in the development and dissemination of these techniques and applications reviewed. Many exhaustive reviews on affinity electrophoresis and affinity CE have been published in the last few years and are not in any way replaced by the present deliberations that are focused on papers published by the journal.

Glycosaminoglycan blotting on nitrocellulose membranes treated with cetylpyridinium chloride after agarose-gel electrophoretic separation.

PubMed

Maccari, Francesca; Volpi, Nicola

2002-09-01

We describe a method for blotting and immobilizing several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by a cationic detergent after their separation by conventional agarose gel electrophoresis. Nitrocellulose membranes were derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of glycosaminoglycans (GAGs) were capillary-blotted after their separation in agarose gel electrophoresis in barium acetate/1,2-diaminopropane. Single purified species of variously sulfated polysaccharides were transferred onto the derivatized membranes after electrophoresis with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining) permitting about 0.1 nug threshold of detection. Nonsulfated polyanions, hyaluronic acid, a fructose-containing polysaccharide with a chondroitin backbone purified from Escherichia coli U1-41, and its defructosylated product, were also electrophoretically separated and transferred onto membranes. The limit of detection for desulfated GAGs was about 0.1-0.5 nug after irreversible or reversible staining. GAG extracts from bovine, lung and aorta, and human aorta and urine were separated by agarose gel electrophoresis and blotted on CPC-treated nitrocellulose membranes. The polysaccharide composition of these extracts was determined. The membrane stained with toluidine blue (reversible staining) was destained and the same lanes used for immunological detection or other applications. Reversible staining was also applied to recover single species of polysaccharides after electrophoretic separation of mixtures of GAGs and their transfer onto membranes. Single bands were released from the membrane with an efficiency of 70-100% for further biochemical characterization.

Detection of undeclared animal by-products in commercial canine canned foods: Comparative analyses by ELISA and PCR-RFLP coupled with slab gel electrophoresis or capillary gel electrophoresis.

PubMed

Hsieh, Ming-Kun; Shih, Pei-Yin; Wei, Chia-Fong; Vickroy, Thomas W; Chou, Chi-Chung

2016-03-30

The potential presence of undeclared animal by-products in pet foods is not subject to routine examination. Previously published methods for species-based identification of animal by-products have not been used routinely owing to inconsistent results. The present study evaluated the utility of several approaches for accurate identification of animal by-products in 11 commercial brands of canine canned foods. Canine canned foods from several countries were analysed by ELISA, PCR-RFLP coupled with slab-gel electrophoresis (SGE) and capillary gel electrophoresis (CGE) to test for evidence of by-products derived from cattle, chicken, sheep or pig. While CGE-based analysis detected all (24) animal-derived by-products that were reported for the 11 test samples, SGE and ELISA detected only 22/24 (92%) and 14/24 (58%) of labelled by-products, respectively. In addition, undeclared animal by-products were found using all three analytical approaches with CGE detecting more positives (19) than SGE (17) or ELISA (5). Significant disparities were evident between the labelled contents and the detected content of animal by-products. CGE-based testing for PCR products appears to provide greater sensitivity and accuracy than either SGE or ELISA-based methods. As testing of commercial products becomes more reliable and mainstream, manufacturers will need to develop more thorough and accurate labelling protocols. © 2015 Society of Chemical Industry.

Isolation of tissue layers in hermatypic corals by N-acetylcysteine: morphological and proteomic examinations

NASA Astrophysics Data System (ADS)

Peng, S.-E.; Luo, Y.-J.; Huang, H.-J.; Lee, I.-T.; Hou, L.-S.; Chen, W.-N. U.; Fang, L.-S.; Chen, C.-S.

2008-03-01

Corals are diploblastic in body pattern and include two tissue layers, the epidermis and gastrodermis, interconnected by an acellular matrix mesoglea. During development, cells in these tissue layers differentiate morphologically and functionally. In most hermatypic corals, the gastrodermis further develops an ability to associate with microalgae dinoflagellates. This endosymbiosis occurs inside specific host gastrodermal cells, and its mechanism still remains unclear notwithstanding decades of research. The delay in progress is partly due to the difficulty in separating the gastrodermis and its symbionts from the epidermis for detailed cellular and biochemical investigations. The present study reports a simple method to separate these two tissue layers in hermatypic corals using the reducing agent, N-acetylcysteine (NAC). Efficient tissue and proteomic isolations are demonstrated by microscopy and two-dimensional SDS polyacrylamide gel electrophoresis (2D SDS-PAGE). The NAC treatment was able to separate tissue layers without inducing protein degradation. Furthermore, the sensitivity of protein detection greatly increases in the isolated tissue layers. The application of the present technique provides future research on endosymbiosis and coral development with a tool for higher accuracy and sensitivity.

Biochemical changes in desmosomes of bovine muzzle epidermis during differentiation.

PubMed

Konohana, A; Konohana, I; Roberts, G P; Marks, R

1987-10-01

Biochemical changes taking place in desmosomes during differentiation have been studied. Bovine muzzle epidermis was sliced horizontally into 6 layers, 0.2 mm thick, and desmosomes were isolated from each layer. These were then analyzed by polyacrylamide gel electrophoresis. The electrophoretic patterns of desmosomal proteins from the 6 layers were found to be qualitatively similar to each other, but there was an increase in the ratio of the amount of 150 kD glycoprotein (desmoglein I) relative to 240 and 210 kD proteins (desmoplakins) in the upper layers of the epidermis. This finding was supported by the similar increase observed in electrophoretic patterns of proteins extracted directly from each layer of the epidermis in electrophoretic sample buffer. In order to study the fate of desmosomal components in the stratum corneum, serial skin surface biopsies were stained with antisera against desmosomal components using indirect immunofluorescence techniques. This experiment showed that desmosomal proteins and glycoproteins persist in the stratum corneum but quantitatively decrease in the outer layers. This decrease may play a significant role in desquamation.

Interface-induced multiferroism by design in complex oxide superlattices

PubMed Central

Guo, Hangwen; Wang, Zhen; Dong, Shuai; Ghosh, Saurabh; Saghayezhian, Mohammad; Chen, Lina; Weng, Yakui; Herklotz, Andreas; Ward, Thomas Z.; Jin, Rongying; Pantelides, Sokrates T.; Zhu, Yimei; Zhang, Jiandi; Plummer, E. W.

2017-01-01

Interfaces between materials present unique opportunities for the discovery of intriguing quantum phenomena. Here, we explore the possibility that, in the case of superlattices, if one of the layers is made ultrathin, unexpected properties can be induced between the two bracketing interfaces. We pursue this objective by combining advanced growth and characterization techniques with theoretical calculations. Using prototype La2/3Sr1/3MnO3 (LSMO)/BaTiO3 (BTO) superlattices, we observe a structural evolution in the LSMO layers as a function of thickness. Atomic-resolution EM and spectroscopy reveal an unusual polar structure phase in ultrathin LSMO at a critical thickness caused by interfacing with the adjacent BTO layers, which is confirmed by first principles calculations. Most important is the fact that this polar phase is accompanied by reemergent ferromagnetism, making this system a potential candidate for ultrathin ferroelectrics with ferromagnetic ordering. Monte Carlo simulations illustrate the important role of spin–lattice coupling in LSMO. These results open up a conceptually intriguing recipe for developing functional ultrathin materials via interface-induced spin–lattice coupling. PMID:28607082

Interface-induced multiferroism by design in complex oxide superlattices

DOE PAGES

Guo, Hangwen; Wang, Zhen; Dong, Shuai; ...

2017-05-19

Interfaces between materials present unique opportunities for the discovery of intriguing quantum phenomena. Here, we explore the possibility that, in the case of superlattices, if one of the layers is made ultrathin, unexpected properties can be induced between the two bracketing interfaces. We pursue this objective by combining advanced growth and characterization techniques with theoretical calculations. Using prototype La 2/3Sr 1/3MnO 3 (LSMO)/BaTiO 3 (BTO) superlattices, we observe a structural evolution in the LSMO layers as a function of thickness. Atomic-resolution EM and spectroscopy reveal an unusual polar structure phase in ultrathin LSMO at a critical thickness caused by interfacingmore » with the adjacent BTO layers, which is confirmed by first principles calculations. Most important is the fact that this polar phase is accompanied by reemergent ferromagnetism, making this system a potential candidate for ultrathin ferroelectrics with ferromagnetic ordering. Monte Carlo simulations illustrate the important role of spin–lattice coupling in LSMO. These results open up a conceptually intriguing recipe for developing functional ultrathin materials via interface-induced spin–lattice coupling.« less

Purification and characterization of tannase from Paecilomyces variotii: hydrolysis of tannic acid using immobilized tannase.

PubMed

Mahendran, B; Raman, N; Kim, D-J

2006-04-01

An extracellular tannase (tannin acyl hydrolase) was isolated from Paecilomyces variotii and purified from cell-free culture filtrate using ammonium sulfate precipitation followed by ion exchange and gel filtration chromatography. Fractional precipitation of the culture filtrate with ammonium sulfate yielded 78.7% with 13.6-folds purification, and diethylaminoethyl-cellulose column chromatography and gel filtration showed 19.4-folds and 30.5-folds purifications, respectively. Molecular mass of tannase was found 149.8 kDa through native polyacrylamide gel electrophoresis (PAGE) analysis. Sodium dodecyl sulphate-PAGE revealed that the purified tannase was a monomeric enzyme with a molecular mass of 45 kDa. Temperature of 30 to 50 degrees C and pH of 5.0 to 7.0 were optimum for tannase activity and stability. Tannase immobilized on alginate beads could hydrolyze tannic acid even after extensive reuse and retained about 85% of the initial activity. Thin layer chromatography, high performance liquid chromatography, and (1)H-nuclear magnetic resonance spectral analysis confirmed that gallic acid was formed as a byproduct during hydrolysis of tannic acid.

Simulating Electrophoresis.

ERIC Educational Resources Information Center

Moertel, Cheryl; Frutiger, Bruce

1996-01-01

Describes a DNA fingerprinting simulation that uses vegetable food coloring and plastic food containers instead of DNA and expensive gel electrophoresis chambers. Allows students to decipher unknown combinations of dyes in a method similar to that used to decipher samples of DNA in DNA fingerprint techniques. (JRH)

Development of a highly sensitive three-dimensional gel electrophoresis method for characterization of monoclonal protein heterogeneity.

PubMed

Nakano, Keiichi; Tamura, Shogo; Otuka, Kohei; Niizeki, Noriyasu; Shigemura, Masahiko; Shimizu, Chikara; Matsuno, Kazuhiko; Kobayashi, Seiichi; Moriyama, Takanori

2013-07-15

Three-dimensional gel electrophoresis (3-DE), which combines agarose gel electrophoresis and isoelectric focusing/SDS-PAGE, was developed to characterize monoclonal proteins (M-proteins). However, the original 3-DE method has not been optimized and its specificity has not been demonstrated. The main goal of this study was to optimize the 3-DE procedure and then compare it with 2-DE. We developed a highly sensitive 3-DE method in which M-proteins are extracted from a first-dimension agarose gel, by diffusing into 150 mM NaCl, and the recovery of M-proteins was 90.6%. To validate the utility of the highly sensitive 3-DE, we compared it with the original 3-DE method. We found that highly sensitive 3-DE provided for greater M-protein recovery and was more effective in terms of detecting spots on SDS-PAGE gels than the original 3-DE. Moreover, highly sensitive 3-DE separates residual normal IgG from M-proteins, which could not be done by 2-DE. Applying the highly sensitive 3-DE to clinical samples, we found that the characteristics of M-proteins vary tremendously between individuals. We believe that our highly sensitive 3-DE method described here will prove useful in further studies of the heterogeneity of M-proteins. Copyright © 2013 Elsevier Inc. All rights reserved.

High quality Ge epilayer on Si (1 0 0) with an ultrathin Si1-x Ge x /Si buffer layer by RPCVD

NASA Astrophysics Data System (ADS)

Chen, Da; Guo, Qinglei; Zhang, Nan; Xu, Anli; Wang, Bei; Li, Ya; Wang, Gang

2017-07-01

The authors report a method to grow high quality strain-relaxed Ge epilayer on a combination of low temperature Ge seed layer and Si1-x Ge x /Si superlattice buffer layer by reduced pressure chemical vapor deposition system without any subsequent annealing treatment. Prior to the growth of high quality Ge epilayer, an ultrathin Si1-x Ge x /Si superlattice buffer layer with the thickness of 50 nm and a 460 nm Ge seed layer were deposited successively at low temperature. Then an 840 nm Ge epilayer was grown at high deposition rate with the surface root-mean-square roughness of 0.707 nm and threading dislocation density of 2.5  ×  106 cm-2, respectively. Detailed investigations of the influence of ultrathin low-temperature Si1-x Ge x /Si superlattice buffer layer on the quality of Ge epilayer were performed, which indicates that the crystalline quality of Ge epilayer can be significantly improved by enhancing the Ge concentration of Si1-x Ge x /Si superlattice buffer layer.

Perpendicular magnetic tunnel junctions with Mn-modified ultrathin MnGa layer

NASA Astrophysics Data System (ADS)

Suzuki, K. Z.; Miura, Y.; Ranjbar, R.; Bainsla, L.; Ono, A.; Sasaki, Y.; Mizukami, S.

2018-02-01

Perpendicular magnetic tunnel junctions (p-MTJs) with a MgO barrier and a 1-nm-thick MnGa electrode were investigated by inserting several monolayers (MLs) of Mn. The tunnel magnetoresistance (TMR) ratio systematically increased when increasing the Mn layer thickness with a maximum of 18 (38.4)% at 300 (5) K for a Mn layer thickness of 0.6-0.8 nm. This ratio is five times higher compared to that without the Mn layer. The perpendicular magnetic anisotropy (PMA) field and the PMA constant of the ultrathin MnGa layer also increased up to 62-90 kOe and 6.2-11.3 Merg/cm3, respectively, with an increase in the Mn interlayer thickness, even for the ultrathin regime of the MnGa layer. For p-MTJs showing a high TMR and PMA, electron microscopy indicated the presence of 3-4 MLs of Mn at the MnGa/MgO interface; thus, the Mn modification enhanced the TMR as well as improved the PMA. This may be a promising finding to develop a Mn-based free layer for spin-transfer-torque devices for high-recording-density magnetoresistive random access memory and a sub-THz oscillator/detector.

Native and sodium dodecyl sulfate-capillary gel electrophoresis of proteins on a single microchip.

PubMed

Tsai, Shuo-Wen; Loughran, Michael; Suzuki, Hiroaki; Karube, Isao

2004-02-01

Simultaneous electrophoresis of both native and Sodium dodecyl sulfate (SDS) proteins was observed on a single microchip within 20 min. The capillary array prevented lateral diffusion of SDS components and avoided cross contamination of native protein samples. The planar sputtered electrode format provided a more uniform distribution of separation voltage into each of the 36 parallel microchannel capillaries than platinum wire electrodes commonly used in conventional electrophoresis. The customized geometry of the stacking capillary machined into the cover plate of the microchip facilitated reproducible sample injection without the requirement for stacking gel. Polyimide served as a mask and facilitated insulation of the anode and cathode to prevent electrode lift off and deterioration during continuous electrophoresis, even at a constant current of 8 mA. Improved protein separation was observed during capillary electrophoresis at lower currents. Ferguson plot analysis confirmed the electrophoretic mobility of native globular proteins in accordance with their charge and size. Corresponding Ferguson plot analysis of SDS-associated proteins on the same chip confirmed separation of marker proteins according to their molecular weight.

Decorative power generating panels creating angle insensitive transmissive colors

PubMed Central

Lee, Jae Yong; Lee, Kyu-Tae; Seo, Sungyong; Guo, L. Jay

2014-01-01

We present ultra-thin (6 to 31 nm) undoped amorphous silicon/organic hybrid solar cell structure, which can transmit desired color of light. The transmitted colors show great angular tolerance due to the negligible optical phase associated with light propagating in ultra-thin amorphous silicon (a-Si) layers. We achieved the power conversion efficiency of the hybrid cells up to 2 %; and demonstrated that most of the absorbed photons in the undoped a-Si layer contributed to the extracted electric charges due to the suppressed electron-hole recombination in the ultra-thin a-Si layer. We also show the resonance is invariant with respect to the angle of incidence up to ±70° regardless of the polarization of the incident light. Our exploration provides a design to realize energy harvesting colored photovoltaic panels for innovative applications. PMID:24577075

Decorative power generating panels creating angle insensitive transmissive colors

NASA Astrophysics Data System (ADS)

Lee, Jae Yong; Lee, Kyu-Tae; Seo, Sungyong; Guo, L. Jay

2014-02-01

We present ultra-thin (6 to 31 nm) undoped amorphous silicon/organic hybrid solar cell structure, which can transmit desired color of light. The transmitted colors show great angular tolerance due to the negligible optical phase associated with light propagating in ultra-thin amorphous silicon (a-Si) layers. We achieved the power conversion efficiency of the hybrid cells up to 2 %; and demonstrated that most of the absorbed photons in the undoped a-Si layer contributed to the extracted electric charges due to the suppressed electron-hole recombination in the ultra-thin a-Si layer. We also show the resonance is invariant with respect to the angle of incidence up to +/-70° regardless of the polarization of the incident light. Our exploration provides a design to realize energy harvesting colored photovoltaic panels for innovative applications.

Decorative power generating panels creating angle insensitive transmissive colors.

PubMed

Lee, Jae Yong; Lee, Kyu-Tae; Seo, Sungyong; Guo, L Jay

2014-02-28

We present ultra-thin (6 to 31 nm) undoped amorphous silicon/organic hybrid solar cell structure, which can transmit desired color of light. The transmitted colors show great angular tolerance due to the negligible optical phase associated with light propagating in ultra-thin amorphous silicon (a-Si) layers. We achieved the power conversion efficiency of the hybrid cells up to 2 %; and demonstrated that most of the absorbed photons in the undoped a-Si layer contributed to the extracted electric charges due to the suppressed electron-hole recombination in the ultra-thin a-Si layer. We also show the resonance is invariant with respect to the angle of incidence up to ± 70° regardless of the polarization of the incident light. Our exploration provides a design to realize energy harvesting colored photovoltaic panels for innovative applications.

Low-temperature atomic layer epitaxy of AlN ultrathin films by layer-by-layer, in-situ atomic layer annealing.

PubMed

Shih, Huan-Yu; Lee, Wei-Hao; Kao, Wei-Chung; Chuang, Yung-Chuan; Lin, Ray-Ming; Lin, Hsin-Chih; Shiojiri, Makoto; Chen, Miin-Jang

2017-01-03

Low-temperature epitaxial growth of AlN ultrathin films was realized by atomic layer deposition (ALD) together with the layer-by-layer, in-situ atomic layer annealing (ALA), instead of a high growth temperature which is needed in conventional epitaxial growth techniques. By applying the ALA with the Ar plasma treatment in each ALD cycle, the AlN thin film was converted dramatically from the amorphous phase to a single-crystalline epitaxial layer, at a low deposition temperature of 300 °C. The energy transferred from plasma not only provides the crystallization energy but also enhances the migration of adatoms and the removal of ligands, which significantly improve the crystallinity of the epitaxial layer. The X-ray diffraction reveals that the full width at half-maximum of the AlN (0002) rocking curve is only 144 arcsec in the AlN ultrathin epilayer with a thickness of only a few tens of nm. The high-resolution transmission electron microscopy also indicates the high-quality single-crystal hexagonal phase of the AlN epitaxial layer on the sapphire substrate. The result opens a window for further extension of the ALD applications from amorphous thin films to the high-quality low-temperature atomic layer epitaxy, which can be exploited in a variety of fields and applications in the near future.

Low-temperature atomic layer epitaxy of AlN ultrathin films by layer-by-layer, in-situ atomic layer annealing

PubMed Central

Shih, Huan-Yu; Lee, Wei-Hao; Kao, Wei-Chung; Chuang, Yung-Chuan; Lin, Ray-Ming; Lin, Hsin-Chih; Shiojiri, Makoto; Chen, Miin-Jang

2017-01-01

Low-temperature epitaxial growth of AlN ultrathin films was realized by atomic layer deposition (ALD) together with the layer-by-layer, in-situ atomic layer annealing (ALA), instead of a high growth temperature which is needed in conventional epitaxial growth techniques. By applying the ALA with the Ar plasma treatment in each ALD cycle, the AlN thin film was converted dramatically from the amorphous phase to a single-crystalline epitaxial layer, at a low deposition temperature of 300 °C. The energy transferred from plasma not only provides the crystallization energy but also enhances the migration of adatoms and the removal of ligands, which significantly improve the crystallinity of the epitaxial layer. The X-ray diffraction reveals that the full width at half-maximum of the AlN (0002) rocking curve is only 144 arcsec in the AlN ultrathin epilayer with a thickness of only a few tens of nm. The high-resolution transmission electron microscopy also indicates the high-quality single-crystal hexagonal phase of the AlN epitaxial layer on the sapphire substrate. The result opens a window for further extension of the ALD applications from amorphous thin films to the high-quality low-temperature atomic layer epitaxy, which can be exploited in a variety of fields and applications in the near future. PMID:28045075

Continuous monitoring of enzymatic activity within native electrophoresis gels: Application to mitochondrial oxidative phosphorylation complexes

PubMed Central

Covian, Raul; Chess, David; Balaban, Robert S.

2012-01-01

Native gel electrophoresis allows the separation of very small amounts of protein complexes while retaining aspects of their activity. In-gel enzymatic assays are usually performed by using reaction-dependent deposition of chromophores or light scattering precipitates quantified at fixed time points after gel removal and fixation, limiting the ability to analyze enzyme reaction kinetics. Herein, we describe a custom reaction chamber with reaction media recirculation and filtering and an imaging system that permits the continuous monitoring of in-gel enzymatic activity even in the presence of turbidity. Images were continuously collected using time-lapse high resolution digital imaging, and processing routines were developed to obtain kinetic traces of the in-gel activities and analyze reaction time courses. This system also permitted the evaluation of enzymatic activity topology within the protein bands of the gel. This approach was used to analyze the reaction kinetics of two mitochondrial complexes in native gels. Complex IV kinetics showed a short initial linear phase where catalytic rates could be calculated, whereas Complex V activity revealed a significant lag phase followed by two linear phases. The utility of monitoring the entire kinetic behavior of these reactions in native gels, as well as the general application of this approach, is discussed. PMID:22975200

Continuous monitoring of enzymatic activity within native electrophoresis gels: application to mitochondrial oxidative phosphorylation complexes.

PubMed

Covian, Raul; Chess, David; Balaban, Robert S

2012-12-01

Native gel electrophoresis allows the separation of very small amounts of protein complexes while retaining aspects of their activity. In-gel enzymatic assays are usually performed by using reaction-dependent deposition of chromophores or light-scattering precipitates quantified at fixed time points after gel removal and fixation, limiting the ability to analyze the enzyme reaction kinetics. Herein, we describe a custom reaction chamber with reaction medium recirculation and filtering and an imaging system that permits the continuous monitoring of in-gel enzymatic activity even in the presence of turbidity. Images were continuously collected using time-lapse high-resolution digital imaging, and processing routines were developed to obtain kinetic traces of the in-gel activities and analyze reaction time courses. This system also permitted the evaluation of enzymatic activity topology within the protein bands of the gel. This approach was used to analyze the reaction kinetics of two mitochondrial complexes in native gels. Complex IV kinetics showed a short initial linear phase in which catalytic rates could be calculated, whereas Complex V activity revealed a significant lag phase followed by two linear phases. The utility of monitoring the entire kinetic behavior of these reactions in native gels, as well as the general application of this approach, is discussed. Published by Elsevier Inc.

Coexistence of Topological Edge State and Superconductivity in Bismuth Ultrathin Film.

PubMed

Sun, Hao-Hua; Wang, Mei-Xiao; Zhu, Fengfeng; Wang, Guan-Yong; Ma, Hai-Yang; Xu, Zhu-An; Liao, Qing; Lu, Yunhao; Gao, Chun-Lei; Li, Yao-Yi; Liu, Canhua; Qian, Dong; Guan, Dandan; Jia, Jin-Feng

2017-05-10

Ultrathin freestanding bismuth film is theoretically predicted to be one kind of two-dimensional topological insulators. Experimentally, the topological nature of bismuth strongly depends on the situations of the Bi films. Film thickness and interaction with the substrate often change the topological properties of Bi films. Using angle-resolved photoemission spectroscopy, scanning tunneling microscopy or spectroscopy and first-principle calculation, the properties of Bi(111) ultrathin film grown on the NbSe 2 superconducting substrate have been studied. We find the band structures of the ultrathin film is quasi-freestanding, and one-dimensional edge state exists on Bi(111) film as thin as three bilayers. Superconductivity is also detected on different layers of the film and the pairing potential exhibits an exponential decay with the layer thicknesses. Thus, the topological edge state can coexist with superconductivity, which makes the system a promising platform for exploring Majorana Fermions.

Controllable synthesis of ultrathin vanadium oxide nanobelts via an EDTA-mediated hydrothermal process

NASA Astrophysics Data System (ADS)

Yu-Xiang, Qin; Cheng, Liu; Wei-Wei, Xie; Meng-Yang, Cui

2016-02-01

Ultrathin VO2 nanobelts with rough alignment features are prepared on the induction layer-coated substrates by an ethylenediaminetetraacetic acid (EDTA)-mediated hydrothermal process. EDTA acts as a chelating reagent and capping agent to facilitate the one-dimensional (1D) preferential growth of ultrathin VO2 nanobelts with high crystallinities and good uniformities. The annealed induction layer and concentration of EDTA are found to play crucial roles in the formation of aligned and ultrathin nanobelts. Variation in EDTA concentration can change the VO2 morphology of ultrathin nanobelts into that of thick nanoplates. Mild annealing of ultrathin VO2 nanobelts at 350 °C in air results in the formation of V2O5 nanobelts with a nearly unchanged ultrathin structure. The nucleation and growth mechanism involved in the formations of nanobelts and nanoplates are proposed. The ethanol gas sensing properties of the V2O5 nanobelt networks-based sensor are investigated in a temperature range from 100 °C to 300 °C over ethanol concentrations ranging from 3 ppm to 500 ppm. The results indicate that the V2O5 nanobelt network sensor exhibits high sensitivity, good reversibility, and fast response-recovery characteristics with an optimal working temperature of 250 °C. Project supported by the National Natural Science Foundation of China (Grant Nos. 61274074, 61271070, and 61574100).

The Make 2D-DB II package: conversion of federated two-dimensional gel electrophoresis databases into a relational format and interconnection of distributed databases.

PubMed

Mostaguir, Khaled; Hoogland, Christine; Binz, Pierre-Alain; Appel, Ron D

2003-08-01

The Make 2D-DB tool has been previously developed to help build federated two-dimensional gel electrophoresis (2-DE) databases on one's own web site. The purpose of our work is to extend the strength of the first package and to build a more efficient environment. Such an environment should be able to fulfill the different needs and requirements arising from both the growing use of 2-DE techniques and the increasing amount of distributed experimental data.

The Use of Gel Electrophoresis to Study the Reactions of Activated Amino Acids with Oligonucleotides

NASA Technical Reports Server (NTRS)

Zieboll, Gerhard; Orgel, Leslie E.

1994-01-01

We have used gel electrophoresis to study the primary covalent addition of amino acids to oligonu-cleotides or their analogs and the subsequent addition of further molecules of the amino acids to generate peptides covalently linked to the oligonucleotides. We have surveyed the reactions of a variety of amino acids with the phosphoramidates derived from oligonucleotide 5 inches phosphates and ethylenediamine. We find that arginine and amino acids can interact with oligonucleotidesl through stacking interactions react most efficiently. D- and L-amino acids give indistinguishable families of products.

Direct detection of rutin-degrading isozymes with polyacrylamide gel electrophoresis.

PubMed

Li, Yuping; Deng, Dandan; Zhang, Xuebin; Zhang, Haina; Wang, Cong; Chen, Peng

2013-12-15

Rutin-degrading enzymes (RDEs) specifically hydrolyze the glycosidic linkages of rutin, producing quercetin and rutinose. Here we report a reliable and sensitive polyacrylamide gel electrophoresis and staining method for the detection of RDE isozymes, which is based on the aqueous solubility difference between rutin and quercetin, as well as the ultraviolet absorbance of quercetin. With this novel method, we achieved a detection limit of 12 ng with 107 U of RDE activity, enabling us to detect at least five RDE isozymes in tartary buckwheat seeds. Copyright © 2013 Elsevier Inc. All rights reserved.

Karyotype analysis of anuran trypanosomes by pulsed-field gradient gel electrophoresis.

PubMed

Lun, Z R; Desser, S S

1995-12-01

The chromosomes of 12 species and isolates of anuran trypanosomes were investigated by pulsed-field gradient gel electrophoresis. Twelve to 16 chromosomes ranging from 0.45 to 2.2 megabase pairs were found in each of these trypanosomes. Minichromosomes were not observed in any of the examined isolates. Results indicate that different species of anuran trypanosomes display distinct karyotype patterns, and that isolates from the same region are similar. Our findings also reveal that most chromosome profiles of these trypanosomes are in accordance with isoenzyme and random amplified polymorphic DNA analysis data.

Two Distinct Clones of Methicillin-Resistant Staphylococcus aureus (MRSA) with the Same USA300 Pulsed-Field Gel Electrophoresis Profile: a Potential Pitfall for Identification of USA300 Community-Associated MRSA▿

PubMed Central

Larsen, Anders Rhod; Goering, Richard; Stegger, Marc; Lindsay, Jodi A.; Gould, Katherine A.; Hinds, Jason; Sørum, Marit; Westh, Henrik; Boye, Kit; Skov, Robert

2009-01-01

Analysis of methicillin-resistant Staphylococcus aureus (MRSA) characterized as USA300 by pulsed-field gel electrophoresis identified two distinct clones. One was similar to community-associated USA300 MRSA (ST8-IVa, t008, and Panton-Valentine leukocidin positive). The second (ST8-IVa, t024, and PVL negative) had different molecular characteristics and epidemiology, suggesting independent evolution. We recommend spa typing and/or PCR to discriminate between the two clones. PMID:19759225

Ultrathin TiO2 layer coated-CdS spheres core-shell nanocomposite with enhanced visible-light photoactivity.

PubMed

Chen, Zhang; Xu, Yi-Jun

2013-12-26

Development of various strategies for controllable fabrication of core-shell nanocomposites (CSNs) with highly active photocatalytic performance has been attracting ever-increasing research attention. In particular, control of the ultrathin layer TiO2 shell in constructing CSNs in an aqueous phase is a significant but technologically challenging issue. Here, this paper demonstrates the interface assembly synthesis of CdS nanospheres@TiO2 core-shell photocatalyst via the electrostatic interaction of negatively charged water-stable titania precursor with positively charged CdS nanospheres (CdS NSPs), followed by the formation of the ultrathin-layer TiO2 shell through a facile refluxing process in aqueous phase. The as-formed CdS NSPs@TiO2 core-shell nanohybrid exhibits a high visible-light-driven photoactivity for selective transformation and reduction of heavy metal ions. The ultrathin TiO2 layer coated on CdS NSPs results in excellent light transmission property, enhanced adsorption capacity, and improved transfer of charge carriers and lifespan of photoinduced electron-hole pairs, which would prominently contribute to the significant photoactivity enhancement. It is anticipated that this facile aqueous-phase synthesis strategy could be extended to design a variety of more efficient CSN photocatalysts with controllable morphology toward target applications in diverse photoredox processes.

Analysis of soybean tissue culture protein dynamics using difference gel electrophoresis.

PubMed

Miernyk, Ján A; Jett, Alissa A; Johnston, Mark L

2016-01-01

Excised hypocotyls from developing soybean (Glycine max (L.) merr. cv. Jack) were cultivated on agar-solidified medium until callus formed. The calli were then propagated in liquid medium until stable, relatively uniform, finely-divided suspension cultures were obtained. Cells were typically transferred to fresh medium at 7-day intervals. Cultures were harvested by filtration five days (early log phase) or eight days (late log phase) after transfer. In order to evaluate dynamic changes, both intracellular and extracellular proteins were analyzed by 2-dimensional difference gel electrophoresis. Selected spots were subjected to in-gel tryptic-digestion and the resultant peptides were analyzed by nLC-MS/MS. In follow-up studies gel-free shot-gun analyses led to identification of 367 intracellular proteins and 188 extracellular proteins. The significance of the described research is two-fold. First a gel-based proteomics method was applied to the study of the dynamics of the secretome (extracellular proteins). Second, results of a shot-gun non-gel based proteomic survey of both cellular and extracellular proteins are presented. Published by Elsevier B.V.

System for loading slab-gel holders for electrophoresis separation

DOEpatents

Anderson, Norman G.; Anderson, Norman L.

1979-01-01

A slab-gel loading system includes a prismatic chamber for filling a plurality of slab-gel holders simultaneously. Each slab-gel holder comprises a pair of spaced apart plates defining an intermediate volume for gel containment. The holders are vertically positioned in the chamber with their major surfaces parallel to the chamber end walls. A liquid inlet is provided at the corner between the bottom and a side wall of the chamber for distributing a polymerizable monomer solution or a coagulable colloidal solution into each of the holders. The chamber is rotatably supported so that filling can begin with the corner having the liquid inlet directed downwardly such that the solution is gently funneled upwardly, without mixing, along the diverging side and bottom surfaces. As filling proceeds, the chamber is gradually rotated to position the bottom wall in a horizontal mode. The liquid filling means includes a plastic envelope with a septum dividing it into two compartments for intermixing two solutions of different density and thereby providing a liquid flow having a density gradient. The resulting gels have a density gradient between opposite edges for subsequent use in electrophoresis separations.

Agarose electrophoresis of DNA in discontinuous buffers, using a horizontal slab apparatus and a buffer system with improved properties.

PubMed

Zsolnai, A; Orbán, L; Chrambach, A

1993-03-01

Using a horizontal slab apparatus with a buffer in the reservoirs at the level of the gel ("sea-level electrophoresis"), the retrograde discontinuous buffer system reported by Wiltfang et al. for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins was applied to DNA electrophoresis. This application yielded the advantages of an increased displacement rate of the moving boundary front and a decrease in the concentration of the counterion base in the resolving phase, which yielded reduced relative mobility values at equivalent gel concentrations and practicable low buffer concentrations. The change of relative mobilities (Rf) with a variation of field strength is decreased compared to that of the migration rate in the continuous Tris-boric-acid-EDTA (TBE) buffer and thus the robustness of the system is improved, as well as the efficiency of separation. The system of Wiltfang et al. has in common with previously described discontinuous DNA system, that it is able to stack DNA from dilute samples and is insensitive to sample components with lower net mobilities than DNA, such as acetate. However, the variance of Rf at constant current density in the discontinuous buffer system is not improved over that of the migration rate at constant field strength in the continuous TBE buffer.

Attempt to run urinary protein electrophoresis using capillary technique.

PubMed

Falcone, Michele

2014-10-01

The study of urinary protein has a predominant place in the diagnosis of kidney disease. The most common technique is agarose gel electrophoresis (AGE). For several years, the technique of choice applied to the analysis of serum proteins has been CE, a system that uses capillary fused silica, subjected to high voltage to separate and measure serum proteins. The purpose of this paper was to perform capillary electrophoresis on urinary proteins which, at present, are not interpretable due to the many nonspecific peaks visible when using gel electrophoresis. In order to carry out our research, we used a capillary V8 analyzer together with an agarose gel system from the same company. AGE was taken as the reference method, for which urine was used without any pretreatment. For the V8 system, urine was subjected to purification on granular-activated carbon and then inserted into the V8 analyzer, selecting a program suitable for liquids with low protein content. We examined 19 urine samples collected over 24 hrs from both hospitalized and external patients with different types of proteinuria plus a serum diluted 1/61 considered as a control to recognize the bands. Both methods showed the same protein fractions and classified the proteinuria in a similar way. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Corrosion resistance of monolayer hexagonal boron nitride on copper

PubMed Central

Mahvash, F.; Eissa, S.; Bordjiba, T.; Tavares, A. C.; Szkopek, T.; Siaj, M.

2017-01-01

Hexagonal boron nitride (hBN) is a layered material with high thermal and chemical stability ideal for ultrathin corrosion resistant coatings. Here, we report the corrosion resistance of Cu with hBN grown by chemical vapor deposition (CVD). Cyclic voltammetry measurements reveal that hBN layers inhibit Cu corrosion and oxygen reduction. We find that CVD grown hBN reduces the Cu corrosion rate by one order of magnitude compared to bare Cu, suggesting that this ultrathin layer can be employed as an atomically thin corrosion-inhibition coating. PMID:28191822

NOL specular spin-valve heads using an ultrathin CoFe free layer

NASA Astrophysics Data System (ADS)

Fukuzawa, Hideaki; Koi, Katsuhiko; Tomita, Hiroshi; Fuke, Hiromi Niu; Kamiguchi, Yuzo; Iwasaki, Hitoshi; Sahashi, Masashi

2001-10-01

This paper reports the film and head performance of specular spin valves with nano-oxide layer (NOL-SPSV). A large MR ratio of 17% was obtained by using an ultrathin CoFe free layer with a high conductance Cu layer, which decreases the sense current field of a free layer and brings good soft magnetic characteristics. Prototype heads with a read track width of 0.47-0.61 μm were fabricated by using NOL-SPSV films with an MR ratio of 14-15%, Hua˜400 Oe, and Hc˜5 Oe. High output signal voltage of 8-11 mV/μm was realized in the NOL-SPSV heads.

Suppression of surface segregation of the phosphorous δ-doping layer by insertion of an ultra-thin silicon layer for ultra-shallow Ohmic contacts on n-type germanium

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

Yamada, Michihiro; Uematsu, Masashi; Itoh, Kohei M., E-mail: kitoh@appi.keio.ac.jp

2015-09-28

We demonstrate the formation of abrupt phosphorus (P) δ-doping profiles in germanium (Ge) by the insertion of ultra-thin silicon (Si) layers. The Si layers at the δ-doping region significantly suppress the surface segregation of P during the molecular beam epitaxial growth of Ge and high-concentration active P donors are confined within a few nm of the initial doping position. The current-voltage characteristics of the P δ-doped layers with Si insertion show excellent Ohmic behaviors with low enough resistivity for ultra-shallow Ohmic contacts on n-type Ge.

"Self-Peel-Off" Transfer Produces Ultrathin Polyvinylidene-Fluoride-Based Flexible Nanodevices.

PubMed

Tai, Yanlong; Lubineau, Gilles

2017-04-01

Here, a new strategy, self-peel-off transfer, for the preparation of ultrathin flexible nanodevices made from polyvinylidene-fluoride (PVDF) is reported. In this process, a functional pattern of nanoparticles is transferred via peeling from a temporary substrate to the final PVDF film. This peeling process takes advantage of the differences in the work of adhesion between the various layers (the PVDF layer, the nanoparticle-pattern layer and the substrate layer) and of the high stresses generated by the differential thermal expansion of the layers. The work of adhesion is mainly guided by the basic physical/chemical properties of these layers and is highly sensitive to variations in temperature and moisture in the environment. The peeling technique is tested on a variety of PVDF-based functional films using gold/palladium nanoparticles, carbon nanotubes, graphene oxide, and lithium iron phosphate. Several PVDF-based flexible nanodevices are prepared, including a single-sided wireless flexible humidity sensor in which PVDF is used as the substrate and a double-sided flexible capacitor in which PVDF is used as the ferroelectric layer and the carrier layer. Results show that the nanodevices perform with high repeatability and stability. Self-peel-off transfer is a viable preparation strategy for the design and fabrication of flexible, ultrathin, and light-weight nanodevices.

[Study on diversity of protein between Houttuynia cordata plant].

PubMed

Zhang, Xi-li; He, Fu-yuan; Wang, Hai-qin; Yang, Yan-tao; Shi, Ji-lian; Liu, Wen-long; Li, Shun-xiang

2013-12-01

To reveal protein diversity between the same batch of fresh Houttuynia cordata in the same GAP base,and to lay the foundation construction for "node metabolic network". Three methods including the Ramagli improved Bradford law, SDS-PAGE gel electrophoresis method and double wavelength thin-layer scanning method were used to study the total protein content diversity, protein species diversity and various kinds of content variability. The molecular weight of 53 plant protein mostly concentrated in the range of 6.5-97.2 kDa, the species diversity was not obvious with main performance for banding color shades; The RSD of zero moment (AUCT), first moment (MCRTT) and second moment (VCRTT) in protein electrophoresis banding was 40.92%, 6.01% and 18.57%, respectively. There is rich diversity in different Houttuynia cordata plant in the same GAP base, which provides basis for the foundation of subsequent key protease search, "node metabolic network" construction, and study on the Chinese medicine quality stability.

A DFT study on NEA GaN photocathode with an ultrathin n-type Si-doped GaN cap layer

NASA Astrophysics Data System (ADS)

Xia, Sihao; Liu, Lei; Kong, Yike; Diao, Yu

2016-10-01

Due to the drawbacks of conventional negative electron affinity (NEA) GaN photocathodes activated by Cs or Cs/O, a new-type NEA GaN photocathodes with heterojunction surface dispense with Cs activation are proposed. This structure can be obtained through the coverage of an ultrathin n-type Si-doped GaN cap layer on the p-type Mg-doped GaN emission layer. The influences of the cap layer on the photocathode are calculated using DFT. This study indicates that the n-type cap layer can promote the photoemission characteristics of GaN photocathode and demonstrates the probability of the preparation of a NEA GaN photocathode with an n-type cap layer.

The Ultrathin Limit and Dead-layer Effects in Local Polarization Switching of BiFeO3

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

Maksymovych, Petro; Huijben, Mark; Pan, Minghu

Using piezoresponse force microscopy in ultra-high vacuum, polarization switching has been detected and quantified in epitaxial BiFeO3 films from 200 down to ~ 4 unit cells. Local remnant piezoresponse was used to infer the applied electric field inside the ferroelectric volume, and account for the elusive effect of dead-layers in ultrathin films. The dead-layer manifested itself in the slower than anticipated decrease of the switching bias with film thickness, yielding apparent Kay-Dunn scaling of the switching field, while the statistical analysis of hysteresis loops revealed lateral variation of the dead-layer with sub-10 nm resolution.

Precise, Self-Limited Epitaxy of Ultrathin Organic Semiconductors and Heterojunctions Tailored by van der Waals Interactions.

PubMed

Wu, Bing; Zhao, Yinghe; Nan, Haiyan; Yang, Ziyi; Zhang, Yuhan; Zhao, Huijuan; He, Daowei; Jiang, Zonglin; Liu, Xiaolong; Li, Yun; Shi, Yi; Ni, Zhenhua; Wang, Jinlan; Xu, Jian-Bin; Wang, Xinran

2016-06-08

Precise assembly of semiconductor heterojunctions is the key to realize many optoelectronic devices. By exploiting the strong and tunable van der Waals (vdW) forces between graphene and organic small molecules, we demonstrate layer-by-layer epitaxy of ultrathin organic semiconductors and heterostructures with unprecedented precision with well-defined number of layers and self-limited characteristics. We further demonstrate organic p-n heterojunctions with molecularly flat interface, which exhibit excellent rectifying behavior and photovoltaic responses. The self-limited organic molecular beam epitaxy (SLOMBE) is generically applicable for many layered small-molecule semiconductors and may lead to advanced organic optoelectronic devices beyond bulk heterojunctions.

The laboratory technology of discrete molecular separation: the historical development of gel electrophoresis and the material epistemology of biomolecular science, 1945-1970.

PubMed

Chiang, Howard Hsueh-hao

2009-01-01

Preparative and analytical methods developed by separation scientists have played an important role in the history of molecular biology. One such early method is gel electrophoresis, a technique that uses various types of gel as its supporting medium to separate charged molecules based on size and other properties. Historians of science, however, have only recently begun to pay closer attention to this material epistemological dimension of biomolecular science. This paper substantiates the historiographical thread that explores the relationship between modern laboratory practice and the production of scientific knowledge. It traces the historical development of gel electrophoresis from the mid-1940s to the mid-1960s, with careful attention to the interplay between technical developments and disciplinary shifts, especially the rise of molecular biology in this time-frame. Claiming that the early 1950s marked a decisive shift in the evolution of electrophoretic methods from moving boundary to zone electrophoresis, I reconstruct various trajectories in which scientists such as Oliver Smithies sought out the most desirable solid supporting medium for electrophoretic instrumentation. Biomolecular knowledge, I argue, emerged in part from this process of seeking the most appropriate supporting medium that allowed for discrete molecular separation and visualization. The early 1950s, therefore, marked not only an important turning point in the history of separation science, but also a transformative moment in the history of the life sciences as the growth of molecular biology depended in part on the epistemological access to the molecular realm available through these evolving technologies.

A study on the optics of copper indium gallium (di)selenide (CIGS) solar cells with ultra-thin absorber layers.

PubMed

Xu, Man; Wachters, Arthur J H; van Deelen, Joop; Mourad, Maurice C D; Buskens, Pascal J P

2014-03-10

We present a systematic study of the effect of variation of the zinc oxide (ZnO) and copper indium gallium (di)selenide (CIGS) layer thickness on the absorption characteristics of CIGS solar cells using a simulation program based on finite element method (FEM). We show that the absorption in the CIGS layer does not decrease monotonically with its layer thickness due to interference effects. Ergo, high precision is required in the CIGS production process, especially when using ultra-thin absorber layers, to accurately realize the required thickness of the ZnO, cadmium sulfide (CdS) and CIGS layer. We show that patterning the ZnO window layer can strongly suppress these interference effects allowing a higher tolerance in the production process.

System and Method for Fabricating Super Conducting Circuitry on Both Sides of an Ultra-Thin Layer

NASA Technical Reports Server (NTRS)

Brown, Ari D. (Inventor); Mikula, Vilem (Inventor)

2017-01-01

A method of fabricating circuitry in a wafer includes depositing a superconducting metal on a silicon on insulator wafer having a handle wafer, coating the wafer with a sacrificial layer and bonding the wafer to a thermally oxide silicon wafer with a first epoxy. The method includes flipping the wafer, thinning the flipped wafer by removing a handle wafer, etching a buried oxide layer, depositing a superconducting layer, bonding the wafer to a thermally oxidized silicon wafer having a handle wafer using an epoxy, flipping the wafer again, thinning the flipped wafer, etching a buried oxide layer from the wafer and etching the sacrificial layer from the wafer. The result is a wafer having superconductive circuitry on both sides of an ultra-thin silicon layer.

Gel Electrophoresis of Gold-DNA Nanoconjugates

DOE PAGES

Pellegrino, T.; Sperling, R. A.; Alivisatos, A. P.; ...

2007-01-01

Gold-DNA conjugates were investigated in detail by a comprehensive gel electrophoresis study based on 1200 gels. A controlled number of single-stranded DNA of different length was attached specifically via thiol-Au bonds to phosphine-stabilized colloidal gold nanoparticles. Alternatively, the surface of the gold particles was saturated with single stranded DNA of different length either specifically via thiol-Au bonds or by nonspecific adsorption. From the experimentally determined electrophoretic mobilities, estimates for the effective diameters of the gold-DNA conjugates were derived by applying two different data treatment approaches. The first method is based on making a calibration curve for the relation between effectivemore » diameters and mobilities with gold nanoparticles of known diameter. The second method is based on Ferguson analysis which uses gold nanoparticles of known diameter as reference database. Our study shows that effective diameters derived from gel electrophoresis measurements are affected with a high error bar as the determined values strongly depend on the method of evaluation, though relative changes in size upon binding of molecules can be detected with high precision. Furthermore, in this study, the specific attachment of DNA via gold-thiol bonds to Au nanoparticles is compared to nonspecific adsorption of DNA. Also, the maximum number of DNA molecules that can be bound per particle was determined.« less

Fabrication of silicon-on-diamond substrate with an ultrathin SiO2 bonding layer

NASA Astrophysics Data System (ADS)

Nagata, Masahiro; Shirahama, Ryouya; Duangchan, Sethavut; Baba, Akiyoshi

2018-06-01

We proposed and demonstrated a sputter etching method to prepare both a flat surface (root-mean-square surface roughness of approximately 0.2–0.3 nm) and an ultrathin SiO2 bonding layer at an accuracy of approximately 5 nm in thickness to fabricate a silicon-on-diamond substrate (SOD). We also investigated a plasma activation method on a SiO2 surface using various gases. We found that O2 plasma activation is more suitable for the bonding between SiO2 and Si than N2 or Ar plasma activation. We speculate that the concentration of hydroxyl groups on the SiO2 surface was increased by O2 plasma activation. We fabricated the SOD substrate with an ultrathin (15 nm in thickness) SiO2 bonding layer using the sputter etching and O2 plasma activation methods.

Ultra-thin silicon oxide layers on crystalline silicon wafers: Comparison of advanced oxidation techniques with respect to chemically abrupt SiO2/Si interfaces with low defect densities

NASA Astrophysics Data System (ADS)

Stegemann, Bert; Gad, Karim M.; Balamou, Patrice; Sixtensson, Daniel; Vössing, Daniel; Kasemann, Martin; Angermann, Heike

2017-02-01

Six advanced oxidation techniques were analyzed, evaluated and compared with respect to the preparation of high-quality ultra-thin oxide layers on crystalline silicon. The resulting electronic and chemical SiO2/Si interface properties were determined by a combined x-ray photoemission (XPS) and surface photovoltage (SPV) investigation. Depending on the oxidation technique, chemically abrupt SiO2/Si interfaces with low densities of interface states were fabricated on c-Si either at low temperatures, at short times, or in wet-chemical environment, resulting in each case in excellent interface passivation. Moreover, the beneficial effect of a subsequent forming gas annealing (FGA) step for the passivation of the SiO2/Si interface of ultra-thin oxide layers has been proven. Chemically abrupt SiO2/Si interfaces have been shown to generate less interface defect states.

Morphology, mechanical stability, and protective properties of ultrathin gallium oxide coatings.

PubMed

Lawrenz, Frank; Lange, Philipp; Severin, Nikolai; Rabe, Jürgen P; Helm, Christiane A; Block, Stephan

2015-06-02

Ultrathin gallium oxide layers with a thickness of 2.8 ± 0.2 nm were transferred from the surface of liquid gallium onto solid substrates, including conjugated polymer poly(3-hexylthiophene) (P3HT). The gallium oxide exhibits high mechanical stability, withstanding normal pressures of up to 1 GPa in contact mode scanning force microscopy imaging. Moreover, it lowers the rate of photodegradation of P3HT by 4 orders of magnitude, as compared to uncovered P3HT. This allows us to estimate the upper limits for oxygen and water vapor transmission rates of 0.08 cm(3) m(-2) day(-1) and 0.06 mg m(-2) day(-1), respectively. Hence, similar to other highly functional coatings such as graphene, ultrathin gallium oxide layers can be regarded as promising candidates for protective layers in flexible organic (opto-)electronics and photovoltaics because they offer permeation barrier functionalities in conjunction with high optical transparency.

Spatially and momentum resolved energy electron loss spectra from an ultra-thin PrNiO{sub 3} layer

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

Kinyanjui, M. K., E-mail: michael.kinyanjui@uni-ulm.de; Kaiser, U.; Benner, G.

2015-05-18

We present an experimental approach which allows for the acquisition of spectra from ultra-thin films at high spatial, momentum, and energy resolutions. Spatially and momentum (q) resolved electron energy loss spectra have been obtained from a 12 nm ultra-thin PrNiO{sub 3} layer using a nano-beam electron diffraction based approach which enabled the acquisition of momentum resolved spectra from individual, differently oriented nano-domains and at different positions of the PrNiO{sub 3} thin layer. The spatial and wavelength dependence of the spectral excitations are obtained and characterized after the analysis of the experimental spectra using calculated dielectric and energy loss functions. The presentedmore » approach makes a contribution towards obtaining momentum-resolved spectra from nanostructures, thin film, heterostructures, surfaces, and interfaces.« less

Assessment of the microbial community in a constructed wetland that receives acid coal mine drainage

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

Nicomrat, D.; Dick, W.A.; Tuovinen, O.H.

2006-01-15

Constructed wetlands are used to treat acid drainage from surface or underground coal mines. However, little is known about the microbial communities in the receiving wetland cells. The purpose of this work was to characterize the microbial population present in a wetland that was receiving acid coal mine drainage (AMD). Samples were collected from the oxic sediment zone of a constructed wetland cell in southeastern Ohio that was treating acid drainage from an underground coal mine seep. Samples comprised Fe(Ill) precipitates and were pretreated with ammonium oxalate to remove interfering iron, and the DNA was extracted and purified by agarosemore » gel electrophoresis prior to amplification of portions of the 16S rRNA gene. Amplified products were separated by denaturing gradient gel electrophoresis and DNA from seven distinct bands was excised from the gel and sequenced. The sequences were matched to sequences in the GenBank bacterial 16S rDNA database. The DNA in two of the bands yielded matches with Acidithiobacillus ferrooxidans and the DNA in each of the remaining five bands was consistent with one of the following microorganisms: Acidithiobacillus thiooxidans, strain TRA3-20 (a eubacterium), strain BEN-4 (an arsenite-oxidizing bacterium), an Alcaligenes sp., and a Bordetella sp. Low bacterial diversity in these samples reflects the highly inorganic nature of the oxic sediment layer where high abundance of iron- and sulfur-oxidizing bacteria would be expected. The results we obtained by molecular methods supported our findings, obtained using culture methods, that the dominant microbial species in an acid receiving, oxic wetland are A. thiooxidans and A. ferrooxidans.« less

Single-Cell Western Blotting after Whole-Cell Imaging to Assess Cancer Chemotherapeutic Response

PubMed Central

2015-01-01

Intratumor heterogeneity remains a major obstacle to effective cancer therapy and personalized medicine. Current understanding points to differential therapeutic response among subpopulations of tumor cells as a key challenge to successful treatment. To advance our understanding of how this heterogeneity is reflected in cell-to-cell variations in chemosensitivity and expression of drug-resistance proteins, we optimize and apply a new targeted proteomics modality, single-cell western blotting (scWestern), to a human glioblastoma cell line. To acquire both phenotypic and proteomic data on the same, single glioblastoma cells, we integrate high-content imaging prior to the scWestern assays. The scWestern technique supports thousands of concurrent single-cell western blots, with each assay comprised of chemical lysis of single cells seated in microwells, protein electrophoresis from those microwells into a supporting polyacrylamide (PA) gel layer, and in-gel antibody probing. We systematically optimize chemical lysis and subsequent polyacrylamide gel electrophoresis (PAGE) of the single-cell lysate. The scWestern slides are stored for months then reprobed, thus allowing archiving and later analysis as relevant to sparingly limited, longitudinal cell specimens. Imaging and scWestern analysis of single glioblastoma cells dosed with the chemotherapeutic daunomycin showed both apoptotic (cleaved caspase 8- and annexin V-positive) and living cells. Intriguingly, living glioblastoma subpopulations show up-regulation of a multidrug resistant protein, P-glycoprotein (P-gp), suggesting an active drug efflux pump as a potential mechanism of drug resistance. Accordingly, linking of phenotype with targeted protein analysis with single-cell resolution may advance our understanding of drug response in inherently heterogeneous cell populations, such as those anticipated in tumors. PMID:25226230

Role of ultrathin metal fluoride layer in organic photovoltaic cells: mechanism of efficiency and lifetime enhancement.

PubMed

Lim, Kyung-Geun; Choi, Mi-Ri; Kim, Ji-Hoon; Kim, Dong Hun; Jung, Gwan Ho; Park, Yongsup; Lee, Jong-Lam; Lee, Tae-Woo

2014-04-01

Although rapid progress has been made recently in bulk heterojunction organic solar cells, systematic studies on an ultrathin interfacial layer at the electron extraction contact have not been conducted in detail, which is important to improve both the device efficiency and the lifetime. We find that an ultrathin BaF2 layer at the electron extraction contact strongly influences the open-circuit voltage (Voc ) as the nanomorphology evolves with increasing BaF2 thickness. A vacuum-deposited ultrathin BaF2 layer grows by island growth, so BaF2 layers with a nominal thickness less than that of single-coverage layer (≈3 nm) partially cover the polymeric photoactive layer. As the nominal thickness of the BaF2 layer increased to that of a single-coverage layer, the Voc and power conversion efficiency (PCE) of the organic photovoltaic cells (OPVs) increased but the short-circuit current remained almost constant. The fill factor and the PCE decreased abruptly as the thickness of the BaF2 layer exceeded that of a single-coverage layer, which was ascribed to the insulating nature of BaF2 . We find the major cause of the increased Voc observed in these devices is the lowered work function of the cathode caused by the reaction and release of Ba from thin BaF2 films upon deposition of Al. The OPV device with the BaF2 layer showed a slightly improved maximum PCE (4.0 %) and a greatly (approximately nine times) increased device half-life under continuous simulated solar irradiation at 100 mW cm(-2) as compared with the OPV without an interfacial layer (PCE=2.1 %). We found that the photodegradation of the photoactive layer was not a major cause of the OPV degradation. The hugely improved lifetime with cathode interface modification suggests a significant role of the cathode interfacial layer that can help to prolong device lifetimes. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The new horizon in 2D electrophoresis: new technology to increase resolution and sensitivity.

PubMed

Moche, Martin; Albrecht, Dirk; Maaß, Sandra; Hecker, Michael; Westermeier, Reiner; Büttner, Knut

2013-06-01

A principally new type of an electrophoresis setup for the second dimension of 2DE named HPE (high performance electrophoresis) has recently become available that provides excellent reproducibility much superior to traditional 2DE. It takes up ideas from early beginnings of 2DE which could not be satisfactory realized at that time. The new HPE system is in contrast to all other established systems a horizontal electrophoresis that employs a new type of precast polyacrylamide gels on film-backing and runs on a multilevel flatbed electrophoresis apparatus. In a systematic approach we compared its features to traditional 2DE for the cytosolic proteome of Bacillus subtilis. Not only the reproducibility is enhanced, but also nearly all qualitative parameters as resolution, sensitivity, the number of protein spots (25% more), and the number of different proteins (also additional 25%) are markedly increased. More than 200 proteins were exclusively found in HPE. This new electrophoresis system does not use buffer tanks. No glass plates are needed. Therefore handling of gels is greatly facilitated and very simple to use even for personnel with low technical skills. The new HPE system is technically at the beginnings and further development with increased performance can be expected. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Identification of column edges of DNA fragments by using K-means clustering and mean algorithm on lane histograms of DNA agarose gel electrophoresis images

NASA Astrophysics Data System (ADS)

Turan, Muhammed K.; Sehirli, Eftal; Elen, Abdullah; Karas, Ismail R.

2015-07-01

Gel electrophoresis (GE) is one of the most used method to separate DNA, RNA, protein molecules according to size, weight and quantity parameters in many areas such as genetics, molecular biology, biochemistry, microbiology. The main way to separate each molecule is to find borders of each molecule fragment. This paper presents a software application that show columns edges of DNA fragments in 3 steps. In the first step the application obtains lane histograms of agarose gel electrophoresis images by doing projection based on x-axis. In the second step, it utilizes k-means clustering algorithm to classify point values of lane histogram such as left side values, right side values and undesired values. In the third step, column edges of DNA fragments is shown by using mean algorithm and mathematical processes to separate DNA fragments from the background in a fully automated way. In addition to this, the application presents locations of DNA fragments and how many DNA fragments exist on images captured by a scientific camera.

A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis

PubMed Central

Huang, Kunlun; Zhang, Nan; Yuan, Yanfang; Shang, Ying; Luo, Yunbo

2012-01-01

In this study, a novel universal primer-multiplex-PCR (UP-M-PCR) method adding a universal primer (UP) in the multiplex PCR reaction system was described. A universal adapter was designed in the 5′-end of each specific primer pairs which matched with the specific DNA sequences for each template and also used as the universal primer (UP). PCR products were analyzed on sequencing gel electrophoresis (SGE) which had the advantage of exhibiting extraordinary resolution. This method overcame the disadvantages rooted deeply in conventional multiplex PCR such as complex manipulation, lower sensitivity, self-inhibition and amplification disparity resulting from different primers, and it got a high specificity and had a low detection limit of 0.1 ng for single kind of crops when screening the presence of genetically modified (GM) crops in mixture samples. The novel developed multiplex PCR assay with sequencing gel electrophoresis analysis will be useful in many fields, such as verifying the GM status of a sample irrespective of the crop and GM trait and so on. PMID:22272223

Automated one-step DNA sequencing based on nanoliter reaction volumes and capillary electrophoresis.

PubMed

Pang, H M; Yeung, E S

2000-08-01

An integrated system with a nano-reactor for cycle-sequencing reaction coupled to on-line purification and capillary gel electrophoresis has been demonstrated. Fifty nanoliters of reagent solution, which includes dye-labeled terminators, polymerase, BSA and template, was aspirated and mixed with the template inside the nano-reactor followed by cycle-sequencing reaction. The reaction products were then purified by a size-exclusion chromatographic column operated at 50 degrees C followed by room temperature on-line injection of the DNA fragments into a capillary for gel electrophoresis. Over 450 bases of DNA can be separated and identified. As little as 25 nl reagent solution can be used for the cycle-sequencing reaction with a slightly shorter read length. Significant savings on reagent cost is achieved because the remaining stock solution can be reused without contamination. The steps of cycle sequencing, on-line purification, injection, DNA separation, capillary regeneration, gel-filling and fluidic manipulation were performed with complete automation. This system can be readily multiplexed for high-throughput DNA sequencing or PCR analysis directly from templates or even biological materials.

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

PubMed Central

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

2012-01-01

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

Techniques for characterization and eradication of potato cyst nematode: a review.

PubMed

Bairwa, Aarti; Venkatasalam, E P; Sudha, R; Umamaheswari, R; Singh, B P

2017-09-01

Correct identification of species and pathotypes is must for eradication of potato cyst nematodes (PCN). The identification of PCN species after completing the life cycle is very difficult because it is based on morphological and morphometrical characteristics. Genetically different populations of PCN are morphologically same and differentiated based on the host differential study. Later on these traditional techniques have been replaced by biochemical techniques viz, one and two dimensional gel electrophoresis, capillary gel electrophoresis, isozymes, dot blot hybridization and isoelectric focusing etc. to distinguish both the species. One and two dimensional gel electrophoresis has used to examine inter- and intra-specific differences in proteins of Globodera rostochiensis and G. pallida . Now application of PCR and DNA based characterization techniques like RAPD, AFLP and RFLP are the important tools for differentiating inter- and intra specific variation in PCN and has given opportunities to accurate identification of PCN. For managing the PCN, till now we are following integrated pest management (IPM) strategies, however these strategies are not effective to eradicate the PCN. Therefore to eradicate the PCN we need noval management practices like RNAi (RNA interference) or Gene silencing.

Analysis of the structure of poly-3-hydroxybutyrate ultrathin fibers modified with iron (III) complex with tetraphenylporphyrin

NASA Astrophysics Data System (ADS)

Olkhov, A. A.; Karpova, S. G.; Lobanov, A. V.; Tyubaeva, P. M.; Artemov, N. S.; Iordansky, A. L.

2017-12-01

In the treatment of many infectious diseases and cancer, transdermal systems based on solid polymer matrices or gels containing functional substances with antiseptic (antibacterial) properties are often used. One of the most promising types of matrices with antiseptic properties are the ones of nano- and microfiber-bonded cloth obtained by electrospinning based on biopolymer poly(3-hydroxybutyrate). The present work investigates the effects of iron (III) complex with tetraphenylporphyrin and the influence on the geometry, crystalline order and molecular dynamics in the intercrystalline (amorphous phase) of ultrathin PHB fibers.

An effective placental cotyledons proteins extraction method for 2D gel electrophoresis.

PubMed

Tan, Niu J; Daim, Leona D J; Jamil, Amilia A M; Mohtarrudin, Norhafizah; Thilakavathy, Karuppiah

2017-03-01

Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Behavior of variable V3 region from 16S rDNA of lactic acid bacteria in denaturing gradient gel electrophoresis.

PubMed

Ercolini, D; Moschetti, G; Blaiotta, G; Coppola, S

2001-03-01

Separation of amplified V3 region from 16S rDNA by denaturing gradient gel electrophoresis (DGGE) was tested as a tool for differentiation of lactic acid bacteria commonly isolated from food. Variable V3 regions of 21 reference strains and 34 wild strains referred to species belonging to the genera Pediococcus, Enterococcus, Lactococcus, Lactobacillus, Leuconostoc, Weissella, and Streptococcus were analyzed. DGGE profiles obtained were species-specific for most of the cultures tested. Moreover, it was possible to group the remaining LAB reference strains according to the migration of their 16S V3 region in the denaturing gel. The results are discussed with reference to their potential in the analysis of LAB communities in food, besides shedding light on taxonomic aspects.

Purification and properties of trimethylamine oxide reductase from Salmonella typhimurium.

PubMed Central

Kwan, H S; Barrett, E L

1983-01-01

The major inducible trimethylamine oxide reductase was purified from Salmonella typhimurium LT2. The molecular weights of the native enzyme were estimated to be 332,000 by gel filtration and 170,000 by nondenaturing disc gel electrophoresis. In sodium dodecyl sulfate-gel electrophoresis, the enzyme formed a single band of molecular weight 84,000. The isoelectric point was 4.28. Maximum activity was at pH 5.65 and 45 degrees C. Reduced flavin mononucleotide, but not reduced flavin adenine dinucleotide, served as an electron donor. The Km for trimethylamine oxide was 0.89 mM and Vmax was 1,450 U/mg of protein. The enzyme reduced chlorate with a Km of 2.2 mM and a Vmax of 350 U/mg of protein. Images PMID:6350272

A noninvasive, direct real-time PCR method for sex determination in multiple avian species

USGS Publications Warehouse

Brubaker, Jessica L.; Karouna-Renier, Natalie K.; Chen, Yu; Jenko, Kathryn; Sprague, Daniel T.; Henry, Paula F.P.

2011-01-01

Polymerase chain reaction (PCR)-based methods to determine the sex of birds are well established and have seen few modifications since they were first introduced in the 1990s. Although these methods allowed for sex determination in species that were previously difficult to analyse, they were not conducive to high-throughput analysis because of the laboriousness of DNA extraction and gel electrophoresis. We developed a high-throughput real-time PCR-based method for analysis of sex in birds, which uses noninvasive sample collection and avoids DNA extraction and gel electrophoresis.

Isolation and purification of wheat germ agglutinin and analysis of its properties

NASA Astrophysics Data System (ADS)

Wang, Han

2017-12-01

In this paper, the wheat germ agglutinin was isolated and purified by affinity chromatography of chicken ovomucoid as ligand. The physicochemical properties were analyzed. The chicken ovomucoid was isolated from egg white and conjugated to affinity chromatography column agarose gel to prepare affinity adsorbent. The crude extract of wheat germ was freezedried by affinity chromatography. The physicochemical properties were analyzed by SDSpolyacrylamide gel electrophoresis and isoelectric focusing electrophoresis. And the relative molecular mass and isoelectric point of wheat germ agglutinin were obtained, and the high efficiency of purification of wheat germ agglutinin was proved by affinity chromatography.

Transient spectra study on photo-dynamics of curcumin

NASA Astrophysics Data System (ADS)

Qian, Tingting; Wang, Mei; Wang, Jiao; Zhu, Rongrong; He, Xiaolie; Sun, Xiaoyu; Sun, Dongmei; Wang, Qingxiu; Wang, ShiLong

2016-09-01

A novel mechanism of DNA damage induced by photosensitized curcumin (Cur) was explored using laser flash photolysis, pulse radiolysis and gel electrophoresis. Cur neutral radical (Currad) was confirmed as an identical product in photo-sensitization of Cur by laser flash photolysis and pulse radiolysis. A series of reaction rate constants between Currad and nucleic acid bases/nucleotides were determined by pulse radiolysis. Gel electrophoresis was carried out to investigate damage induced by photosensitized Cur to biologically active DNA. The results indicate that the damage to DNA may be caused by Currad produced from the photosensitization of Cur.

A subtle calculation method for nanoparticle’s molar extinction coefficient: The gift from discrete protein-nanoparticle system on agarose gel electrophoresis

NASA Astrophysics Data System (ADS)

Zhong, Ruibo; Yuan, Ming; Gao, Haiyang; Bai, Zhijun; Guo, Jun; Zhao, Xinmin; Zhang, Feng

2016-03-01

Discrete biomolecule-nanoparticle (NP) conjugates play paramount roles in nanofabrication, in which the key is to get the precise molar extinction coefficient of NPs. By making best use of the gift from a specific separation phenomenon of agarose gel electrophoresis (GE), amphiphilic polymer coated NP with exact number of bovine serum albumin (BSA) proteins can be extracted and further experimentally employed to precisely calculate the molar extinction coefficient of the NPs. This method could further benefit the evaluation and extraction of any other dual-component NP-containing bio-conjugates.

Aerosol-assisted chemical vapor deposition of ultra-thin CuOx films as hole transport material for planar perovskite solar cells

NASA Astrophysics Data System (ADS)

Zhang, Zhixin; Chen, Shuqun; Li, Pingping; Li, Hongyi; Wu, Junshu; Hu, Peng; Wang, Jinshu

This paper reports on the fabrication of CuOx films to be used as hole transporting layer (HTL) in CH3NH3PbI3 perovskite solar cells (PSCs). Ultra-thin CuOx coatings were grown onto FTO substrates for the first time via aerosol-assisted chemical vapor deposition (AACVD) of copper acetylacetonate in methanol. After incorporating into the PSCs prepared at ambient air, a highest power conversion efficiency (PCE) of 8.26% with HTL and of 3.34% without HTL were achieved. Our work represents an important step in the development of low-cost CVD technique for fabricating ultra-thin metal oxide functional layers in thin film photovoltaics.

Novel method for the high-throughput processing of slides for the comet assay

PubMed Central

Karbaschi, Mahsa; Cooke, Marcus S.

2014-01-01

Single cell gel electrophoresis (the comet assay), continues to gain popularity as a means of assessing DNA damage. However, the assay's low sample throughput and laborious sample workup procedure are limiting factors to its application. “Scoring”, or individually determining DNA damage levels in 50 cells per treatment, is time-consuming, but with the advent of high-throughput scoring, the limitation is now the ability to process significant numbers of comet slides. We have developed a novel method by which multiple slides may be manipulated, and undergo electrophoresis, in batches of 25 rather than individually and, importantly, retains the use of standard microscope comet slides, which are the assay convention. This decreases assay time by 60%, and benefits from an electrophoresis tank with a substantially smaller footprint, and more uniform orientation of gels during electrophoresis. Our high-throughput variant of the comet assay greatly increases the number of samples analysed, decreases assay time, number of individual slide manipulations, reagent requirements and risk of damage to slides. The compact nature of the electrophoresis tank is of particular benefit to laboratories where bench space is at a premium. This novel approach is a significant advance on the current comet assay procedure. PMID:25425241

Novel method for the high-throughput processing of slides for the comet assay.

PubMed

Karbaschi, Mahsa; Cooke, Marcus S

2014-11-26

Single cell gel electrophoresis (the comet assay), continues to gain popularity as a means of assessing DNA damage. However, the assay's low sample throughput and laborious sample workup procedure are limiting factors to its application. "Scoring", or individually determining DNA damage levels in 50 cells per treatment, is time-consuming, but with the advent of high-throughput scoring, the limitation is now the ability to process significant numbers of comet slides. We have developed a novel method by which multiple slides may be manipulated, and undergo electrophoresis, in batches of 25 rather than individually and, importantly, retains the use of standard microscope comet slides, which are the assay convention. This decreases assay time by 60%, and benefits from an electrophoresis tank with a substantially smaller footprint, and more uniform orientation of gels during electrophoresis. Our high-throughput variant of the comet assay greatly increases the number of samples analysed, decreases assay time, number of individual slide manipulations, reagent requirements and risk of damage to slides. The compact nature of the electrophoresis tank is of particular benefit to laboratories where bench space is at a premium. This novel approach is a significant advance on the current comet assay procedure.

Multi-Layer Coating of Ultrathin Polymer Films on Nanoparticles of Alumina by a Plasma Treatment

DTIC Science & Technology

2001-01-01

Proc. Vol. 635 © 2001 Materials Research Society Multi-Layer Coating of Ultrathin Polymer Films on Nanoparticles of Alumina by a Plasma Treatment Donglu...interconnected organic and inorganic networks results in coatings with a very low permeability for gases and liquids. Hybrid materials are very suitable for... materials consist of a clear alcoholic solution that can easily be processed by classical application techniques such as dipping, spraying, or spin coating

Assessing Mercury and Methylmercury Bioavailability in Sediment Pore Water Using Mercury-Specific Hydrogels

DTIC Science & Technology

2015-06-01

gram AVS acid volatile sulfides BrCl bromium chloride cm centimeter(s) cm2 g-1 square centimeter(s) per gram CVAFS cold vapor atomic...Production The DGT devices used in our experiments consist of three principal components: a diffusive gel, a resin gel, and a membrane. Gel synthesis is...based on the laboratory procedures for the synthesis of polyacrylamide electrophoresis gels (Clarisse and Hintelmann 2006); although, instead of

Ultraviolet-B radiation induced crosslinking improves physical properties of cold- and warm-water fish gelatin gels and films

USDA-ARS?s Scientific Manuscript database

Cold- and warm-water fish gelatin granules were exposed to ultraviolet-B radiation for doses up to 29.7 J/cm2. Solutions and films were prepared from the granules. Gel electrophoresis and refractive index were used to examine changes in molecular weight of the samples. Also, the gel strength and rhe...

Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry

NASA Astrophysics Data System (ADS)

Zhang, Wenyang; Yuan, Zhiwei; Huang, Lulu; Kang, Jie; Jiang, Ruowei; Zhong, Hongying

2016-02-01

Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide.

Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry.

PubMed

Zhang, Wenyang; Yuan, Zhiwei; Huang, Lulu; Kang, Jie; Jiang, Ruowei; Zhong, Hongying

2016-02-11

Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide.

Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry

PubMed Central

Zhang, Wenyang; Yuan, Zhiwei; Huang, Lulu; Kang, Jie; Jiang, Ruowei; Zhong, Hongying

2016-01-01

Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide. PMID:26865351

Alterations in brain cerebral cortex proteome of rabies-infected cat.

PubMed

Kasempimolporn, Songsri; Lumlertdacha, Boonlert; Chulasugandha, Pannipa; Boonchang, Supatsorn; Sitprija, Visith

2014-07-01

Comparative proteome analysis using brain cerebral cortex tissues from cats and dogs infected with/without rabies virus were conducted using both two-dimensional gel-electrophoresis (2-DE) and 2-D fluorescence difference gel- electrophoresis (2D-DIGE) methods. The 2-DE gel images of all samples revealed >1,000 protein spots in each gel. Quantitative intensity analysis revealed the same overall protein pattern in certain regions of the gel, but the rabies-infected brains exhibited more protein spots than the non-infected controls. From approximately 880 protein spots detected by 2D-DIGE, 65 protein spots were increased and 46 were decreased. Eight of these protein spots were randomly selected and annotated by reference to previous known proteome data of rabid dog brains. They were similarly altered in both of the rabies-infected cats and dogs. A more detailed comparison of changes in proteomic profiles of brains between rabid cats and dogs should shed some light on the pathophysiological mechanism of rabies in domestic animals, as most rabies cases have been traceable to or believed to have originated from rabid dogs.

Atomic layer deposition and post-growth thermal annealing of ultrathin MoO3 layers on silicon substrates: Formation of surface nanostructures

NASA Astrophysics Data System (ADS)

Liu, Hongfei; Yang, Ren Bin; Yang, Weifeng; Jin, Yunjiang; Lee, Coryl J. J.

2018-05-01

Ultrathin MoO3 layers have been grown on Si substrates at 120 °C by atomic layer deposition (ALD) using molybdenum hexacarbonyl [Mo(CO)6] and ozone (O3) as the Mo- and O-source precursors, respectively. The ultrathin films were further annealed in air at Tann = 550-750 °C for 15 min. Scanning-electron microscopy, energy-dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy have been employed to evaluate the morphological and elemental properties as well as their evolutions upon annealing of the thin films. They revealed an interfacial SiOx layer in between the MoO3 layer and the Si substrate; this SiOx layer converted into SiO2 during the annealing; and the equivalent thickness of the MoO3 (SiO2) layer decreased (increased) with the increase in Tann. Particles with diameters smaller than 50 nm emerged at Tann = 550 °C and their sizes (density) were reduced (increased) by increasing Tann to 650 °C. A further increase of Tann to 750 °C resulted in telephone-cord-like MoO3 structures, initiated from isolated particles on the surface. These observations have been discussed and interpreted based on temperature-dependent atomic interdiffusions, surface evaporations, and/or melting of MoO3, which shed new light on ALD MoO3 towards its electronic applications.

Purification and characterization of acid trehalase from the yeast suc2 mutant.

PubMed

Mittenbühler, K; Holzer, H

1988-06-15

Acid trehalase was purified from the yeast suc2 deletion mutant. After hydrophobic interaction chromatography, the enzyme could be purified to a single band or peak by a further step of either polyacrylamide gel electrophoresis, gel filtration, or isoelectric focusing. An apparent molecular mass of 218,000 Da was calculated from gel filtration. Polyacrylamide gel electrophoresis of the purified enzyme in the presence of sodium dodecyl sulfate suggested a molecular mass of 216,000 Da. Endoglycosidase H digestion of the purified enzyme resulted after sodium dodecyl sulfate gel electrophoresis in one distinct band at 41,000 Da, representing the mannose-free protein moiety of acid trehalase. The carbohydrate content of the enzyme was 86%. Amino acid analysis indicated 354 residues/molecule of enzyme including 9 cysteine moieties and only 1 methionine. The isoelectric point of the enzyme was estimated by gel electrofocusing to be approximately 4.7. The catalytic activity showed a maximum at pH 4.5. The activity of the enzyme was not inhibited by 10 mM each of HgCl2, EDTA, iodoacetic acid, phenanthrolinium chloride or phenylmethylsulfonyl fluoride. There was no activation by divalent metal ions. The acid trehalase exhibited an apparent Km for trehalose of 4.7 +/- 0.1 mM and a Vmax of 99 mumol of trehalose min-1 X mg-1 at 37 degrees C and pH 4.5. The acid trehalase is located in the vacuoles. The rabbit antiserum raised against acid trehalase exhibited strong cross-reaction with purified invertase. These cross-reactions were removed by affinity chromatography using invertase coupled to CNBr-activated Sepharose 4B. Precipitation of acid trehalase activity was observed with the purified antiserum.

Synthesis, properties and applications of 2D non-graphene materials.

PubMed

Wang, Feng; Wang, Zhenxing; Wang, Qisheng; Wang, Fengmei; Yin, Lei; Xu, Kai; Huang, Yun; He, Jun

2015-07-24

As an emerging class of new materials, two-dimensional (2D) non-graphene materials, including layered and non-layered, and their heterostructures are currently attracting increasing interest due to their promising applications in electronics, optoelectronics and clean energy. In contrast to traditional semiconductors, such as Si, Ge and III-V group materials, 2D materials show significant merits of ultrathin thickness, very high surface-to-volume ratio, and high compatibility with flexible devices. Owing to these unique properties, while scaling down to ultrathin thickness, devices based on these materials as well as artificially synthetic heterostructures exhibit novel and surprising functions and performances. In this review, we aim to provide a summary on the state-of-the-art research activities on 2D non-graphene materials. The scope of the review will cover the preparation of layered and non-layered 2D materials, construction of 2D vertical van der Waals and lateral ultrathin heterostructures, and especially focus on the applications in electronics, optoelectronics and clean energy. Moreover, the review is concluded with some perspectives on the future developments in this field.

Prediction of transmittance spectra for transparent composite electrodes with ultra-thin metal layers

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

Zhao, Zhao; Alford, T. L., E-mail: TA@asu.edu; Khorasani, Arash Elhami

2015-11-28

Recent interest in indium-free transparent composite-electrodes (TCEs) has motivated theoretical and experimental efforts to better understand and enhance their electrical and optical properties. Various tools have been developed to calculate the optical transmittance of multilayer thin-film structures based on the transfer-matrix method. However, the factors that affect the accuracy of these calculations have not been investigated very much. In this study, two sets of TCEs, TiO{sub 2}/Au/TiO{sub 2} and TiO{sub 2}/Ag/TiO{sub 2}, were fabricated to study the factors that affect the accuracy of transmittance predictions. We found that the predicted transmittance can deviate significantly from measured transmittance for TCEs thatmore » have ultra-thin plasmonic metal layers. The ultrathin metal layer in the TCE is typically discontinuous. When light interacts with the metallic islands in this discontinuous layer, localized surface plasmons are generated. This causes extra light absorption, which then leads to the actual transmittance being lower than the predicted transmittance.« less

Photoluminescence and capacitance voltage characterization of GaAs surface passivated by an ultrathin GaN interface control layer

NASA Astrophysics Data System (ADS)

Anantathanasarn, Sanguan; Hasegawa, Hideki

2002-05-01

A novel surface passivation technique for GaAs using an ultrathin GaN interface control layer (GaN ICL) formed by surface nitridation was characterized by ultrahigh vacuum (UHV) photoluminescence (PL) and capacitance-voltage ( C- V) measurements. The PL quantum efficiency was dramatically enhanced after being passivated by the GaN ICL structure, reaching as high as 30 times of the initial clean GaAs surface. Further analysis of PL data was done by the PL surface state spectroscopy (PLS 3) simulation technique. PL and C- V results are in good agreement indicating that ultrathin GaN ICL reduces the gap states and unpins the Fermi level, realizing a wide movement of Fermi level within the midgap region and reduction of the effective surface recombination velocity by a factor of 1/60. GaN layer also introduced a large negative surface fixed charge of about 10 12 cm -2. A further improvement took place by depositing a Si 3N 4 layer on GaN ICL/GaAs structure.

Immunogenic membrane-associated proteins of Mycobacterium tuberculosis revealed by proteomics.

PubMed

Sinha, Sudhir; Kosalai, K; Arora, Shalini; Namane, Abdelkader; Sharma, Pawan; Gaikwad, Anil N; Brodin, Priscille; Cole, Stewart T

2005-07-01

Membrane-associated proteins of Mycobacterium tuberculosis offer a challenge, as well as an opportunity, in the quest for better therapeutic and prophylactic interventions against tuberculosis. The authors have previously reported that extraction with the detergent Triton X-114 (TX-114) is a useful step in proteomic analysis of mycobacterial cell membranes, and detergent-soluble membrane proteins of mycobacteria are potent stimulators of human T cells. In this study 1-D and 2-D gel electrophoresis-based protocols were used for the analysis of proteins in the TX-114 extract of M. tuberculosis membranes. Peptide mass mapping (using MALDI-TOF-MS, matrix assisted laser desorption/ionization time of flight mass spectrometry) of 116 samples led to the identification of 105 proteins, 9 of which were new to the M. tuberculosis proteome. Functional orthologues of 73 of these proteins were also present in Mycobacterium leprae, suggesting their relative importance. Bioinformatics predicted that as many as 73% of the proteins had a hydrophobic disposition. 1-D gel electrophoresis revealed more hydrophobic/transmembrane and basic proteins than 2-D gel electrophoresis. Identified proteins fell into the following major categories: protein synthesis, cell wall biogenesis/architecture and conserved hypotheticals/unknowns. To identify immunodominant proteins of the detergent phase (DP), 14 low-molecular-mass fractions prepared by continuous-elution gel electrophoresis were subjected to T cell activation assays using blood samples from BCG-vaccinated healthy donors from a tuberculosis endemic area. Analysis of the responses (cell proliferation and IFN-gamma production) showed that the immunodominance of certain DP fractions was most probably due to ribosomal proteins, which is consistent with both their specificity for mycobacteria and their abundance. Other membrane-associated proteins, including transmembrane proteins/lipoproteins and ESAT-6, did not appear to contribute significantly to the observed T cell responses.

URPD: a specific product primer design tool

PubMed Central

2012-01-01

Background Polymerase chain reaction (PCR) plays an important role in molecular biology. Primer design fundamentally determines its results. Here, we present a currently available software that is not located in analyzing large sequence but used for a rather straight-forward way of visualizing the primer design process for infrequent users. Findings URPD (yoUR Primer Design), a web-based specific product primer design tool, combines the NCBI Reference Sequences (RefSeq), UCSC In-Silico PCR, memetic algorithm (MA) and genetic algorithm (GA) primer design methods to obtain specific primer sets. A friendly user interface is accomplished by built-in parameter settings. The incorporated smooth pipeline operations effectively guide both occasional and advanced users. URPD contains an automated process, which produces feasible primer pairs that satisfy the specific needs of the experimental design with practical PCR amplifications. Visual virtual gel electrophoresis and in silico PCR provide a simulated PCR environment. The comparison of Practical gel electrophoresis comparison to virtual gel electrophoresis facilitates and verifies the PCR experiment. Wet-laboratory validation proved that the system provides feasible primers. Conclusions URPD is a user-friendly tool that provides specific primer design results. The pipeline design path makes it easy to operate for beginners. URPD also provides a high throughput primer design function. Moreover, the advanced parameter settings assist sophisticated researchers in performing experiential PCR. Several novel functions, such as a nucleotide accession number template sequence input, local and global specificity estimation, primer pair redesign, user-interactive sequence scale selection, and virtual and practical PCR gel electrophoresis discrepancies have been developed and integrated into URPD. The URPD program is implemented in JAVA and freely available at http://bio.kuas.edu.tw/urpd/. PMID:22713312

URPD: a specific product primer design tool.

PubMed

Chuang, Li-Yeh; Cheng, Yu-Huei; Yang, Cheng-Hong

2012-06-19

Polymerase chain reaction (PCR) plays an important role in molecular biology. Primer design fundamentally determines its results. Here, we present a currently available software that is not located in analyzing large sequence but used for a rather straight-forward way of visualizing the primer design process for infrequent users. URPD (yoUR Primer Design), a web-based specific product primer design tool, combines the NCBI Reference Sequences (RefSeq), UCSC In-Silico PCR, memetic algorithm (MA) and genetic algorithm (GA) primer design methods to obtain specific primer sets. A friendly user interface is accomplished by built-in parameter settings. The incorporated smooth pipeline operations effectively guide both occasional and advanced users. URPD contains an automated process, which produces feasible primer pairs that satisfy the specific needs of the experimental design with practical PCR amplifications. Visual virtual gel electrophoresis and in silico PCR provide a simulated PCR environment. The comparison of Practical gel electrophoresis comparison to virtual gel electrophoresis facilitates and verifies the PCR experiment. Wet-laboratory validation proved that the system provides feasible primers. URPD is a user-friendly tool that provides specific primer design results. The pipeline design path makes it easy to operate for beginners. URPD also provides a high throughput primer design function. Moreover, the advanced parameter settings assist sophisticated researchers in performing experiential PCR. Several novel functions, such as a nucleotide accession number template sequence input, local and global specificity estimation, primer pair redesign, user-interactive sequence scale selection, and virtual and practical PCR gel electrophoresis discrepancies have been developed and integrated into URPD. The URPD program is implemented in JAVA and freely available at http://bio.kuas.edu.tw/urpd/.

Design of an ultra-thin absorption layer with magnetic materials based on genetic algorithm at the S band

NASA Astrophysics Data System (ADS)

Wang, Fang; Yang, Xiaoning; Liu, Xiaoning; Niu, Tiaoming; Wang, Jing; Mei, Zhonglei; Jian, Yabin

2018-04-01

In this work, we design an ultra-thin absorption coating at the S band, and the total thickness is less than 2 mm. For incident angle less than 30 degree and the whole S band, the reflection is less than -5 dB. The coating is constructed with 4/3 layers of magnetic material with different thicknesses, which are optimized by using genetic algorithm. Analytic and simulation results confirm the correctness of the design.

Preparation and characterization of the nanoporous ultrathin multilayer films based on molybdenum polyoxometalate (Mo 38) n

NASA Astrophysics Data System (ADS)

Wang, L.; Jiang, M.; Wang, E. B.; Duan, L. Y.; Hao, N.; Lan, Y.; Xu, L.; Li, Z.

2003-11-01

Ultrathin multilayer films of the wheel-shaped molybdenum polyoxometalate cluster (Mo 38) n and poly(allylamine hydrochloride)(PAH) have been prepared by the layer-by-layer (LbL) self-assembly method. The ((Mo 38) n/PAH) m multilayer films have been characterized by X-ray photoelectron spectra (XPS) and atomic force microscopy (AFM). UV-VIS measurements reveal regular film growth with each (Mo 38) n adsorption. The electrochemistry behavior of the film at room temperature was investigated.

Performance enhancement in organic photovoltaic solar cells using iridium (Ir) ultra-thin surface modifier (USM)

NASA Astrophysics Data System (ADS)

Pandey, Rina; Lim, Ju Won; Kim, Jung Hyuk; Angadi, Basavaraj; Choi, Ji Won; Choi, Won Kook

2018-06-01

In this study, Iridium (Ir) metallic layer as an ultra-thin surface modifier (USM) was deposited on ITO coated glass substrate using radio frequency magnetron sputtering for improving the photo-conversion efficiency of organic photovoltaic cells. Ultra-thin Ir acts as a surface modifier replacing the conventional hole transport layer (HTL) PEDOT:PSS in organic photovoltaic (OPV) cells with two different active layers P3HT:PC60BM and PTB7:PC70BM. The Ir USM (1.0 nm) coated on ITO glass substrate showed transmittance of 84.1% and work function of >5.0 eV, which is higher than that of ITO (4.5-4.7 eV). The OPV cells with Ir USM (1.0 nm) exhibits increased power conversion efficiency of 3.70% (for P3HT:PC60BM active layer) and 7.28% (for PTB7:PC70BM active layer) under 100 mW/cm2 illumination (AM 1.5G) which are higher than those of 3.26% and 6.95% for the same OPV cells but with PEDOT:PSS as HTL instead of Ir USM. The results reveal that the chemically stable Ir USM layer could be used as an alternative material for PEDOT:PSS in organic photovoltaic cells.

Counterion dye staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and tryptic gel digestion of stained protein for mass spectrometry.

PubMed

Cong, Wei-Tao; Wang, Xu; Hwang, Sun-Young; Jin, Li-Tai; Choi, Jung-Kap

2012-01-01

A fast and matrix-assisted laser desorption/ionization mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon and ethyl violet, to form an ion-pair complex. The protocol, including fixing, staining, and quick washing steps, can be completed in 1-1.5 h, depending upon gel thickness. It has the sensitivity comparable to the colloidal Coomassie Brilliant Blue G stain using phosphoric acid as a component of staining solution (4-8 ng). The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from mass spectrometry. Considering the speed, sensitivity, and compatibility with mass spectrometry, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches.

Bio-barcode gel assay for microRNA

NASA Astrophysics Data System (ADS)

Lee, Hyojin; Park, Jeong-Eun; Nam, Jwa-Min

2014-02-01

MicroRNA has been identified as a potential biomarker because expression level of microRNA is correlated with various cancers. Its detection at low concentrations would be highly beneficial for cancer diagnosis. Here, we develop a new type of a DNA-modified gold nanoparticle-based bio-barcode assay that uses a conventional gel electrophoresis platform and potassium cyanide chemistry and show this assay can detect microRNA at aM levels without enzymatic amplification. It is also shown that single-base-mismatched microRNA can be differentiated from perfectly matched microRNA and the multiplexed detection of various combinations of microRNA sequences is possible with this approach. Finally, differently expressed microRNA levels are selectively detected from cancer cells using the bio-barcode gel assay, and the results are compared with conventional polymerase chain reaction-based results. The method and results shown herein pave the way for practical use of a conventional gel electrophoresis for detecting biomolecules of interest even at aM level without polymerase chain reaction amplification.

Modeling the Dynamics of Gel Electrophorresis in the High School Classroom

NASA Astrophysics Data System (ADS)

Saucedo, Skyler R.

2013-01-01

Gel electrophoresis, used by geneticists and forensic experts alike, is an immensely popular technique that utilizes an electric field to separate molecules and proteins by size and charge. At the microscopic level, a dye or complex protein like DNA is passed through agarose, a gelatinous three-dimensional matrix of pores and nano-sized tunnels. When forced through a maze of holes, the molecule unravels, forming a long chain, slithering through the field of pores in a process colloquially coined "reputation." As a result, the smaller molecules travel farther through the gel when compared to molecules of larger molecular weight. This highly effective "molecular sieve" provides consistent data and allows scientists to compare similar sequences of DNA base pairs in a routine fashion.2 When performed at the high school level, gel electrophoresis provides students the opportunity to learn about a contemporary lab technique of great scientific relevance. Doing real science certainly excites students and motivates them to learn more.

Electrochemical Corrosion Properties of Commercial Ultra-Thin Copper Foils

NASA Astrophysics Data System (ADS)

Yen, Ming-Hsuan; Liu, Jen-Hsiang; Song, Jenn-Ming; Lin, Shih-Ching

2017-08-01

Ultra-thin electrodeposited Cu foils have been developed for substrate thinning for mobile devices. Considering the corrosion by residual etchants from the lithography process for high-density circuit wiring, this study investigates the microstructural features of ultra-thin electrodeposited Cu foils with a thickness of 3 μm and their electrochemical corrosion performance in CuCl2-based etching solution. X-ray diffraction and electron backscatter diffraction analyses verify that ultra-thin Cu foils exhibit a random texture and equi-axed grains. Polarization curves show that ultra-thin foils exhibit a higher corrosion potential and a lower corrosion current density compared with conventional (220)-oriented foils with fan-like distributed fine-elongated columnar grains. Chronoamperometric results also suggest that ultra-thin foils possess superior corrosion resistance. The passive layer, mainly composed of CuCl and Cu2O, forms and dissolves in sequence during polarization.

Further development of an electroosmotic medium pump system for preparative disk gel electrophoresis.

PubMed

Hayakawa, Mitsuo; Hosogi, Yumiko; Takiguchi, Hisashi; Shiroza, Teruaki; Shibata, Yasuko; Hiratsuka, Koichi; Kiyama-Kishikawa, Michiko; Hamajima, Susumu; Abiko, Yoshimitsu

2003-02-01

A simple and practical 6.8-cm-diameter (36.30-cm(2) cross-sectional-area) preparative disk gel electrophoresis device, based on the design of M. Hayakawa et al. (Anal. Biochem. 288 (2001) 168), in which the elution buffer is driven by an electroosmotic buffer flow through the membrane into the elution chamber from the anode chamber was constructed. We have found that the dialysis membranes employed provide suitable flow rates for the elution buffer, similar to those of an earlier 3.6-cm-diameter device, resulting in the prevention of excess eluate dilution. The efficiency of this device was demonstrated by the fractionation of a bovine serum albumin (BSA) Cohn V fraction into monomer, dimer, and oligomer components using nondenaturing polyacrylamide gel electrophoresis (native-PAGE). The maximum protein concentration of the eluate achieved was 133 mg/ml of BSA monomer, which required a dilution of the eluate for subsequent analytical PAGE performance. As a practical example, the two-dimensional fractionation of soluble dipeptidyl peptidase IV (sDPP IV) from 50 ml fetal bovine serum (3.20 g protein) per gel is presented. The sDPP IV enzyme protein was recovered in a relatively short time, utilizing a 6.5% T native-PAGE and subsequential sodium dodecyl sulfate-PAGE system. This device enhances the possibility of continuous electrophoretic fractionation of complex protein mixtures on a preparative scale. Copyright 2003 Elsevier Science (USA)

Real-time nested multiplex PCR for the detection of herpes simplex virus types 1 and 2 and varicella zoster virus.

PubMed

O'Neill, Hugh J; Wyatt, Dorothy E; Coyle, Peter V; McCaughey, Conall; Mitchell, Frederick

2003-12-01

One hundred forty-nine specimens were tested in a LightCycler nested multiplex polymerase chain reaction (LCnmPCR) for Herpes simplex virus (HSV)1, HSV2, and VZV. Eighty-one were from genitourinary medicine (GUM) patients and the other 68 specimens were from other patients with skin lesions. The results were compared to a conventional multiplex nested PCR (nmPCR) using agarose gel electrophoresis. Twenty-five specimens were positive in both assays for HSV1 and 29 were positive for VZV. For HSV2 there were 27 positive in the LCnmPCR and 26 positive in the nmPCR assay. The melting temperatures (Tms) of each target were different with a mean of 84.75 degrees C for HSV1, 88.57 degrees C for HSV2, and 83.62 degrees C for VZV. The melting curves of positive specimens directly overlaid the melting curves of the positive controls in the assay. The LCnmPCR assay is a convenient alternative to conventional PCR using agarose gel electrophoresis. It improves specimen turnaround time by eliminating the need for gel electrophoresis, transillumination, and gel photography. It also shows increased sensitivity for HSV2 over our standard assay. This LCnmPCR reduces further the possibility of amplicon contamination with nested PCR protocols. Copyright 2003 Wiley-Liss, Inc.

Colored ultra-thin hybrid photovoltaics with high quantum efficiency for decorative PV applications (Presentation Recording)

NASA Astrophysics Data System (ADS)

Guo, L. Jay

2015-10-01

This talk will describe an approach to create architecturally compatible and decorative thin-film-based hybrid photovoltaics [1]. Most current solar panels are fabricated via complex processes using expensive semiconductor materials, and they are rigid and heavy with a dull, black appearance. As a result of their non-aesthetic appearance and weight, they are primarily installed on rooftops to minimize their negative impact on building appearance. Recently we introduced dual-function solar cells based on ultra-thin dopant-free amorphous silicon embedded in an optical cavity that not only efficiently extract the photogenerated carriers but also display distinctive colors with the desired angle-insensitive appearances [1,2]. The angle-insensitive behavior is the result of an interesting phase cancellation effect in the optical cavity with respect to angle of light propagation [3]. In order to produce the desired optical effect, the semiconductor layer should be ultra-thin and the traditional doped layers need to be eliminated. We adopted the approach of employing charge transport/blocking layers used in organic solar cells to meet this demand. We showed that the ultra-thin (6 to 31 nm) undoped amorphous silicon/organic hybrid solar cell can transmit desired wavelength of light and that most of the absorbed photons in the undoped a-Si layer contributed to the extracted electric charges. This is because the a-Si layer thickness is smaller than the charge diffusion length, therefore the electron-hole recombination is strongly suppressed in such ultra-thin layer. Reflective colored PVs can be made in a similar fashion. Light-energy-harvesting colored signage was demonstrated. Furthermore, a cascaded photovoltaics scheme based on tunable spectrum splitting can be employed to increase power efficiency by absorbing a broader band of light energy. Our work provides a guideline for optimizing a photoactive layer thickness in high efficiency hybrid PV design, which can be adopted by other material systems as well. Based on these understandings, we have also developed colored perovskite PV by integrating an optical cavity with the perovskite semiconductors [4]. The principle and experimental results will be presented. 1. J. Y. Lee, K. T. Lee, S.Y. Seo, L. J. Guo, "Decorative power generating panels creating angle insensitive transmissive colors," Sci. Rep. 4, 4192, 2014. 2. K. T. Lee, J.Y. Lee, S.-Y. Seo, and L. J. Guo, "Colored ultra-thin hybrid photovoltaics with high quantum efficiency," Light: Science and Applications, 3, e215, 2014. 3. K. T. Lee, S.-Y. Seo, J.Y. Lee, and L. J. Guo, "Ultrathin metal-semiconductor-metal resonator for angle invariant visible band transmission filters," Appl. Phys. Lett. 104, 231112, (2014); and "Strong resonance effect in a lossy medium-based optical cavity for angle robust spectrum filters," Adv. Mater, 26, 6324-6328, 2014. 4. K. T. Lee, M. Fukuda, L. J. Guo, "Colored, see-through perovskite solar cells employing an optical cavity," Submitted, 2015

Proteomic analysis of albumin and globulin fractions of pea (Pisum sativum L.) seeds.

PubMed

Dziuba, Jerzy; Szerszunowicz, Iwona; Nałęcz, Dorota; Dziuba, Marta

2014-01-01

Proteomic analysis is emerging as a highly useful tool in food research, including studies of food allergies. Two-dimensional gel electrophoresis involving isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis is the most effective method of separating hundreds or even thousands of proteins. In this study, albumin and globulin tractions of pea seeds cv. Ramrod were subjected to proteomic analysis. Selected potentially alergenic proteins were identified based on their molecular weights and isoelectric points. Pea seeds (Pisum sativum L.) cv. Ramrod harvested over a period of two years (Plant Breeding Station in Piaski-Szelejewo) were used in the experiment. The isolated albumins, globulins and legumin and vicilin fractions of globulins were separated by two-dimensional gel electrophoresis. Proteomic images were analysed in the ImageMaster 2D Platinum program with the use of algorithms from the Melanie application. The relative content, isoelectric points and molecular weights were computed for all identified proteins. Electrophoregrams were analysed by matching spot positions from three independent replications. The proteomes of albumins, globulins and legumin and vicilin fractions of globulins produced up to several hundred spots (proteins). Spots most characteristic of a given fraction were identified by computer analysis and spot matching. The albumin proteome accumulated spots of relatively high intensity over a broad range of pi values of ~4.2-8.1 in 3 molecular weight (MW) ranges: I - high molecular-weight albumins with MW of ~50-110 kDa, II - average molecular-weight albumins with MW of ~20-35 kDa, and III - low molecular-weight albumins with MW of ~13-17 kDa. 2D gel electrophoregrams revealed the presence of 81 characteristic spots, including 24 characteristic of legumin and 14 - of vicilin. Two-dimensional gel electrophoresis proved to be a useful tool for identifying pea proteins. Patterns of spots with similar isoelectric points and different molecular weights or spots with different isoelectric points and similar molecular weights play an important role in proteome analysis. The regions characteristic of albumin, globulin and legumin and vicilin fractions of globulin with typical MW and pi values were identified as the results of performed 2D electrophoretic separations of pea proteins. 2D gel electrophoresis of albumins and the vicilin fraction of globulins revealed the presence of 4 and 2 spots, respectively, representing potentially allergenic proteins. They probably corresponded to vicilin fragments synthesized during post-translational modification of the analysed protein.

Separation and characterization of high-density lipoprotein subpopulations by gel permeation chromatography.

PubMed

Clifton, P M; MacKinnon, A M; Barter, P J

1987-02-20

High-density lipoproteins (HDL) contain at least five distinct subpopulations when analyzed by gradient gel electrophoresis. This report represents the first description of a simple technique for isolating these subpopulations of HDL in quantities sufficient to enable characterization in terms of particle size, apolipoprotein AI and apolipoprotein AII content and chemical composition. Lipoproteins were separated and subfractionated on a column of Superose 6B using a fast protein liquid chromatography system. Five normal subjects were studied: HDL2b and HDL3a were isolated as essentially single subpopulations from all subjects, while HDL2a could be isolated from only three of the subjects. HDL3b was isolated in a relatively impure form (70%) from all subjects. Identical subpopulations were identified in each subject by gradient gel electrophoresis of unseparated HDL.

A rapid method to visualize von willebrand factor multimers by using agarose gel electrophoresis, immunolocalization and luminographic detection.

PubMed

Krizek, D R; Rick, M E

2000-03-15

A highly sensitive and rapid clinical method for the visualization of the multimeric structure of von Willebrand Factor in plasma and platelets is described. The method utilizes submerged horizontal agarose gel electrophoresis, followed by transfer of the von Willebrand Factor onto a polyvinylidine fluoride membrane, and immunolocalization and luminographic visualization of the von Willebrand Factor multimeric pattern. This method distinguishes type 1 from types 2A and 2B von Willebrand disease, allowing timely evaluation and classification of von Willebrand Factor in patient plasma. It also allows visualization of the unusually high molecular weight multimers present in platelets. There are several major advantages to this method including rapid processing, simplicity of gel preparation, high sensitivity to low concentrations of von Willebrand Factor, and elimination of radioactivity.

Nanoassembled thin film gas sensors. III. Sensitive detection of amine odors using TiO2/poly(acrylic acid) ultrathin film quartz crystal microbalance sensors.

PubMed

Lee, Seung-Woo; Takahara, Naoki; Korposh, Sergiy; Yang, Do-Hyeon; Toko, Kiyoshi; Kunitake, Toyoki

2010-03-15

Quartz crystal microbalance (QCM) gas sensors based on the alternate adsorption of TiO(2) and polyacrilic acid (PAA) were developed for the sensitive detection of amine odors. Individual TiO(2) gel layers could be regularly assembled with a thickness of approximately 0.3 nm by the gas-phase surface sol-gel process (GSSG). The thickness of the poly(acrylic acid) (PAA) layer is dependent on its molecular weight, showing different thicknesses of approximately 0.4 nm for PAA(25) (Mw 250,000) and 0.6-0.8 nm for PAA(400) (Mw 4,000,000). The QCM sensors showed a linear response to ammonia in the concentration range 0.3-15 ppm, depending on the deposition cycle of the alternate TiO(2)/PAA layer. The ammonia binding is based on the acid-base interaction to the free carboxylic acid groups of PAA and the limit of detection (LOD) of the 20-cycle TiO(2)/PAA(400) film was estimated to be 0.1 ppm when exposed to ammonia. The sensor response was very fast and stable in a wide relative humidity (rH) range of 30-70%, showing almost the same frequency changes at a given concentration of ammonia. Sensitivity to n-butylamine and ammonia was higher than to pyridine, which is owing to the difference of molecular weight and basicity of the amine analytes. The alternate TiO(2)/PAA(400) films have a highly effective ability to capture amine odors, and the ambient ammonia concentration of 15 ppm could be condensed up to approximately 20,000 ppm inside the films.

Ultrathin metal-semiconductor-metal resonator for angle invariant visible band transmission filters

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

Lee, Kyu-Tae; Seo, Sungyong; Yong Lee, Jae

We present transmission visible wavelength filters based on strong interference behaviors in an ultrathin semiconductor material between two metal layers. The proposed devices were fabricated on 2 cm × 2 cm glass substrate, and the transmission characteristics show good agreement with the design. Due to a significantly reduced light propagation phase change associated with the ultrathin semiconductor layer and the compensation in phase shift of light reflecting from the metal surface, the filters show an angle insensitive performance up to ±70°, thus, addressing one of the key challenges facing the previously reported photonic and plasmonic color filters. This principle, described in this paper, canmore » have potential for diverse applications ranging from color display devices to the image sensors.« less

Evaluation of capillary zone electrophoresis for the determination of protein composition in therapeutic immunoglobulins and human albumins.

PubMed

Christians, Stefan; van Treel, Nadine Denise; Bieniara, Gabriele; Eulig-Wien, Annika; Hanschmann, Kay-Martin; Giess, Siegfried

2016-07-01

Capillary zone electrophoresis (CZE) provides an alternative means of separating native proteins on the basis of their inherent electrophoretic mobilities. The major advantage of CZE is the quantification by UV detection, circumventing the drawbacks of staining and densitometry in the case of gel electrophoresis methods. The data of this validation study showed that CZE is a reliable assay for the determination of protein composition in therapeutic preparations of human albumin and human polyclonal immunoglobulins. Data obtained by CZE are in line with "historical" data obtained by the compendial method, provided that peak integration is performed without time correction. The focus here was to establish a rapid and reliable test to substitute the current gel based zone electrophoresis techniques for the control of protein composition of human immunoglobulins or albumins in the European Pharmacopoeia. We believe that the more advanced and modern CZE method described here is a very good alternative to the procedures currently described in the relevant monographs. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

Improved stability and electrophoretic properties of preformed fluorescent cationic dye-DNA complexes in a taps-tetrapentylammonium buffer in agarose slab gels.

PubMed

Zeng, Z; Clark, S M; Mathies, R A; Glazer, A N

1997-10-01

High-resolution capillary electrophoresis sizing of preformed complexes of bis-intercalating fluorescent dyes with double-stranded DNA has been demonstrated using hydroxyethylcellulose and 3-[tris-(hydroxymethyl) methylamino]-1-propanesulfonic acid-tetrapentylammonium (Taps-NPe+4) buffers (S. M. Clark and R. A. Mathies, Anal. Chem. 69, 1355-1363, 1997). Such capillary electrophoresis separations were unattainable in conventional buffers containing other cations such as Tris+, Na+, and NH+4. We report here the behavior of preformed double-stranded DNA-dye complexes on agarose slab gel electrophoresis in 40 mM Taps-NPe+4, 1 mM H2EDTA, pH 8.2. Upon electrophoresis in this buffer (a) complexes formed at DNA base pairs:dye ratios ranging from 100:1 to 5:1 show the same mobility; (b) the half-lives of DNA-dye complexes with monointercalators are two- to threefold longer than those in commonly used Tris buffers; (c) there is little dye transfer between labeled and unlabeled DNA molecules; and (d) precise two-color sizing of preformed restriction fragment-dye complexes with fluorescent bisintercalators is achieved.

Detection of glycoproteins in the Acanthamoeba plasma membrane

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

Paatero, G.I.L.; Gahmberg, C.G.

1988-11-01

In the present study the authors have shown that glycoproteins are present in the plasma membrane of Acanthamoeba castellanii by utilizing different radioactive labeling techniques. Plasma membrane proteins in the amoeba were iodinated by {sup 125}I-lactoperoxidase labeling and the solubilized radiolabeled glycoproteins were separated by lectin-Sepharose affinity chromatography followed by polyacrylamide gel electrophoresis. The periodate/NaB{sup 3}H{sub 4} and galactose oxidase/NaB{sup 3}H{sub 4} labeling techniques were used for labeling of surface carbohydrates in the amoeba. Several surface-labeled glycoproteins were observed in addition to a diffusely labeled region with M{sub r} of 55,000-75,000 seen on electrophoresis, which could represent glycolipids. The presencemore » of glycoproteins in the plasma membrane of Acanthamoeba castellanii was confirmed by metabolic labeling with ({sup 35}S)methionine followed by lectin-Sepharose affinity chromatography and polyacrylamide gel electrophoresis.« less

Electrophoretic Mobility Shift Assay (EMSA) for Detecting Protein-Nucleic Acid Interactions

PubMed Central

Hellman, Lance M.; Fried, Michael G.

2009-01-01

The gel electrophoresis mobility shift assay (EMSA) is used to detect protein complexes with nucleic acids. It is the core technology underlying a wide range of qualitative and quantitative analyses for the characterization of interacting systems. In the classical assay, solutions of protein and nucleic acid are combined and the resulting mixtures are subjected to electrophoresis under native conditions through polyacrylamide or agarose gel. After electrophoresis, the distribution of species containing nucleic acid is determined, usually by autoradiography of 32P-labeled nucleic acid. In general, protein-nucleic acid complexes migrate more slowly than the corresponding free nucleic acid. In this article, we identify the most important factors that determine the stabilities and electrophoretic mobilities of complexes under assay conditions. A representative protocol is provided and commonly used variants are discussed. Expected outcomes are briefly described. References to extensions of the method and a troubleshooting guide are provided. PMID:17703195

Purification of 6-phosphogluconate dehydrogenase from parsley (Petroselinum hortense) leaves and investigation of some kinetic properties.

PubMed

Demir, Hülya; Ciftçi, Mehmet; Küfrevioğlu, O Irfan

2003-02-01

In this study, 6-phosphogluconate dehydrogenase (E.C.1.1.44; 6PGD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps that are preparation of homogenate ammonium sulfate fractionation and on DEAE-Sephadex A50 ion exchange. The enzyme was obtained with a yield of 49% and had a specific activity of 18.3 U (mg proteins)(-1) (Lehninger, A.L.; Nelson, D.L.; Cox, M.M. Principles of Biochemistry, 2nd Ed.; Worth Publishers Inc.: N.Y., 2000, 558-560). The overall purification was about 339-fold. A temperature of +4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method at 340 mn. In order to control the purification of the enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 97.5 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a subunit molecular weight of 24.1 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found as 8.0, 8.0, and 50 degrees C, respectively. In addition, KM and Vmax values for NADP+ and G6-P at optimum pH and 25 degrees C were determined by means of Lineweaver-Burk plots.

A brief review of other notable protein detection methods on acrylamide gels.

PubMed

Kurien, Biji T; Scofield, R Hal

2012-01-01

Several methods have been described to stain proteins analyzed on acrylamide gels. These include ultrasensitive protein detection in one-dimensional and two-dimensional gel electrophoresis using a fluorescent product from the fungus Epicoccum nigrum; a fluorescence-based Coomassie Blue protein staining; visualization of proteins in acrylamide gels using ultraviolet illumination; fluorescence visualization of proteins in sodium dodecyl sulfate-polyacrylamide gels using environmentally benign, nonfixative, saline solution; and increasing the sensitivity four- to sixfold for detecting trace proteins in dye or silver stained polyacrylamide gels using polyethylene glycol 6000. All these methods are reviewed briefly in this chapter.

Ultra-thin L1{sub 0}-FePt for perpendicular anisotropy L1{sub 0}-FePt/Ag/[Co/Pd]{sub 30} pseudo spin valves

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

Ho, Pin; Chow, Gan Moog; Chen, Jing-Sheng, E-mail: msecj@nus.edu.sg

2014-05-07

Perpendicular anisotropy L1{sub 0}-FePt/Ag/[Co/Pd]{sub 30} pseudo spin valves (PSVs) with ultra-thin L1{sub 0}-FePt alloy free layer possessing high anisotropy and thermal stability have been fabricated and studied. The thickness of the L1{sub 0}-FePt layer was varied between 2 and 4 nm. The PSV became increasingly decoupled with reduced L1{sub 0}-FePt thickness due to the larger difference between the coercivity of the L1{sub 0}-FePt and [Co/Pd]{sub 30} films. The PSV with an ultra-thin L1{sub 0}-FePt free layer of 2 nm displayed a high K{sub u} of 2.21 × 10{sup 7} ergs/cm{sup 3}, high thermal stability of 84 and a largest giant magnetoresistance of 0.54%.

Improved DC and RF performance of InAlAs/InGaAs InP based HEMTs using ultra-thin 15 nm ALD-Al2O3 surface passivation

NASA Astrophysics Data System (ADS)

Asif, Muhammad; Chen, Chen; Peng, Ding; Xi, Wang; Zhi, Jin

2018-04-01

Owing to the great influence of surface passivation on DC and RF performance of InP-based HEMTs, the DC and RF performance of InAlAs/InGaAs InP HEMTs were studied before and after passivation, using an ultra-thin 15 nm atomic layer deposition Al2O3 layer. Increase in Cgs and Cgd was significantly limited by scaling the thickness of the Al2O3 layer. For verification, an analytical small-signal equivalent circuit model was developed. A significant increase in maximum transconductance (gm) up to 1150 mS/mm, drain current (IDS) up to 820 mA/mm and fmax up to 369.7 GHz was observed, after passivation. Good agreement was obtained between the measured and the simulated results. This shows that the RF performance of InP-based HEMTs can be improved by using an ultra-thin ALD-Al2O3 surface passivation.

Ultrathin Lutetium Oxide Film as an Epitaxial Hole-Blocking Layer for Crystalline Bismuth Vanadate Water Splitting Photoanodes

DOE PAGES

Zhang, Wenrui; Yan, Danhua; Tong, Xiao; ...

2018-01-08

Here a novel ultrathin lutetium oxide (Lu 2O 3) interlayer is integrated with crystalline bismuth vanadate (BiVO4) thin film photoanodes to facilitate carrier transport through atomic-scale interface control. The epitaxial Lu 2O 32O 3

Antiferromagnetic exchange and magnetoresistance enhancement in ultrathin Co-Re sandwiches

NASA Astrophysics Data System (ADS)

Freitas, P. P.; Melo, L. V.; Trindade, I.; From, M.

1992-10-01

Co-Re ultrathin sandwiches were prepared that show antiferromagnetic coupling and enhanced saturation magnetoresistance for Re spacer thicknesses below 9 Å. A field of 2.5 kOe is needed to saturate the antiferromagnetically coupled Co layers. These results are similar to those found in Co-Re superlattices.

Electronic structure evolution in doping of fullerene (C{sub 60}) by ultra-thin layer molybdenum trioxide

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

Wang, Chenggong; Wang, Congcong; Kauppi, John

2015-08-28

Ultra-thin layer molybdenum oxide doping of fullerene has been investigated using ultraviolet photoemission spectroscopy (UPS) and X-ray photoemission spectroscopy (XPS). The highest occupied molecular orbital (HOMO) can be observed directly with UPS. It is observed that the Fermi level position in fullerene is modified by ultra-thin-layer molybdenum oxide doping, and the HOMO onset is shifted to less than 1.3 eV below the Fermi level. The XPS results indicate that charge transfer was observed from the C{sub 60} to MoO{sub x} and Mo{sup 6+} oxides is the basis as hole dopants.

Improved method for identification of low abundance proteins using 2D-gel electrophoresis, MALDI-TOF and TOF/TOF

EPA Science Inventory

Introduction: Differential protein expression studies have been routinely performed in our laboratory to determine the health effects of environmentally-important chemicals. In this abstract, improvements in the in-gel protein digestion, MALDI plate spotting and data acquisition...

Hybrid slab-microchannel gel electrophoresis system

DOEpatents

Balch, Joseph W.; Carrano, Anthony V.; Davidson, James C.; Koo, Jackson C.

1998-01-01

A hybrid slab-microchannel gel electrophoresis system. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate.

Purification and characterization of a hygromycin B phosphotransferase from Streptomyces hygroscopicus.

PubMed

Zalacain, M; Pardo, J M; Jiménez, A

1987-01-15

A hygromycin B phosphotransferase activity from Streptomyces hygroscopicus has been highly purified by ammonium sulphate fractionation followed by affinity column chromatography through Sepharose-6B-hygromycin-B. The combined active fractions showed a single protein band (41 kDa) when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. When gel electrophoresis was performed under non-denaturing conditions, the single protein band promoted in situ phosphorylation of hygromycin B, indicating that this protein corresponded to the purified hygromycin B phosphotransferase. The enzyme has been purified 236-fold and approximate Km values of 0.56 microM for hygromycin B and ATP, respectively, were deduced.

Contribution of PCR Denaturing Gradient Gel Electrophoresis Combined with Mixed Chromatogram Software Separation for Complex Urinary Sample Analysis.

PubMed

Kotásková, Iva; Mališová, Barbora; Obručová, Hana; Holá, Veronika; Peroutková, Tereza; Růžička, Filip; Freiberger, Tomáš

2017-01-01

Complex samples are a challenge for sequencing-based broad-range diagnostics. We analysed 19 urinary catheter, ureteral Double-J catheter, and urine samples using 3 methodological approaches. Out of the total 84 operational taxonomic units, 37, 61, and 88% were identified by culture, PCR-DGGE-SS (PCR denaturing gradient gel electrophoresis followed by Sanger sequencing), and PCR-DGGE-RM (PCR- DGGE combined with software chromatogram separation by RipSeq Mixed tool), respectively. The latter approach was shown to be an efficient tool to complement culture in complex sample assessment. © 2017 S. Karger AG, Basel.

The use of the 2-aminobenzoic acid tag for oligosaccharide gel electrophoresis.

PubMed

Huang, Z; Prickett, T; Potts, M; Helm, R F

2000-08-18

Gel electrophoresis of fluorophore labeled saccharides provides a rapid and reliable method to screen enzymatic and/or chemical treatments of polysaccharides and glycoconjugates, as well as a sensitive and efficient microscale method to separate and purify oligosaccharides for further analysis. A simple and inexpensive method of derivatization and analysis using 2-aminobenzoic acid (anthranilic acid, AA) is described and applied to the extracellular polysaccharide released by the desiccation tolerant cyanobacterium Nostoc commune DRH-1. The results of these analyses suggest a possible protective functionality of two pendent groups, as well as a potential relationship between these groups and the desiccation tolerance of the organism.

Separation and characterization of needle and xylem maritime pine proteins.

PubMed

Costa, P; Pionneau, C; Bauw, G; Dubos, C; Bahrmann, N; Kremer, A; Frigerio, J M; Plomion, C

1999-01-01

Two-dimensional gel electrophoresis (2-DE) and image analysis are currently used for proteome analysis in maritime pine (Pinus pinaster Ait.). This study presents a database of expressed proteins extracted from needles and xylem, two important tissues for growth and wood formation. Electrophoresis was carried out by isoelectric focusing (IEF) in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second. Silver staining made it possible to detect an average of 900 and 600 spots on 2-DE gels from needles and xylem, respectively. A total of 28 xylem and 35 needle proteins were characterized by internal peptide microsequencing. Out of these 63 proteins, 57 (90%) could be identified based on amino acid similarity with known proteins, of which 24 (42%) have already been described in conifers. Overall comparison of both tissues indicated that 29% and 36% of the spots were specific to xylem and needles, respectively, while the other spots were of identical molecular weight and isoelectric point. The homology of spot location in 2-DE patterns was further validated by sequence analysis of proteins present in both tissues. A proteomic database of maritime pine is accessible on the internet (http://www.pierroton.inra.fr/genetics/2D/).

Automated Parallel Capillary Electrophoretic System

DOEpatents

Li, Qingbo; Kane, Thomas E.; Liu, Changsheng; Sonnenschein, Bernard; Sharer, Michael V.; Kernan, John R.

2000-02-22

An automated electrophoretic system is disclosed. The system employs a capillary cartridge having a plurality of capillary tubes. The cartridge has a first array of capillary ends projecting from one side of a plate. The first array of capillary ends are spaced apart in substantially the same manner as the wells of a microtitre tray of standard size. This allows one to simultaneously perform capillary electrophoresis on samples present in each of the wells of the tray. The system includes a stacked, dual carousel arrangement to eliminate cross-contamination resulting from reuse of the same buffer tray on consecutive executions from electrophoresis. The system also has a gel delivery module containing a gel syringe/a stepper motor or a high pressure chamber with a pump to quickly and uniformly deliver gel through the capillary tubes. The system further includes a multi-wavelength beam generator to generate a laser beam which produces a beam with a wide range of wavelengths. An off-line capillary reconditioner thoroughly cleans a capillary cartridge to enable simultaneous execution of electrophoresis with another capillary cartridge. The streamlined nature of the off-line capillary reconditioner offers the advantage of increased system throughput with a minimal increase in system cost.

Electrphoretic Sample Excitation Light Assembly.

DOEpatents

Li, Qingbo; Liu, Changsheng

2002-04-02

An automated electrophoretic system is disclosed. The system employs a capillary cartridge having a plurality of capillary tubes. The cartridge has a first array of capillary ends projecting from one side of a plate. The first array of capillary ends are spaced apart in substantially the same manner as the wells of a microtitre tray of standard size. This allows one to simultaneously perform capillary electrophoresis on samples present in each of the wells of the tray. The system includes a stacked, dual carrousel arrangement to eliminate cross-contamination resulting from reuse of the same buffer tray on consecutive executions from electrophoresis. The system also has a gel delivery module containing a gel syringe/a stepper motor or a high pressure chamber with a pump to quickly and uniformly deliver gel through the capillary tubes. The system further includes a multi-wavelength beam generator to generate a laser beam which produces a beam with a wide range of wavelengths. An off-line capillary reconditioner thoroughly cleans a capillary cartridge to enable simultaneous execution of electrophoresis with another capillary cartridge. The streamlined nature of the off-line capillary reconditioner offers the advantage of increased system throughput with a minimal increase in system cost.

Motorized Positioning Apparatus Having Coaxial Carrousels.

DOEpatents

Li, Qingbo; Kane, Thomas E.; Liu, Changsheng; Sonnenschein, Bernard; Sharer, Michael V.; Kernan, John R.

2002-04-02

An automated electrophoretic system is disclosed. The system employs a capillary cartridge having a plurality of capillary tubes. The cartridge has a first array of capillary ends projecting from one side of a plate. The first array of capillary ends are spaced apart in substantially the same manner as the wells of a microtitre tray of standard size. This allows one to simultaneously perform capillary electrophoresis on samples present in each of the wells of the tray. The system includes a stacked, dual carrousel arrangement to eliminate cross-contamination resulting from reuse of the same buffer tray on consecutive executions from electrophoresis. The system also has a gel delivery module containing a gel syringe/a stepper motor or a high pressure chamber with a pump to quickly and uniformly deliver gel through the capillary tubes. The system further includes a multi-wavelength beam generator to generate a laser beam which produces a beam with a wide range of wavelengths. An off-line capillary reconditioner thoroughly cleans a capillary cartridge to enable simultaneous execution of electrophoresis with another capillary cartridge. The streamlined nature of the off-line capillary reconditioner offers the advantage of increased system throughput with a minimal increase in system cost.

Fluid Delivery System For Capillary Electrophoretic Applications.

DOEpatents

Li, Qingbo; Liu, Changsheng; Kane, Thomas E.; Kernan, John R.; Sonnenschein, Bernard; Sharer, Michael V.

2002-04-23

An automated electrophoretic system is disclosed. The system employs a capillary cartridge having a plurality of capillary tubes. The cartridge has a first array of capillary ends projecting from one side of a plate. The first array of capillary ends are spaced apart in substantially the same manner as the wells of a microtitre tray of standard size. This allows one to simultaneously perform capillary electrophoresis on samples present in each of the wells of the tray. The system includes a stacked, dual carrousel arrangement to eliminate cross-contamination resulting from reuse of the same buffer tray on consecutive executions from electrophoresis. The system also has a gel delivery module containing a gel syringe/a stepper motor or a high pressure chamber with a pump to quickly and uniformly deliver gel through the capillary tubes. The system further includes a multi-wavelength beam generator to generate a laser beam which produces a beam with a wide range of wavelengths. An off-line capillary reconditioner thoroughly cleans a capillary cartridge to enable simultaneous execution of electrophoresis with another capillary cartridge. The streamlined nature of the off-line capillary reconditioner offers the advantage of increased system throughput with a minimal increase in system cost.

The significance of gtf genes in caries expression: a rapid identification of Streptococcus mutans from dental plaque of child patients.

PubMed

Mishra, Apurva; Pandey, Ramesh K; Manickam, Natesan

2015-01-01

Rapid phylogenetic and functional gene (gtfB) identification of S. mutans from the dental plaque derived from children. Dental plaque collected from fifteen patients of age group 7-12 underwent centrifugation followed by genomic DNA extraction for S. mutans. Genomic DNA was processed with S. mutans specific primers in suitable PCR condtions for phylogenetic and functional gene (gtfB) identification. The yield and results were confirmed by agarose gel electrophoresis. 1% agarose gel electrophoresis depicts the positive PCR amplification at 1,485 bp when compared with standard 1 kbp indicating the presence of S. mutans in the test sample. Another PCR reaction was set using gtfB primers specific for S. mutans for functional gene identification. 1.2% agarose gel electrophoresis was done and a positive amplication was observed at 192 bp when compared to 100 bp standards. With the advancement in molecular biology techniques, PCR based identification and quantification of the bacterial load can be done within hours using species-specific primers and DNA probes. Thus, this technique may reduce the laboratory time spend in conventional culture methods, reduces the possibility of colony identification errors and is more sensitive to culture techniques.

Polymerase chain reaction for detection of invasive Shigella flexneri in food.

PubMed

Lampel, K A; Jagow, J A; Trucksess, M; Hill, W E

1990-06-01

The polymerase chain reaction (PCR) was used to amplify a 760-base-pair (bp) fragment with the 220-kbp invasive plasmids of enteroinvasive Escherichia coli, Shigella flexneri, Shigella dysenteriae, Shigella boydii, and Shigella sonnei as templates. This PCR product was easily detected by agarose gel electrophoresis. A 210-bp AccI-PstI fragment lying within the amplified region was used as a probe in Southern hybridization blots and showed that the PCR-generated product was derived from the invasive plasmid. The application of PCR as a rapid method to detect enteroinvasive bacteria in foods was tested by inoculating lettuce with 10(4) S. flexneri cells per g in shigella broth base. Plasmid DNA was isolated from cultures of inoculated and uninoculated lettuce in broth after 0, 4, and 24 h of incubation. With the PCR, the 760-bp fragment was generated only from lettuce inoculated with S. flexneri, as shown by gel electrophoresis and confirmed both by Southern blotting and by nucleotide sequencing of the amplified region. Because the isolation of plasmid DNA, the performance of PCR, and gel electrophoresis all can be completed in 6 to 7 h, invasive enteric bacteria can be detected in less than 1 day.

Genotoxic potential of bee venom (Apis Mellifera) on human peripheral blood lymphocytes in vitro using single cell gel electrophoresis assay.

PubMed

Gajski, Goran; Garaj-Vrhovac, Vera

2008-09-01

Bee venom (BV) has been known to have therapeutic applications in traditional medicine to treat variety of diseases. It is also known that bee venom possesses anti-inflammatory and anticancer effects and that it can inhibit proliferation and induces apoptosis in cancer cells, but there is lack of information regarding genotoxicity of whole bee venom on normal human cells. In the present study, peripheral blood human lymphocytes from healthy donor were exposed in vitro to different concentration (5, 10, 25, 50 and 100 micro g/mL) of whole bee venom at different time periods (1, 6 and 24 hours). The single cell gel electrophoresis (SCGE) assay was used to evaluate the genotoxicity towards human cells. Results showed statistically significant increase in DNA damage caused in BV treated human lymphocytes compared to corresponding control cells for the tail length and tail moment. These results show that the extent of DNA damage, determined by the use of single cell gel electrophoresis is time and dose dependent. Based on the results it is clear that whole bee venom induces DNA damage and has genotoxic potential on human peripheral blood lymphocytes in vitro.

The changes of serum proteome and tissular pathology in mouse induced by botulinum toxin E injection.

PubMed

Wang, J F; Mao, X Y; Zhao, C

2014-01-01

The experiment were performed to investigate the poisoning-related proteins and main pathological changes after mouse suffered from injection of botulinum toxin serotype E. Dose of 0.75 LD50 botulinum toxin serotype E per mice were administrated by intraperitoneal injection. Survival mouse were picked as experimental group. The blood were collected from orbital blood and serum sample was separated by centrifugation. The heart, liver, spleen, lung, kidney were fixed in 10 % neutral buffered formalin and then developed paraffin sections. Serum protein components were analyzed by SDS-PAGE gel electrophoresis coupled with 2-DE SDS-PAGE gel electrophoresis. Differentially expressed proteins were analyzed by PDQUest8.0 software and subjected to ion trap mass spectrometry equipped with a high performance liquid chromatography system. The observation of pathological section showed that heart, liver, spleen, lung, kidney exhibited pathological changes in different degree, especially in heart, liver and lung tissues. Heart muscle tissue display serious inflammatory response, heart muscle fiber compulsively expanded and filled with erythrocyte and inflammatory exudates, some heart muscle fiber ruptured, even necrosis; hepatic cell in edge of liver occur apoptosis and some hepatic cell have disintegrated, and even died; pulmonary alveoli broken and partial vein filled with blood. Serum proteins component present a significant changes between control serum and botulism in 24 h by SDS-PAGE gel electrophoresis and 2-DE-SDS-PAGE gel electrophoresis. Twenty differentially expressed protein spots were observed in 2-DE profiles, in which 14 protein spots were undetectable in serum proteome under botulism, 3 protein spots exclusively expressed in state of botulism, 3 protein spots were low-expressed in serum proteome under botulism. Fourteen proteins have been identified among 20 spots elected on two-dimensional electrophoresis gels. Crystal proteins family exclusively expressed in control group serum. Haptoglobin were low-expressed under botulism in serum protein components, however, serum amyloid A only expressed in serum sample under botulism in 24 h, which were verified by Western-blot. Identified proteins involved in energy metabolism, cellular stress response, transcription, body defense and cell proliferation. These findings represent the first report of BoNT-induced changes in serum proteome and histopathology, and reinforce the utility of applying proteomic tools to the study of system-wide biological processes in normal and botulism.

Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: implication for the interpretation of satellite bands.

PubMed

Hohenstein, Kurt; Griesmacher, Andrea; Weigel, Günter; Golderer, Georg; Ott, Helmut Werner

2011-06-01

Blue native electrophoresis (BNE) was applied to analyze the von Willebrand factor (vWF) multimers in their native state and to present a methodology to perform blue native electrophoresis on human plasma proteins, which has not been done before. The major difference between this method and the commonly used SDS-agarose gel electrophoresis is the lack of satellite bands in the high-resolution native gel. To further analyze this phenomenon, a second dimension was performed under denaturing conditions. Thereby, we obtained a pattern in which each protein sub-unit from the first dimension dissociates into three distinct sub-bands. These bands confirm the triplet structure, which consists of an intermediate band and two satellite bands. By introducing the second dimension, our novel method separates the triplet structure into a higher resolution than the commonly used SDS-agarose gel electrophoresis does. This helps considerably in the classification of ambiguous von Willebrand's disease subtypes. In addition, our method has the additional advantage of being able to resolve the triplet structure of platelet vWF multimers, which has not been identified previously through conventional SDS-agarose electrophoresis multimer analysis. This potential enables us to compare the triplet structure from platelet and plasmatic vWF, and may help to find out whether structural abnormalities concern the vWF molecule in the platelet itself, or whether they are due to the physiological processing of vWF shed into circulation. Owing to its resolution and sensitivity, this native separation technique offers a promising tool for the analysis and detection of von Willebrand disorder, and for the classification of von Willebrand's disease subtypes. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Influence of interface layer on optical properties of sub-20 nm-thick TiO2 films

NASA Astrophysics Data System (ADS)

Shi, Yue-Jie; Zhang, Rong-Jun; Li, Da-Hai; Zhan, Yi-Qiang; Lu, Hong-Liang; Jiang, An-Quan; Chen, Xin; Liu, Juan; Zheng, Yu-Xiang; Wang, Song-You; Chen, Liang-Yao

2018-02-01

The sub-20 nm ultrathin titanium dioxide (TiO2) films with tunable thickness were deposited on Si substrates by atomic layer deposition (ALD). The structural and optical properties were acquired by transmission electron microscopy, atomic force microscopy and spectroscopic ellipsometry. Afterwards, a constructive and effective method of analyzing interfaces by applying two different optical models consisting of air/TiO2/Ti x Si y O2/Si and air/effective TiO2 layer/Si, respectively, was proposed to investigate the influence of interface layer (IL) on the analysis of optical constants and the determination of band gap of TiO2 ultrathin films. It was found that two factors including optical constants and changing components of the nonstoichiometric IL could contribute to the extent of the influence. Furthermore, the investigated TiO2 ultrathin films of 600 ALD cycles were selected and then annealed at the temperature range of 400-900 °C by rapid thermal annealing. Thicker IL and phase transition cause the variation of optical properties of TiO2 films after annealing and a shorter electron relaxation time reveals the strengthened electron-electron and electron-phonon interactions in the TiO2 ultrathin films at high temperature. The as-obtained results in this paper will play a role in other studies of high dielectric constants materials grown on Si substrates and in the applications of next generation metal-oxide-semiconductor devices.

A Ga2O3 underlayer as an isomorphic template for ultrathin hematite films toward efficient photoelectrochemical water splitting.

PubMed

Hisatomi, Takashi; Brillet, Jérémie; Cornuz, Maurin; Le Formal, Florian; Tétreault, Nicolas; Sivula, Kevin; Grätzel, Michael

2012-01-01

Hematite photoanodes for photoelectrochemical (PEC) water splitting are often fabricated as extremely-thin films to minimize charge recombination because of the short diffusion lengths of photoexcited carriers. However, poor crystallinity caused by structural interaction with a substrate negates the potential of ultrathin hematite photoanodes. This study demonstrates that ultrathin Ga2O3 underlayers, which were deposited on conducting substrates prior to hematite layers by atomic layer deposition, served as an isomorphic (corundum-type) structural template for ultrathin hematite and improved the photocurrent onset of PEC water splitting by 0.2 V. The benefit from Ga2O3 underlayers was most pronounced when the thickness of the underlayer was approximately 2 nm. Thinner underlayers did not work effectively as a template presumably because of insufficient crystallinity of the underlayer, while thicker ones diminished the PEC performance of hematite because the underlayer prevented electron injection from hematite to a conductive substrate due to the large conduction band offset. The enhancement of PEC performance by a Ga2O3 underlayer was more significant for thinner hematite layers owing to greater margins for improving the crystallinity of ultrathin hematite. It was confirmed that a Ga2O3 underlayer was applicable to a rough conducting substrate loaded with Sb-doped SnO2 nanoparticles, improving the photocurrent by a factor of 1.4. Accordingly, a Ga2O3 underlayer could push forward the development of host-guest-type nanocomposites consisting of highly-rough substrates and extremely-thin hematite absorbers.

Simplification and improvement of protein detection in two-dimensional electrophoresis gels with SERVA HPE™ lightning red.

PubMed

Griebel, Anja; Obermaier, Christian; Westermeier, Reiner; Moche, Martin; Büttner, Knut

2013-07-01

A new fluorescent amino-reactive dye has been tested for both labelling proteins prior to electrophoretic separations and between the two steps of two-dimensional electrophoresis. A series of experiments showed, that the labelling of lysines with this dye is compatible with all standard additives used for sample preparation, including reducing substances and carrier ampholytes. Using this dye for pre-labelling considerably simplifies the electrophoresis and detection workflow and provides highly sensitive and quantitative visualisation of proteins.

Preparation of ultra-thin and high-quality WO{sub 3} compact layers and comparision of WO{sub 3} and TiO{sub 2} compact layer thickness in planar perovskite solar cells

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

Zhang, Jincheng; Shi, Chengwu, E-mail: shicw506@foxmail.com; Chen, Junjun

2016-06-15

In this paper, the ultra-thin and high-quality WO{sub 3} compact layers were successfully prepared by spin-coating-pyrolysis method using the tungsten isopropoxide solution in isopropanol. The influence of WO{sub 3} and TiO{sub 2} compact layer thickness on the photovoltaic performance of planar perovskite solar cells was systematically compared, and the interface charge transfer and recombination in planar perovskite solar cells with TiO{sub 2} compact layer was analyzed by electrochemical impedance spectroscopy. The results revealed that the optimum thickness of WO{sub 3} and TiO{sub 2} compact layer was 15 nm and 60 nm. The planar perovskite solar cell with 15 nm WO{submore » 3} compact layer gave a 9.69% average and 10.14% maximum photoelectric conversion efficiency, whereas the planar perovskite solar cell with 60 nm TiO{sub 2} compact layer achieved a 11.79% average and 12.64% maximum photoelectric conversion efficiency. - Graphical abstract: The planar perovskite solar cell with 15 nm WO{sub 3} compact layer gave a 9.69% average and 10.14% maximum photoelectric conversion efficiency, whereas the planar perovskite solar cell with 60 nm TiO{sub 2} compact layer achieved a 11.79% average and 12.64% maximum photoelectric conversion efficiency. Display Omitted - Highlights: • Preparation of ultra-thin and high-quality WO{sub 3} compact layers. • Perovskite solar cell with 15 nm-thick WO{sub 3} compact layer achieved PCE of 10.14%. • Perovskite solar cell with 60 nm-thick TiO{sub 2} compact layer achieved PCE of 12.64%.« less

Study of first electronic transition and hydrogen bonding state of ultra-thin water layer of nanometer thickness on an α-alumina surface by far-ultraviolet spectroscopy

NASA Astrophysics Data System (ADS)

Goto, Takeyoshi; Kinugasa, Tomoya

2018-05-01

The first electronic transition (A˜ ← X˜) and the hydrogen bonding state of an ultra-thin water layer of nanometer thickness between two α-alumina surfaces (0.5-20 nm) were studied using far-ultraviolet (FUV) spectroscopy in the wavelength range 140-180 nm. The ultra-thin water layer of nanometer thickness was prepared by squeezing a water droplet ( 1 μL) between a highly polished α-alumina prism and an α-alumina plate using a high pressure clamp ( 4.7 MPa), and the FUV spectra of the water layer at different thicknesses were measured using the attenuated total reflection method. As the water layer became thinner, the A˜ ← X˜ bands were gradually shifted to higher or lower energy relative to that of bulk water; at thicknesses smaller than 4 nm, these shifts were substantial (0.1-0.2 eV) in either case. The FUV spectra of the water layer with thickness < 4 nm indicate the formation of structured ice-like hydrogen bond (H-bond) layers for the higher energy shifts or the formation of slightly weaker H-bond layers as compared to those in the bulk liquid state for lower energy shifts. In either case, the H-bond structure of bulk liquid water is nearly lost at thicknesses below 4 nm, because of steric hydration forces between the α-alumina surfaces.

Chemical gating of epitaxial graphene through ultrathin oxide layers.

PubMed

Larciprete, Rosanna; Lacovig, Paolo; Orlando, Fabrizio; Dalmiglio, Matteo; Omiciuolo, Luca; Baraldi, Alessandro; Lizzit, Silvano

2015-08-07

We achieved a controllable chemical gating of epitaxial graphene grown on metal substrates by exploiting the electrostatic polarization of ultrathin SiO2 layers synthesized below it. Intercalated oxygen diffusing through the SiO2 layer modifies the metal-oxide work function and hole dopes graphene. The graphene/oxide/metal heterostructure behaves as a gated plane capacitor with the in situ grown SiO2 layer acting as a homogeneous dielectric spacer, whose high capacity allows the Fermi level of graphene to be shifted by a few hundreds of meV when the oxygen coverage at the metal substrate is of the order of 0.5 monolayers. The hole doping can be finely tuned by controlling the amount of interfacial oxygen, as well as by adjusting the thickness of the oxide layer. After complete thermal desorption of oxygen the intrinsic doping of SiO2 supported graphene is evaluated in the absence of contaminants and adventitious adsorbates. The demonstration that the charge state of graphene can be changed by chemically modifying the buried oxide/metal interface hints at the possibility of tuning the level and sign of doping by the use of other intercalants capable of diffusing through the ultrathin porous dielectric and reach the interface with the metal.

Investigation of embedded perovskite nanoparticles for enhanced capacitor permittivities.

PubMed

Krause, Andreas; Weber, Walter M; Pohl, Darius; Rellinghaus, Bernd; Verheijen, Marcel; Mikolajick, Thomas

2014-11-26

Growth experiments show significant differences in the crystallization of ultrathin CaTiO3 layers on polycrystalline Pt surfaces. While the deposition of ultrathin layers below crystallization temperature inhibits the full layer crystallization, local epitaxial growth of CaTiO3 crystals on top of specific oriented Pt crystals occurs. The result is a formation of crystals embedded in an amorphous matrix. An epitaxial alignment of the cubic CaTiO3 ⟨111⟩ direction on top of the underlying Pt {111} surface has been observed. A reduced forming energy is attributed to an interplay of surface energies at the {111} interface of both materials and CaTiO3 nanocrystallites facets. The preferential texturing of CaTiO3 layers on top of Pt has been used in the preparation of ultrathin metal-insulator-metal capacitors with 5-30 nm oxide thickness. The effective CaTiO3 permittivity in the capacitor stack increases to 55 compared to capacitors with amorphous layers and a permittivity of 28. The isolated CaTiO3 crystals exhibit a passivation of the CaTiO3 grain surfaces by the surrounding amorphous matrix, which keeps the capacitor leakage current at ideally low values comparable for those of amorphous thin film capacitors.

Interfacial engineering with ultrathin poly (9,9-di-n-octylfluorenyl-2,7-diyl) (PFO) layer for high efficient perovskite light-emitting diodes

NASA Astrophysics Data System (ADS)

Lin, Chunyan; Chen, Ping; Xiong, ZiYang; Liu, Debei; Wang, Gang; Meng, Yan; Song, Qunliang

2018-02-01

Organic-inorganic hybrid perovskites have attracted great attention in the field of lighting and display due to their very high color purity and low-cost solution-process. Researchers have done a lot of work in realizing high performance electroluminescent devices. However, the current efficiency (CE) of methyl-ammonium lead halide perovskite light-emitting diodes (PeLEDs) still needs to be improved. Herein, we demonstrate the enhanced performance of PeLEDs through introducing an ultrathin poly(9,9-di-n-octylfluorenyl-2,7-diyl) (PFO) buffer layer between poly(3,4-ethylendioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) and CH3NH3PbBr3 perovskite. Compared to the reference device without PFO, the optimal device luminous intensity, the maximum CE, and the maximum external quantum efficiency increases from 8139 cd m-2 to 30 150 cd m-2, from 7.20 cd A-1 (at 6.8 V) to 10.05 cd A-1 (at 6.6 V), and from 1.73% to 2.44%, respectively. The ultrathin PFO layer not only reduces the exciton quenching at the interface between the hole-transport layer and emission layer, but also passivates the shallow-trap ensure increasing hole injection, as well as increases the coverage of perovskite film.

In-Gel Stable-Isotope Labeling (ISIL): a strategy for mass spectrometry-based relative quantification.

PubMed

Asara, John M; Zhang, Xiang; Zheng, Bin; Christofk, Heather H; Wu, Ning; Cantley, Lewis C

2006-01-01

Most proteomics approaches for relative quantification of protein expression use a combination of stable-isotope labeling and mass spectrometry. Traditionally, researchers have used difference gel electrophoresis (DIGE) from stained 1D and 2D gels for relative quantification. While differences in protein staining intensity can often be visualized, abundant proteins can obscure less abundant proteins, and quantification of post-translational modifications is difficult. A method is presented for quantifying changes in the abundance of a specific protein or changes in specific modifications of a protein using In-gel Stable-Isotope Labeling (ISIL). Proteins extracted from any source (tissue, cell line, immunoprecipitate, etc.), treated under two experimental conditions, are resolved in separate lanes by gel electrophoresis. The regions of interest (visualized by staining) are reacted separately with light versus heavy isotope-labeled reagents, and the gel slices are then mixed and digested with proteases. The resulting peptides are then analyzed by LC-MS to determine relative abundance of light/heavy isotope pairs and analyzed by LC-MS/MS for identification of sequence and modifications. The strategy compares well with other relative quantification strategies, and in silico calculations reveal its effectiveness as a global relative quantification strategy. An advantage of ISIL is that visualization of gel differences can be used as a first quantification step followed by accurate and sensitive protein level stable-isotope labeling and mass spectrometry-based relative quantification.

Guine pig liver L-asparaginase. Separation, purification, and intracellular localisation of two distinct enzymatic activities.

PubMed

Rogez, J C; Plaquet, R; Biserte, G

1975-12-18

Two distinct L-asparaginase (EC 3.5.1.1) activities were detected in guinea pig liver: Asparaginase 1 and Asparaginase 2. Asparaginase 1 has been purified 272 fold from the crude homogenate; its molecular weight was evaluated by gel filtration to be about 150 000. The purified preparation was shown to be homogeneous by cellulose acetate strip and polyacrylamide disc-gel electrophoresis. Asparaginase 2 has been purified 63.5 fold from the crude homogenate. Its molecular weight was evaluated by gel filtration to be about 21 500. Cellulose acetate strip electrophoresis demonstrated two bands, one of which corresponded to Asparaginase 1 and the other to Asparaginase 2. Cellular fractionation in the ultracentrifuge, showed Asparaginase 1 to be present only in the cytosol fraction. Asparaginase 2 which was unstable at 105 000 X g seemed mostly localized in the mitochondria and secondarily in the cytoplasmic fraction.

Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis

NASA Technical Reports Server (NTRS)

Sutherland, Betsy M.; Georgakilas, Alexandros G.; Bennett, Paula V.; Laval, Jacques; Sutherland, John C.; Gewirtz, A. M. (Principal Investigator)

2003-01-01

Assessing DNA damage induction, repair and consequences of such damages requires measurement of specific DNA lesions by methods that are independent of biological responses to such lesions. Lesions affecting one DNA strand (altered bases, abasic sites, single strand breaks (SSB)) as well as damages affecting both strands (clustered damages, double strand breaks) can be quantified by direct measurement of DNA using gel electrophoresis, gel imaging and number average length analysis. Damage frequencies as low as a few sites per gigabase pair (10(9)bp) can be quantified by this approach in about 50ng of non-radioactive DNA, and single molecule methods may allow such measurements in DNA from single cells. This review presents the theoretical basis, biochemical requirements and practical aspects of this approach, and shows examples of their applications in identification and quantitation of complex clustered damages.

Purification and some properties of Fe protein of nitrogenase from. Anabaena cylindrica

NASA Astrophysics Data System (ADS)

Du, Daixian; Lin, Huimin; He, Zhenrong; Dai, Lingfen; Xin, Wusheng; Li, Shanghao

1990-12-01

The Fe protein of Anabaena cylindrica was first separated and purified by chromatography through DEAE-cellulose columns then by gel electrophoresis. The specific activity was up to 142.46 nmol C2H4/mg protein · min. It was homogeneous as shown by 1) a single band in the gel electrophorogram; 2) absence of Mo and tryptophan; 3) content of about 3.4 atoms of Fe per mole protein. The molecular weight of the Fe protein of A. cylindrica was about 61,000 daltons as estimated by SDS-gel electrophoresis and calculated from the amino acid composition. The residues of aspartate and glutamate were about 2.6 times that of arginine and lysine in the Fe protein. Crossing Fe protein of A. cylindrica with Mo-Fe protein of Azotobacter vinelandii gave positive result. The reciprocal crossing also showed activity.

A noda-like virus isolated from the sweetpotato pest spodoptera eridania (Cramer) (Lep.; noctuidae)

PubMed

Zeddam; Rodriguez; Ravallec; Lagnaoui

1999-11-01

A small isometric virus has been isolated from larvae of the sweetpotato pest Spodoptera eridania (Cramer) collected near Pariacoto, Ancash province, Peru. It is designated the Pariacoto virus (PaV). In addition to its high pathogenicity on its natural host Spodoptera eridania, PaV was found to replicate in Spodoptera ochrea (Hampson) larvae but not in Spodoptera frugiperda (Smith) larvae. The size of the viral particle was estimated to be about 30 nm in diameter. Polyacrylamide gel electrophoresis showed a protein of approximately 40.5 kDa. After agarose gel electrophoresis, the viral genome appeared to be bipartite RNA. Gel immunodiffusion tests showed no serological relationship between PaV and Nodamura virus, the type species for insect nodaviruses. Electron microscopy confirmed that viral replication occurs in the cytoplasm. These properties are similar to those of other members of family Nodaviridae, to which the virus is currently assigned. Copyright 1999 Academic Press.

Informatics and Statistics for Analyzing 2-D Gel Electrophoresis Images

PubMed Central

Dowsey, Andrew W.; Morris, Jeffrey S.; Gutstein, Howard B.; Yang, Guang-Zhong

2013-01-01

Despite recent progress in “shotgun” peptide separation by integrated liquid chromatography and mass spectrometry (LC/MS), proteome coverage and reproducibility are still limited with this approach and obtaining enough replicate runs for biomarker discovery is a challenge. For these reasons, recent research demonstrates that there is a continuing need for protein separation by two-dimensional gel electrophoresis (2-DE). However, with traditional 2-DE informatics, the digitized images are reduced to symbolic data through spot detection and quantification before proteins are compared for differential expression by spot matching. Recently, a more robust and automated paradigm has emerged where gels are directly aligned in the image domain before spots are detected across the whole image set as a whole. In this chapter, we describe the methodology for both approaches and discuss the pitfalls present when reasoning statistically about the differential protein expression discovered. PMID:20013375

Studies on the "Aerobic" Acetyl-Coenzyme A Synthetase of Saccharomyces Cerevisiae: Purification, Crystallization, and Physical Properties of the Enzyme

NASA Technical Reports Server (NTRS)

Satyanarayana, T.; Klein, Harold P.

1976-01-01

A procedure for the purification of a stable acetyl-coenzyme A synthetase (ACS) from aerobic cells of Saccharomyces cerevisiae is presented. The steps include differential centrifugation, solubilization of the bound enzyme from the crude mitochondrial fraction, ammonium sulfate fractionation, crystallization to constant specific activity from ammonium sulfate solutions followed by Bio-Gel A-1.5 m column chromatography. The resulting enzyme preparation is homogeneous as judged by chromatography on Bio-Gel columns, QAE-Sephadex A-50 anion exchange columns, analytical ultracentrifugal studies, and polyacrylamide gel electrophoresis. Sedimentation velocity runs revealed a single symmetric peak with an s(sub (20,w)) value of 10.6. The molecular weight of the native enzyme, as determined by gel filtration and analytical ultracentrifugation, is 250,000 +/- 500. In polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, the molecular weight of the single polypeptide chain is 83,000 +/- 500. The purified enzyme is inhibited by palmityl-coenzyme A with a Hill interaction coefficient, n, of 2.88. These studies indicate that the ACS of aerobic Saccharomyces cerevisiae is composed of three subunits of identical or nearly identical size.

Comparison of seven techniques for typing international epidemic strains of Clostridium difficile: restriction endonuclease analysis, pulsed-field gel electrophoresis, PCR-ribotyping, multilocus sequence typing, multilocus variable-number tandem-repeat analysis, amplified fragment length polymorphism, and surface layer protein A gene sequence typing.

PubMed

Killgore, George; Thompson, Angela; Johnson, Stuart; Brazier, Jon; Kuijper, Ed; Pepin, Jacques; Frost, Eric H; Savelkoul, Paul; Nicholson, Brad; van den Berg, Renate J; Kato, Haru; Sambol, Susan P; Zukowski, Walter; Woods, Christopher; Limbago, Brandi; Gerding, Dale N; McDonald, L Clifford

2008-02-01

Using 42 isolates contributed by laboratories in Canada, The Netherlands, the United Kingdom, and the United States, we compared the results of analyses done with seven Clostridium difficile typing techniques: multilocus variable-number tandem-repeat analysis (MLVA), amplified fragment length polymorphism (AFLP), surface layer protein A gene sequence typing (slpAST), PCR-ribotyping, restriction endonuclease analysis (REA), multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). We assessed the discriminating ability and typeability of each technique as well as the agreement among techniques in grouping isolates by allele profile A (AP-A) through AP-F, which are defined by toxinotype, the presence of the binary toxin gene, and deletion in the tcdC gene. We found that all isolates were typeable by all techniques and that discrimination index scores for the techniques tested ranged from 0.964 to 0.631 in the following order: MLVA, REA, PFGE, slpAST, PCR-ribotyping, MLST, and AFLP. All the techniques were able to distinguish the current epidemic strain of C. difficile (BI/027/NAP1) from other strains. All of the techniques showed multiple types for AP-A (toxinotype 0, binary toxin negative, and no tcdC gene deletion). REA, slpAST, MLST, and PCR-ribotyping all included AP-B (toxinotype III, binary toxin positive, and an 18-bp deletion in tcdC) in a single group that excluded other APs. PFGE, AFLP, and MLVA grouped two, one, and two different non-AP-B isolates, respectively, with their AP-B isolates. All techniques appear to be capable of detecting outbreak strains, but only REA and MLVA showed sufficient discrimination to distinguish strains from different outbreaks.

Insulator at the ultrathin limit: MgO on Ag(001).

PubMed

Schintke, S; Messerli, S; Pivetta, M; Patthey, F; Libioulle, L; Stengel, M; De Vita, A; Schneider, W D

2001-12-31

The electronic structure and morphology of ultrathin MgO films epitaxially grown on Ag(001) were investigated using low-temperature scanning tunneling spectroscopy and scanning tunneling microscopy. Layer-resolved differential conductance (dI/dU) measurements reveal that, even at a film thickness of three monolayers, a band gap of about 6 eV is formed corresponding to that of the MgO(001) single-crystal surface. This finding is confirmed by layer-resolved calculations of the local density of states based on density functional theory.

Optical bandgap of single- and multi-layered amorphous germanium ultra-thin films

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

Liu, Pei; Zaslavsky, Alexander; Longo, Paolo

2016-01-07

Accurate optical methods are required to determine the energy bandgap of amorphous semiconductors and elucidate the role of quantum confinement in nanometer-scale, ultra-thin absorbing layers. Here, we provide a critical comparison between well-established methods that are generally employed to determine the optical bandgap of thin-film amorphous semiconductors, starting from normal-incidence reflectance and transmittance measurements. First, we demonstrate that a more accurate estimate of the optical bandgap can be achieved by using a multiple-reflection interference model. We show that this model generates more reliable results compared to the widely accepted single-pass absorption method. Second, we compare two most representative methods (Taucmore » and Cody plots) that are extensively used to determine the optical bandgap of thin-film amorphous semiconductors starting from the extracted absorption coefficient. Analysis of the experimental absorption data acquired for ultra-thin amorphous germanium (a-Ge) layers demonstrates that the Cody model is able to provide a less ambiguous energy bandgap value. Finally, we apply our proposed method to experimentally determine the optical bandgap of a-Ge/SiO{sub 2} superlattices with single and multiple a-Ge layers down to 2 nm thickness.« less

Variable-Number Tandem Repeats That Are Useful in Genotyping Isolates of Salmonella enterica subsp. enterica Serovars Typhimurium and Newport▿

PubMed Central

Witonski, D. ; Stefanova, R.; Ranganathan, A.; Schutze, G. E.; Eisenach, K. D.; Cave, M. D.

2006-01-01

The genome of Salmonella enterica subsp. enterica serovar Typhimurium strain LT2 was analyzed for direct repeats, and 54 sequences containing variable-number tandem repeat loci were identified. Ten primer pairs that anneal upstream and downstream of each selected locus were designed and used to amplify PCR targets in isolates of S. enterica serovars Typhimurium and Newport. Four of the 10 loci did not show polymorphism in the length of products. Six loci were selected for analysis. Isolates of S. enterica serovars Typhimurium and Newport that were related to specific outbreaks and showed identical pulsed-field gel electrophoresis patterns were indistinguishable by the length of the six variable-number tandem repeats. Isolates that differed in their pulsed-field gel electrophoresis patterns showed polymorphism in variable-number tandem repeat profiles. Length of the products was confirmed by DNA sequence analysis. Only 2 of the 10 loci contained exact integers of the direct repeat. Eight loci contained partial copies. The partial copies were maintained at the ends of the variable-number tandem repeat loci in all isolates. In spite of having partial copies that were maintained in all isolates, the number of direct repeats at a locus was polymorphic. Six variable-number tandem repeat loci were useful in distinguishing isolates of S. enterica serovars Typhimurium and Newport that had different pulsed-field gel electrophoresis patterns and in identifying outbreak-associated cases that shared a common pulsed-field gel pattern. PMID:16943354

Accelerating monoenergetic protons from ultrathin foils by flat-top laser pulses in the directed-Coulomb-explosion regime

PubMed Central

Bulanov, S. S.; Brantov, A.; Bychenkov, V. Yu.; Chvykov, V.; Kalinchenko, G.; Matsuoka, T.; Rousseau, P.; Reed, S.; Yanovsky, V.; Litzenberg, D. W.; Krushelnick, K.; Maksimchuk, A.

2008-01-01

We consider the effect of laser beam shaping on proton acceleration in the interaction of a tightly focused pulse with ultrathin double-layer solid targets in the regime of directed Coulomb explosion. In this regime, the heavy ions of the front layer are forced by the laser to expand predominantly in the direction of the pulse propagation, forming a moving longitudinal charge separation electric field, thus increasing the effectiveness of acceleration of second-layer protons. The utilization of beam shaping, namely, the use of flat-top beams, leads to more efficient proton acceleration due to the increase of the longitudinal field. PMID:18850951

Ultrathin epitaxial barrier layer to avoid thermally induced phase transformation in oxide heterostructures

DOE PAGES

Baek, David J.; Lu, Di; Hikita, Yasuyuki; ...

2016-12-22

Incorporating oxides with radically different physical and chemical properties into heterostructures offers tantalizing possibilities to derive new functions and structures. Recently, we have fabricated freestanding 2D oxide membranes using the water-soluble perovskite Sr 3Al 2O 6 as a sacrificial buffer layer. Here, with atomic-resolution spectroscopic imaging, we observe that direct growth of oxide thin films on Sr 3Al 2O 6 can cause complete phase transformation of the buffer layer, rendering it water-insoluble. More importantly, we demonstrate that an ultrathin SrTiO 3 layer can be employed as an effective barrier to preserve Sr 3Al 2O 6 during subsequent growth, thus allowingmore » its integration in a wider range of oxide heterostructures.« less

Purification and investigation of some kinetic properties of glucose-6-phosphate dehydrogenase from parsley (Petroselinum hortense) leaves.

PubMed

Coban, T Abdül Kadir; Ciftçi, Mehmet; Küfrevioğlu, O Irfan

2002-05-01

In this study, glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP+ oxidoreductase, EC 1.1.1.49; G6PD) was purified from parsley (Petroselinum hortense) leaves, and analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps: preparation of homogenate, ammonium sulfate fractionation, and DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 8.79% and had a specific activity of 2.146 U (mg protein)(-1). The overall purification was about 58-fold. Temperature of +4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured according to the Beutler method, at 340 nm. In order to control the purification of enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acrylamide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for enzyme. The molecular weight was found to be 77.6 kDa by Sephadex G-150 gel filtration chromatography. A protein band corresponding to a molecular weight of 79.3 kDa was obtained on SDS-polyacrylamide gel electrophoresis. For the enzymes, the stable pH, optimum pH, and optimum temperature were found to be 6.0, 8.0, and 60 degrees C, respectively. Moreover, KM and Vmax values for NADP+ and G6-P at optimum pH and 25 degrees C were determined by means of Lineweaver-Burk graphs. Additionally, effects of streptomycin sulfate and tetracycline antibiotics were investigated for the enzyme activity of glucose-6-phosphate dehydrogenase in vitro.

Dimethylformamide interferes with Coomassie dye staining of proteins on blue native gel electrophoresis.

PubMed

Raghupathy, V; Oommen, Anna; Ramachandran, Anup

2014-06-15

Blue native gel electrophoresis (BN-PAGE) is used extensively for characterization of mitochondrial respiratory complexes and uses the binding of Coomassie brilliant blue G-250 to visualize proteins. Oxidative modification of sulfhydryl groups of such proteins can be evaluated by labeling with iodoacetamide conjugated to biotin (BIAM) and detected with streptavidin peroxidase on Western blots following BN-PAGE. However, dissolving BIAM in dimethylformamide, a recommended solvent, reduces Coomassie blue G staining to proteins during BN-PAGE. This interference is prevented by dissolving BIAM in dimethyl sulfoxide. Precautions in the use of the dye for protein staining subsequent to BIAM labeling are discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

Accommodating brightness and exposure levels in densitometry of stained polyacrylamide electrophoresis gels

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

Tan, Han Yen; Ng, Tuck Wah; Liew, Oi Wah

2010-03-20

Flatbed scanner densitometers can be operated under various illumination and recording exposure levels. In this work, we show that optical density measurement accuracy, sensitivity, and stability of stained polyacrylamide electrophoresis gel densitometry are crucially dependent on these two factors (brightness and exposure level), notwithstanding that the source is monochromatic, spatially uniform, and the measurements are made using an accurately calibrated step wedge in tandem. We further outline a method to accommodate the intensity deviations over a range of illumination and exposure levels in order to maintain sensitivity and repeatability in the computed optical densities. Comparisons were also made with resultsmore » from a commercial densitometer.« less

Imaging of Selenium by Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) in 2-D Electrophoresis Gels and Biological Tissues.

PubMed

Cruz, Elisa Castañeda Santa; Susanne Becker, J; Sabine Becker, J; Sussulini, Alessandra

2018-01-01

Selenium and selenoproteins are important components of living organisms that play a role in different biological processes. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is a powerful analytical technique that has been employed to obtain distribution maps of selenium in biological tissues in a direct manner, as well as in selenoproteins, previously separated by their molecular masses and isoelectric points using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). In this chapter, we present the protocols to perform LA-ICP-MS imaging experiments, allowing the distribution visualization and determination of selenium and/or selenoproteins in biological systems.

Analysis of bacterial populations in the environment using two-dimensional gel electrophoresis of genomic DNA and complementary DNA.

PubMed

Liu, Guo-Hua; Nakamura, Tatsuo; Amemiya, Takashi; Rajendran, Narasimmalu; Itoh, Kiminori

2011-01-01

Two-dimensional gel electrophoresis (2-DGE) mapping of genomic DNA and complementary DNA (cDNA) amplicons was attempted to analyze total and active bacterial populations within soil and activated sludge samples. Distinct differences in the number and species of bacterial populations and those that were metabolically active at the time of sampling were visually observed especially for the soil community. Statistical analyses and sequencing based on the 2-DGE data further revealed the relationships between total and active bacterial populations within each community. This high-resolution technique would be useful for obtaining a better understanding of bacterial population structures in the environment.

Changes induced by the Pepper mild mottle tobamovirus on the chloroplast proteome of Nicotiana benthamiana.

PubMed

Pineda, M; Sajnani, C; Barón, M

2010-01-01

We have analyzed the chloroplast proteome of Nicotiana benthamiana using two-dimensional gel electrophoresis and mass spectrometry followed by a database search. In order to improve the resolution of the two-dimensional electrophoresis gels, we have made separate maps for the low and the high pH range. At least 200 spots were detected. We identified 72 polypeptides, some being isoforms of different multiprotein families. In addition, changes in this chloroplast proteome induced by the infection with the Spanish strain of the Pepper mild mottle virus were investigated. Viral infection induced the down-regulation of several chloroplastidic proteins involved in both the photosynthetic electron-transport chain and the Benson-Calvin cycle.

Hybrid slab-microchannel gel electrophoresis system

DOEpatents

Balch, J.W.; Carrano, A.V.; Davidson, J.C.; Koo, J.C.

1998-05-05

A hybrid slab-microchannel gel electrophoresis system is described. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate. 4 figs.

Ultrathin Nitrogen-Doped Carbon Layer Uniformly Supported on Graphene Frameworks as Ultrahigh-Capacity Anode for Lithium-Ion Full Battery.

PubMed

Huang, Yanshan; Li, Ke; Yang, Guanhui; Aboud, Mohamed F Aly; Shakir, Imran; Xu, Yuxi

2018-03-01

The designable structure with 3D structure, ultrathin 2D nanosheets, and heteroatom doping are considered as highly promising routes to improve the electrochemical performance of carbon materials as anodes for lithium-ion batteries. However, it remains a significant challenge to efficiently integrate 3D interconnected porous frameworks with 2D tunable heteroatom-doped ultrathin carbon layers to further boost the performance. Herein, a novel nanostructure consisting of a uniform ultrathin N-doped carbon layer in situ coated on a 3D graphene framework (NC@GF) through solvothermal self-assembly/polymerization and pyrolysis is reported. The NC@GF with the nanosheets thickness of 4.0 nm and N content of 4.13 at% exhibits an ultrahigh reversible capacity of 2018 mA h g -1 at 0.5 A g -1 and an ultrafast charge-discharge feature with a remarkable capacity of 340 mA h g -1 at an ultrahigh current density of 40 A g -1 and a superlong cycle life with a capacity retention of 93% after 10 000 cycles at 40 A g -1 . More importantly, when coupled with LiFePO 4 cathode, the fabricated lithium-ion full cells also exhibit high capacity and excellent rate and cycling performances, highlighting the practicability of this NC@GF. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Modification of vital wheat gluten with phosphoric acid to produce high free solution capacity

USDA-ARS?s Scientific Manuscript database

Wheat gluten reacts with phosphoric acid in the presence of urea to produce natural superabsorbent gels. Fourier Transform Infra-red (FT-IR) spectroscopy and two-dimensional gel electrophoresis (2DE) reveal chemical changes from the reaction. Temperatures above 120°C and dry conditions create the op...

Application of a label-free, gel-free quantitative proteomics method for ecotoxicological studies of small fish species

EPA Science Inventory

Although two-dimensional electrophoresis (2D-GE) remains the basis for many ecotoxicoproteomic analyses, new, non gel-based methods are beginning to be applied to overcome throughput and coverage limitations of 2D-GE. The overall objective of our research was to apply a comprehe...

Fouling mechanisms of gel layer in a submerged membrane bioreactor.

PubMed

Hong, Huachang; Zhang, Meijia; He, Yiming; Chen, Jianrong; Lin, Hongjun

2014-08-01

The fouling mechanisms underlying gel layer formation and its filtration resistance in a submerged membrane bioreactor (MBR) were investigated. It was found that gel layer rather than cake layer was more easily formed when soluble microbial products content in sludge suspension was relatively high. Thermodynamic analyses showed that gel layer formation process should overcome a higher energy barrier as compared with cake layer formation process. However, when separation distance <2.3 nm, attractive interaction energy of gelling foulant-membrane combination was remarkably higher than that of sludge floc-membrane combination. The combined effects were responsible for gel layer formation. Filtration tests showed that specific filtration resistance (SFR) of gel layer was almost 100 times higher than that of cake layer. The unusually high SFR of gel layer could be ascribed to the gelling propensity and osmotic pressure mechanism. These findings shed significant light on fouling mechanisms of gel layer in MBRs. Copyright © 2014 Elsevier Ltd. All rights reserved.

High-Performance Ultrathin Active Chiral Metamaterials.

PubMed

Wu, Zilong; Chen, Xiaodong; Wang, Mingsong; Dong, Jianwen; Zheng, Yuebing

2018-05-22

Ultrathin active chiral metamaterials with dynamically tunable and responsive optical chirality enable new optical sensors, modulators, and switches. Herein, we develop ultrathin active chiral metamaterials of highly tunable chiroptical responses by inducing tunable near-field coupling in the metamaterials and exploit the metamaterials as ultrasensitive sensors to detect trace amounts of solvent impurities. To demonstrate the active chiral metamaterials mediated by tunable near-field coupling, we design moiré chiral metamaterials (MCMs) as model metamaterials, which consist of two layers of identical Au nanohole arrays stacked upon one another in moiré patterns with a dielectric spacer layer between the Au layers. Our simulations, analytical fittings, and experiments reveal that spacer-dependent near-field coupling exists in the MCMs, which significantly enhances the spectral shift and line shape change of the circular dichroism (CD) spectra of the MCMs. Furthermore, we use a silk fibroin thin film as the spacer layer in the MCM. With the solvent-controllable swelling of the silk fibroin thin films, we demonstrate actively tunable near-field coupling and chiroptical responses of the silk-MCMs. Impressively, we have achieved the spectral shift over a wavelength range that is more than one full width at half-maximum and the sign inversion of the CD spectra in a single ultrathin (1/5 of wavelength in thickness) MCM. Finally, we apply the silk-MCMs as ultrasensitive sensors to detect trace amounts of solvent impurities down to 200 ppm, corresponding to an ultrahigh sensitivity of >10 5 nm/refractive index unit (RIU) and a figure of merit of 10 5 /RIU.

Lipoprotein Agarose Gel Electrophoresis. Application in HDL-Cholesterol Methodology.

DTIC Science & Technology

1985-06-01

on determination of high-density lipo- protein cholesterol by precipitation with sodium phosphotungstate- magnesium. Clin Chem 25(4):560 (1979). 6...determinations of cholesterol levels in supernates obtained after addition of various amounts of precipitants , lipo- protein electrophoresis can help to...plotted against the corresponding volumes of sodium phosphotungstate-MgCl2 (NAPT) precipitant added. 10 ’ ’’ ’i - n

Understanding Electrophoresis through the Investigation of Size, Shape, and Charge of pH Indicators

ERIC Educational Resources Information Center

Brenner, Ryan K.; Hess, Kenneth R.; Morford, Jennifer L.

2015-01-01

A laboratory experiment was designed for upper-level students in a Chemical Analysis course to illustrate the theoretical and practical applications of 0.8% agarose gel electrophoresis and to reinforce an understanding of weak acids/bases using easy-to-visualize pH indicators. The careful choice of indicators included acid and base types with…

Factors affecting the separation performance of proteins in capillary electrophoresis.

PubMed

Zhu, Yueping; Li, Zhenqing; Wang, Ping; Shen, Lisong; Zhang, Dawei; Yamaguchi, Yoshinori

2018-04-15

Capillary electrophoresis (CE) is an effective tool for protein separation and analysis. Compared with capillary gel electrophoresis (CGE), non-gel sieving capillary electrophoresis (NGSCE) processes the superiority on operation, repeatability and automaticity. Herein, we investigated the effect of polymer molecular weight and concentration, electric field strength, and the effective length of the capillary on the separation performance of proteins, and find that (1) polymer with high molecular weight and concentration favors the separation of proteins, although concentrated polymer hinders its injection into the channel of the capillary due to its high viscosity. (2) The resolution between the adjacent proteins decreases with the increase of electric field strength. (3) When the effective length of the capillary is long, the separation performance improves at the cost of separation time. (4) 1.4% (w/v) hydroxyethyl cellulose (HEC), 100 V/cm voltage and 12 cm effective length offers the best separation for the proteins with molecular weight from 14,400 Da to 97,400 Da. Finally, we employed the optimal electrophoretic conditions to resolve Lysozyme, Ovalbumin, BSA and their mixtures, and found that they were baseline resolved within 15 min. Copyright © 2018 Elsevier B.V. All rights reserved.

Multiple-locus variable-number tandem-repeats analysis of Listeria monocytogenes using multicolour capillary electrophoresis and comparison with pulsed-field gel electrophoresis typing.

PubMed

Lindstedt, Bjørn-Arne; Tham, Wilhelm; Danielsson-Tham, Marie-Louise; Vardund, Traute; Helmersson, Seved; Kapperud, Georg

2008-02-01

The multiple-locus variable-number tandem-repeats analysis (MLVA) method for genotyping has proven to be a fast and reliable typing tool in several bacterial species. MLVA is in our laboratory the routine typing method for Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli O157. The gram-positive bacteria Listeria monocytogenes, while not isolated as frequent as S. Typhimurium and E. coli, causes severe illness with an overall mortality rate of 30%. Thus, it is important that any outbreak of this pathogen is detected early and a fast trace to the source can be performed. In view of this, we have used the information provided by two fully sequenced L. monocytogenes strains to develop a MLVA assay coupled with high-resolution capillary electrophoresis and compared it to pulsed-field gel electrophoresis (PFGE) in two sets of isolates, one Norwegian (79 isolates) and one Swedish (61 isolates) set. The MLVA assay could resolve all of the L. monocytogenes serotypes tested, and was slightly more discriminatory than PFGE for the Norwegian isolates (28 MLVA profiles and 24 PFGE profiles) and opposite for the Swedish isolates (42 MLVA profiles and 43 PFGE profiles).

Sulfate-reducing bacteria and their activities in cyanobacterial mats of Solar Lake (Sinai, Egypt)

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

Teske, A.; Ramsing, N.B.; Habicht, K.

1998-08-01

The sulfate-reducing bacteria within the surface layer of the hypersaline cyanobacterial mat of Solar Lake (Sinai, Egypt) were investigated with combined microbiological, molecular, and biogeochemical approaches. The diurnally oxic surface layer contained between 10{sup 6} and 10{sup 7} cultivable sulfate-reducing bacteria ml{sup {minus}1} day{sup {minus}1}, both in the same range as and sometimes higher than those in anaerobic deeper mat layers. In the oxic surface layer and in the mat layers below, filamentous sulfate-reducing Desulfonema bacteria were found in variable densities of 10{sup 4} and 10{sup 6} cells ml{sup {minus}1}. A Desulfonema-related, diurnally migrating bacterium was detected with PCR andmore » denaturing gradient gel electrophoresis within and below the oxic surface layer. Facultative aerobic respiration, filamentous morphology, motility, diurnal migration, and aggregate formation were the most conspicuous adaptations of Solar Lake sulfate-reducing bacteria to the mat matrix and to diurnal oxygen stress. A comparison of sulfate reduction rates within the mat and previously published photosynthesis rates showed that CO{sub 2} from sulfate reduction in the upper 5 mm accounted for 7 to 8% of the total photosynthetic CO{sub 2} demand of the mat.« less

On-surface synthesis: a promising strategy toward the encapsulation of air unstable ultra-thin 2D materials.

PubMed

Li, Qiang; Zhao, Yinghe; Guo, Jiyuan; Zhou, Qionghua; Chen, Qian; Wang, Jinlan

2018-02-22

2D black phosphorus (BP) and transition metal chalcogenides (TMCs) have beneficial electronic, optical, and physical properties at the few-layer limit. However, irreversible degradation of exfoliated or chemical vapor deposition-grown ultrathin BP and TMCs like GaSe via oxidation under ambient conditions limits their applications. Herein, the on-surface growth of an oxidation-resistant 2D thin film of a metal coordination polymer is demonstrated by multiscale simulations. We show that the preparation of such heterostructures can be conducted in solution, in which pristine BP and GaSe present better stability than in an air environment. Our calculations reveal that the interaction between the polymer layer and 2D materials is dominated by van der Waals forces; thus, the electronic properties of pristine BP and GaSe are well preserved. Meanwhile, the isolation from oxygen and water can be achieved by monolayer polymers, due to the nature of their close-packed layers. Our facile strategy for enhancing the environmental stability of ultrathin materials is expected to accelerate efforts to implement 2D materials in electronic and optoelectronic applications.

Ultra-thin distributed Bragg reflectors via stacked single-crystal silicon nanomembranes

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

Cho, Minkyu; Seo, Jung-Hun; Lee, Jaeseong

2015-05-04

In this paper, we report ultra-thin distributed Bragg reflectors (DBRs) via stacked single-crystal silicon (Si) nanomembranes (NMs). Mesh hole-free single-crystal Si NMs were released from a Si-on-insulator substrate and transferred to quartz and Si substrates. Thermal oxidation was applied to the transferred Si NM to form high-quality SiO{sub 2} and thus a Si/SiO{sub 2} pair with uniform and precisely controlled thicknesses. The Si/SiO{sub 2} layers, as smooth as epitaxial grown layers, minimize scattering loss at the interface and in between the layers. As a result, a reflection of 99.8% at the wavelength range from 1350 nm to 1650 nm can be measuredmore » from a 2.5-pair DBR on a quartz substrate and 3-pair DBR on a Si substrate with thickness of 0.87 μm and 1.14 μm, respectively. The high reflection, ultra-thin DBRs developed here, which can be applied to almost any devices and materials, holds potential for application in high performance optoelectronic devices and photonics applications.« less

Two-dimensional MoS2: A promising building block for biosensors.

PubMed

Gan, Xiaorong; Zhao, Huimin; Quan, Xie

2017-03-15

Recently, two-dimensional (2D) layered nanomaterials have trigged intensive interest due to the intriguing physicochemical properties that stem from a quantum size effect connected with their ultra-thin structure. In particular, 2D molybdenum disulfide (MoS 2 ), as an emerging class of stable inorganic graphene analogs with intrinsic finite bandgap, would possibly complement or even surpass graphene in electronics and optoelectronics fields. In this review, we first discuss the historical development of ultrathin 2D nanomaterials. Then, we are concerned with 2D MoS 2 including its structure-property relationships, synthesis methods, characterization for the layer thickness, and biosensor applications over the past five years. Thereinto, we are highlighting recent advances in 2D MoS 2 -based biosensors, especially emphasize the preparation of sensing elements, roles of 2D MoS 2 , and assay strategies. Finally, on the basis of the current achievements on 2D MoS 2 and other ultrathin layered nanomaterials, perspectives on the challenges and opportunities for the exploration of 2D MoS 2 -based biosensors are put forward. Copyright © 2016 Elsevier B.V. All rights reserved.

Ultraflexible organic amplifier with biocompatible gel electrodes.

PubMed

Sekitani, Tsuyoshi; Yokota, Tomoyuki; Kuribara, Kazunori; Kaltenbrunner, Martin; Fukushima, Takanori; Inoue, Yusuke; Sekino, Masaki; Isoyama, Takashi; Abe, Yusuke; Onodera, Hiroshi; Someya, Takao

2016-04-29

In vivo electronic monitoring systems are promising technology to obtain biosignals with high spatiotemporal resolution and sensitivity. Here we demonstrate the fabrication of a biocompatible highly conductive gel composite comprising multi-walled carbon nanotube-dispersed sheet with an aqueous hydrogel. This gel composite exhibits admittance of 100 mS cm(-2) and maintains high admittance even in a low-frequency range. On implantation into a living hypodermal tissue for 4 weeks, it showed a small foreign-body reaction compared with widely used metal electrodes. Capitalizing on the multi-functional gel composite, we fabricated an ultrathin and mechanically flexible organic active matrix amplifier on a 1.2-μm-thick polyethylene-naphthalate film to amplify (amplification factor: ∼200) weak biosignals. The composite was integrated to the amplifier to realize a direct lead epicardial electrocardiography that is easily spread over an uneven heart tissue.

STUDIES ON STRUCTURAL UNITS OF THE γ-GLOBULINS

PubMed Central

Edelman, G. M.; Poulik, M. D.

1961-01-01

When human and rabbit 7S γ-globulins were reduced in strong urea solutions by a number of procedures, their molecular weights fell to approximately ⅓ of the original values. Partial separation of the reduction products was achieved using chromatography and starch gel electrophoresis in urea solutions. One of the components of reduced human 7S γ-globulin was isolated by chromatography, identified by starch gel electrophoresis, and subjected to amino acid analyses. The amino acid composition of this component differed from that of the starting material and also from that of the remaining components. A reduced pathological macroglobulin dissociated to components with an average molecular weight of 41,000. Several reduced human myeloma proteins, when subjected to starch gel electrophoresis, yielded individual patterns that nevertheless had features in common with those of reduced normal γ-globulins. Reduction of normal and abnormal γ-globulins was accompanied by the appearance of titratable sulfhydryl groups. Chemical treatments other than reduction were used to determine the type of bond holding the subunits together. It was tentatively concluded that they were linked by disulfide bonds. An hypothesis is presented to relate the structural features of the various γ-globulins in terms of the multiplicity of polypeptide chains in these molecules. PMID:13725659

Assessment of the yeast species composition of cocoa bean fermentations in different cocoa-producing regions using denaturing gradient gel electrophoresis.

PubMed

Papalexandratou, Zoi; De Vuyst, Luc

2011-11-01

The yeast species composition of 12 cocoa bean fermentations carried out in Brazil, Ecuador, Ivory Coast and Malaysia was investigated culture-independently. Denaturing gradient gel electrophoresis of 26S rRNA gene fragments, obtained through polymerase chain reaction with universal eukaryotic primers, was carried out with two different commercial apparatus (the DCode and CBS systems). In general, this molecular method allowed a rapid monitoring of the yeast species prevailing during fermentation. Under similar and optimal denaturing gradient gel electrophoresis conditions, the CBS system allowed a better separated band pattern than the DCode system and an unambiguous detection of the prevailing species present in the fermentation samples. The most frequent yeast species were Hanseniaspora sp., followed by Pichia kudriavzevii and Saccharomyces cerevisiae, independent of the origin of the cocoa. This indicates a restricted yeast species composition of the cocoa bean fermentation process. Exceptionally, the Ivorian cocoa bean box fermentation samples showed a wider yeast species composition, with Hyphopichia burtonii and Meyerozyma caribbica among the main representatives. Yeasts were not detected in the samples when the temperature inside the fermenting cocoa pulp-bean mass reached values higher than 45 °C or under early acetic acid production conditions. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Microfluidic cell-phoresis enabling high-throughput analysis of red blood cell deformability and biophysical screening of antimalarial drugs.

PubMed

Santoso, Aline T; Deng, Xiaoyan; Lee, Jeong-Hyun; Matthews, Kerryn; Duffy, Simon P; Islamzada, Emel; McFaul, Sarah M; Myrand-Lapierre, Marie-Eve; Ma, Hongshen

2015-12-07

Changes in red blood cell (RBC) deformability are associated with the pathology of many diseases and could potentially be used to evaluate disease status and treatment efficacy. We developed a simple, sensitive, and multiplexed RBC deformability assay based on the spatial dispersion of single cells in structured microchannels. This mechanism is analogous to gel electrophoresis, but instead of transporting molecules through nano-structured material to measure their length, RBCs are transported through micro-structured material to measure their deformability. After transport, the spatial distribution of cells provides a readout similar to intensity bands in gel electrophoresis, enabling simultaneous measurement on multiple samples. We used this approach to study the biophysical signatures of falciparum malaria, for which we demonstrate label-free and calibration-free detection of ring-stage infection, as well as in vitro assessment of antimalarial drug efficacy. We show that clinical antimalarial drugs universally reduce the deformability of RBCs infected by Plasmodium falciparum and that recently discovered PfATP4 inhibitors, known to induce host-mediated parasite clearance, display a distinct biophysical signature. Our process captures key advantages from gel electrophoresis, including image-based readout and multiplexing, to provide a functional screen for new antimalarials and adjunctive agents.

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

PubMed

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

2007-08-01

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

Small acid soluble proteins for rapid spore identification.

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

Branda, Steven S.; Lane, Todd W.; VanderNoot, Victoria A.

2006-12-01

This one year LDRD addressed the problem of rapid characterization of bacterial spores such as those from the genus Bacillus, the group that contains pathogenic spores such as B. anthracis. In this effort we addressed the feasibility of using a proteomics based approach to spore characterization using a subset of conserved spore proteins known as the small acid soluble proteins or SASPs. We proposed developing techniques that built on our previous expertise in microseparations to rapidly characterize or identify spores. An alternative SASP extraction method was developed that was amenable to both the subsequent fluorescent labeling required for laser-induced fluorescencemore » detection and the low ionic strength requirements for isoelectric focusing. For the microseparations, both capillary isoelectric focusing and chip gel electrophoresis were employed. A variety of methods were evaluated to improve the molecular weight resolution for the SASPs, which are in a molecular weight range that is not well resolved by the current methods. Isoelectric focusing was optimized and employed to resolve the SASPs using UV absorbance detection. Proteomic signatures of native wild type Bacillus spores and clones genetically engineered to produce altered SASP patterns were assessed by slab gel electrophoresis, capillary isoelectric focusing with absorbance detection as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection.« less

Evaluation of agarose gel electrophoresis for characterization of silver nanoparticles in industrial products.

PubMed

Jimenez, Maria S; Luque-Alled, Jose M; Gomez, Teresa; Castillo, Juan R

2016-05-01

Agarose gel electrophoresis (AGE) has been used extensively for characterization of pure nanomaterials or mixtures of pure nanomaterials. We have evaluated the use of AGE for characterization of Ag nanoparticles (NPs) in an industrial product (described as strong antiseptic). Influence of different stabilizing agents (PEG, SDS, and sodium dodecylbenzenesulfonate), buffers (TBE and Tris Glycine), and functionalizing agents (mercaptosuccinic acid (TMA) and proteins) has been investigated for the characterization of AgNPs in the industrial product using different sizes-AgNPs standards. The use of 1% SDS, 0.1% TMA, and Tris Glycine in gel, electrophoresis buffer and loading buffer led to the different sizes-AgNPs standards moved according to their size/charge ratio (obtaining a linear relationship between apparent mobility and mean diameter). After using SDS and TMA, the behavior of the AgNPs in the industrial product (containing a casein matrix) was completely different, being not possible their size characterization. However we demonstrated that AGE with LA-ICP-MS detection is an alternative method to confirm the protein corona formation between the industrial product and two proteins (BSA and transferrin) maintaining NPs-protein binding (what is not possible using SDS-PAGE). © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Materials Science and Device Physics of 2-Dimensional Semiconductors

NASA Astrophysics Data System (ADS)

Fang, Hui

Materials and device innovations are the keys to future technology revolution. For MOSFET scaling in particular, semiconductors with ultra-thin thickness on insulator platform is currently of great interest, due to the potential of integrating excellent channel materials with the industrially mature Si processing. Meanwhile, ultra-thin thickness also induces strong quantum confinement which in turn affect most of the material properties of these 2-dimensional (2-D) semiconductors, providing unprecedented opportunities for emerging technologies. In this thesis, multiple novel 2-D material systems are explored. Chapter one introduces the present challenges faced by MOSFET scaling. Chapter two covers the integration of ultrathin III V membranes with Si. Free standing ultrathin III-V is studied to enable high performance III-V on Si MOSFETs with strain engineering and alloying. Chapter three studies the light absorption in 2-D membranes. Experimental results and theoretical analysis reveal that light absorption in the 2-D quantum membranes is quantized into a fundamental physical constant, where we call it the quantum unit of light absorption, irrelevant of most of the material dependent parameters. Chapter four starts to focus on another 2-D system, atomic thin layered chalcogenides. Single and few layered chalcogenides are first explored as channel materials, with focuses in engineering the contacts for high performance MOSFETs. Contact treatment by molecular doping methods reveals that many layered chalcogenides other than MoS2 exhibit good transport properties at single layer limit. Finally, Chapter five investigated 2-D van der Waals heterostructures built from different single layer chalcogenides. The investigation in a WSe2/MoS2 hetero-bilayer shows a large Stokes like shift between photoluminescence peak and lowest absorption peak, as well as strong photoluminescence intensity, consistent with spatially indirect transition in a type II band alignment in this van der Waals heterostructure. This result enables new family of semiconductor heterostructures having tunable optoelectronic properties with customized composite layers and highlights the ability to build van der Waals semiconductor heterostructure lasers/LEDs.

Inverted bulk-heterojunction organic solar cells with the transfer-printed anodes and low-temperature-processed ultrathin buffer layers

NASA Astrophysics Data System (ADS)

Itoh, Eiji; Sakai, Shota; Fukuda, Katsutoshi

2018-03-01

We studied the effects of a hole buffer layer [molybdenum oxide (MoO3) and natural copper oxide layer] and a low-temperature-processed electron buffer layer on the performance of inverted bulk-heterojunction organic solar cells in a device consisting of indium-tin oxide (ITO)/poly(ethylene imine) (PEI)/titanium oxide nanosheet (TiO-NS)/poly(3-hexylthiopnehe) (P3HT):phenyl-C61-butyric acid methylester (PCBM)/oxide/anode (Ag or Cu). The insertion of ultrathin TiO-NS (˜1 nm) and oxide hole buffer layers improved the open circuit voltage V OC, fill factor, and rectification properties owing to the effective hole blocking and electron transport properties of ultrathin TiO-NS, and to the enhanced work function difference between TiO-NS and the oxide hole buffer layer. The insertion of the TiO-NS contributed to the reduction in the potential barrier at the ITO/PEI/TiO-NS/active layer interface for electrons, and the insertion of the oxide hole buffer layer contributed to the reduction in the potential barrier for holes. The marked increase in the capacitance under positive biasing in the capacitance-voltage characteristics revealed that the combination of TiO-NS and MoO3 buffer layers contributes to the selective transport of electrons and holes, and blocks counter carriers at the active layer/oxide interface. The natural oxide layer of the copper electrode also acts as a hole buffer layer owing to the increase in the work function of the Cu surface in the inverted cells. The performance of the cell with evaporated MoO3 and Cu layers that were transfer-printed to the active layer was almost comparable to that of the cell with MoO3 and Ag layers directly evaporated onto the active layer. We also demonstrated comparable device performance in the cell with all-printed MoO3 and low-temperature-processed silver nanoparticles as an anode.

Sol-gel chemistry-based Ucon-coated columns for capillary electrophoresis.

PubMed

Hayes, J D; Malik, A

1997-07-18

A sol-gel chemistry-based novel approach for the preparation of a Ucon-coated fused-silica capillary column in capillary electrophoresis is presented. In this approach the sol-gel process is carried out inside 25 microm I.D. fused-silica capillaries. The sol solution contained appropriate quantities of an alkoxide-based sol-gel precursor, a polymeric coating material (Ucon), a crosslinking reagent, a surface derivatizing reagent, controlled amounts of water and a catalyst dissolved in a suitable solvent system. The coating procedure involves filling a capillary with the sol solution and allowing the sol-gel process to proceed for an optimum period. Hydrolysis of the alkoxide precursor and polycondensation of the hydrolyzed products with the surface silanol groups and the hydroxy-terminated Ucon molecules lead to the formation of a surface-bonded sol-gel coating on the inner walls of the capillary. The thickness of the coated film can be controlled by varying the reaction time, coating solution composition and experimental conditions. Commercial availability of high purity sol-gel precursors (e.g., TEOS 99.999%), the ease of coating, run-to-run and column-to-column reproducibility, and long column lifetimes make sol-gel coating chemistry very much suitable for being applied in analytical microseparations column technology. Test samples of basic proteins and nucleotides were used to evaluate the column performance. These results show that the sol-gel coating scheme has allowed for the generation of bio-compatible surfaces characterized by high separation efficiencies in CE. For different types of solutes, the sol-gel coated Ucon column consistently provided migration time R.S.D. values of the order of 0.5%.

Negative terahertz photoconductivity in 2D layered materials.

PubMed

Lu, Junpeng; Liu, Hongwei; Sun, Jing

2017-11-17

The remarkable qualities of 2D layered materials such as wide spectral coverage, high strength and great flexibility mean that ultrathin 2D layered materials have the potential to meet the criteria of next-generation optoelectronic devices. Photoconductivity is one of the critical parameters of materials applied to optoelectronics. In contrast to traditional semiconductors, specific ultrathin 2D layers present anomalous negative photoconductivity. This opens a new avenue for designing novel optoelectronic devices. It is important to have a deep understanding of the fundamentals of this anomalous response, in order to design and optimize such devices. In this review, we provide an overview of the observation of negative photoconductivity in 2D layered materials including graphene, topological insulators and transitional metal dichalcogenides. We also summarize recent reports on investigations into the fundamental mechanism using ultrafast terahertz (THz) spectroscopies. Finally, we conclude the review by discussing the existing challenges and proposing the possible prospects of this direction of research.

Electron Microscopy of Ultrathin Sections of Sporosarcina ureae

PubMed Central

Mazanec, K.; Kocur, M.; Martinec, T.

1965-01-01

Mazanec, K. (J. E. Purkyně University, Brno, Czechoslovakia), M. Kocur, and T. Martinec. Electron microscopy of ultrathin sections of Sporosarcina ureae. J. Bacteriol. 90:808–816. 1965.—Ultrathin sections of Sporosarcina ureae cells were studied by means of electron microscopy. The cell wall consists of several layers and is 340 A thick. The cytoplasm is of globular structure and includes ribosomelike structures, occasional mesosomes, and inclusions not precisely identifiable. The nuclear area has various shapes and is formed by filaments 10 to 20 A thick which proceed in various directions. Cell division occurs similarly to that of sarcinate. Both synchronic and asynchronic cell division was observed. The spores of S. ureae consist of an outer coat having several layers, a cortex, a spore wall, and cytoplasm. The results of the present investigation substantiate our previous suggestion that S. ureae should be transferred from the family Micrococcaceae to the family Bacillaceae. Images PMID:16562085

TOPICAL REVIEW: Ultra-thin film encapsulation processes for micro-electro-mechanical devices and systems

NASA Astrophysics Data System (ADS)

Stoldt, Conrad R.; Bright, Victor M.

2006-05-01

A range of physical properties can be achieved in micro-electro-mechanical systems (MEMS) through their encapsulation with solid-state, ultra-thin coatings. This paper reviews the application of single source chemical vapour deposition and atomic layer deposition (ALD) in the growth of submicron films on polycrystalline silicon microstructures for the improvement of microscale reliability and performance. In particular, microstructure encapsulation with silicon carbide, tungsten, alumina and alumina-zinc oxide alloy ultra-thin films is highlighted, and the mechanical, electrical, tribological and chemical impact of these overlayers is detailed. The potential use of solid-state, ultra-thin coatings in commercial microsystems is explored using radio frequency MEMS as a case study for the ALD alloy alumina-zinc oxide thin film.

Comparison of methods used to diagnose generalized inflammatory disease in manatees (Trichechus manatus latirostris)

USGS Publications Warehouse

Harr, K.E.; Harvey, J.W.; Bonde, R.K.; Murphy, D.; Lowe, Mark; Menchaca, M.; Haubold, E.M.; Francis-Floyd, R.

2006-01-01

Manatees (Trichechus manatus latirostris) are afflicted with inflammatory and infectious disease secondary to human interaction, such as boat strike and entanglement, as well as “cold stress syndrome” and pneumonia. White-blood-cell count and fever, primary indicators of systemic inflammation in most species, are insensitive in diagnosing inflammatory disease in manatees. Acute phase-response proteins, such as haptoglobin and serum amyloid A, have proven to be sensitive measures of inflammation/infection in domestic large animal species. This study assessed diagnosis of generalized inflammatory disease by different methods including total white-blood-cell count, albumin: globulin ratio, gel electrophoresis analysis, C-reactive protein, alpha1 acid glycoprotein, haptoglobin, fibrinogen, and serum amyloid A. Samples were collected from 71 apparently healthy and 27 diseased animals during diagnostic medical examination. Serum amyloid A, measured by ELISA, followed by albumin:globulin ratio, measured by plasma gel electrophoresis, were most sensitive in diagnosing inflammatory disease, with diagnostic sensitivity and specificity of approximately 90%. The reference interval for serum amyloid A is <10–50 μg/ml with an equivocal interval of 51–70 μg/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7–1.1. Albumin: globulin ratio, calculated using biochemical techniques, was not accurate due to overestimation of albumin by bromcresol green dye-binding methodology. Albumin:globulin ratio, measured by serum gel electrophoresis, has a low sensitivity of 15% due to the lack of fibrinogen in the sample. Haptoglobin, measured by hemoglobin titration, had a reference interval of 0.4–2.4 mg/ml, a diagnostic sensitivity of 60%, and a diagnostic specificity of 93%. The haptoglobin assay is significantly affected by hemolysis. Fibrinogen, measured by heat precipitation, has a reference interval of 100–400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%.

Comparison of methods used to diagnose generalized inflammatory disease in manatees (Trichechus manatus latirostris).

PubMed

Harr, Kendal; Harvey, John; Bonde, Robert; Murphy, David; Lowe, Mark; Menchaca, Maya; Haubold, Elsa; Francis-Floyd, Ruth

2006-06-01

Manatees (Trichechus manatus latirostris) are afflicted with inflammatory and infectious disease secondary to human interaction, such as boat strike and entanglement, as well as "cold stress syndrome" and pneumonia. White-blood-cell count and fever, primary indicators of systemic inflammation in most species, are insensitive in diagnosing inflammatory disease in manatees. Acute phase-response proteins, such as haptoglobin and serum amyloid A, have proven to be sensitive measures of inflammation/infection in domestic large animal species. This study assessed diagnosis of generalized inflammatory disease by different methods including total white-blood-cell count, albumin: globulin ratio, gel electrophoresis analysis, C-reactive protein, alpha, acid glycoprotein, haptoglobin, fibrinogen, and serum amyloid A. Samples were collected from 71 apparently healthy and 27 diseased animals during diagnostic medical examination. Serum amyloid A, measured by ELISA, followed by albumin:globulin ratio, measured by plasma gel electrophoresis, were most sensitive in diagnosing inflammatory disease, with diagnostic sensitivity and specificity of approximately 90%. The reference interval for serum amyloid A is <10-50 microg/ml with an equivocal interval of 51-70 microg/ml. The reference interval for albumin:globulin ratio by plasma gel electrophoresis is 0.7-1.1. Albumin: globulin ratio, calculated using biochemical techniques, was not accurate due to overestimation of albumin by bromcresol green dye-binding methodology. Albumin:globulin ratio, measured by serum gel electrophoresis, has a low sensitivity of 15% due to the lack of fibrinogen in the sample. Haptoglobin, measured by hemoglobin titration, had a reference interval of 0.4-2.4 mg/ml, a diagnostic sensitivity of 60%, and a diagnostic specificity of 93%. The haptoglobin assay is significantly affected by hemolysis. Fibrinogen, measured by heat precipitation, has a reference interval of 100-400 mg/dl, a diagnostic sensitivity of 40%, and a diagnostic specificity of 95%.

Nano-Photonic Structures for Light Trapping in Ultra-Thin Crystalline Silicon Solar Cells

PubMed Central

Pathi, Prathap; Peer, Akshit; Biswas, Rana

2017-01-01

Thick wafer-silicon is the dominant solar cell technology. It is of great interest to develop ultra-thin solar cells that can reduce materials usage, but still achieve acceptable performance and high solar absorption. Accordingly, we developed a highly absorbing ultra-thin crystalline Si based solar cell architecture using periodically patterned front and rear dielectric nanocone arrays which provide enhanced light trapping. The rear nanocones are embedded in a silver back reflector. In contrast to previous approaches, we utilize dielectric photonic crystals with a completely flat silicon absorber layer, providing expected high electronic quality and low carrier recombination. This architecture creates a dense mesh of wave-guided modes at near-infrared wavelengths in the absorber layer, generating enhanced absorption. For thin silicon (<2 μm) and 750 nm pitch arrays, scattering matrix simulations predict enhancements exceeding 90%. Absorption approaches the Lambertian limit at small thicknesses (<10 μm) and is slightly lower (by ~5%) at wafer-scale thicknesses. Parasitic losses are ~25% for ultra-thin (2 μm) silicon and just 1%–2% for thicker (>100 μm) cells. There is potential for 20 μm thick cells to provide 30 mA/cm2 photo-current and >20% efficiency. This architecture has great promise for ultra-thin silicon solar panels with reduced material utilization and enhanced light-trapping. PMID:28336851

Nano-photonic structures for light trapping in ultra-thin crystalline silicon solar cells

DOE PAGES

Pathi, Prathap; Peer, Akshit; Biswas, Rana

2017-01-13

Thick wafer-silicon is the dominant solar cell technology. It is of great interest to develop ultra-thin solar cells that can reduce materials usage, but still achieve acceptable performance and high solar absorption. Accordingly, we developed a highly absorbing ultra-thin crystalline Si based solar cell architecture using periodically patterned front and rear dielectric nanocone arrays which provide enhanced light trapping. The rear nanocones are embedded in a silver back reflector. In contrast to previous approaches, we utilize dielectric photonic crystals with a completely flat silicon absorber layer, providing expected high electronic quality and low carrier recombination. This architecture creates a densemore » mesh of wave-guided modes at near-infrared wavelengths in the absorber layer, generating enhanced absorption. For thin silicon (<2 μm) and 750 nm pitch arrays, scattering matrix simulations predict enhancements exceeding 90%. Absorption approaches the Lambertian limit at small thicknesses (<10 μm) and is slightly lower (by ~5%) at wafer-scale thicknesses. Parasitic losses are ~25% for ultra-thin (2 μm) silicon and just 1%–2% for thicker (>100 μm) cells. There is potential for 20 μm thick cells to provide 30 mA/cm2 photo-current and >20% efficiency. Furthermore, this architecture has great promise for ultra-thin silicon solar panels with reduced material utilization and enhanced light-trapping.« less

Nano-photonic structures for light trapping in ultra-thin crystalline silicon solar cells

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

Pathi, Prathap; Peer, Akshit; Biswas, Rana

Thick wafer-silicon is the dominant solar cell technology. It is of great interest to develop ultra-thin solar cells that can reduce materials usage, but still achieve acceptable performance and high solar absorption. Accordingly, we developed a highly absorbing ultra-thin crystalline Si based solar cell architecture using periodically patterned front and rear dielectric nanocone arrays which provide enhanced light trapping. The rear nanocones are embedded in a silver back reflector. In contrast to previous approaches, we utilize dielectric photonic crystals with a completely flat silicon absorber layer, providing expected high electronic quality and low carrier recombination. This architecture creates a densemore » mesh of wave-guided modes at near-infrared wavelengths in the absorber layer, generating enhanced absorption. For thin silicon (<2 μm) and 750 nm pitch arrays, scattering matrix simulations predict enhancements exceeding 90%. Absorption approaches the Lambertian limit at small thicknesses (<10 μm) and is slightly lower (by ~5%) at wafer-scale thicknesses. Parasitic losses are ~25% for ultra-thin (2 μm) silicon and just 1%–2% for thicker (>100 μm) cells. There is potential for 20 μm thick cells to provide 30 mA/cm2 photo-current and >20% efficiency. Furthermore, this architecture has great promise for ultra-thin silicon solar panels with reduced material utilization and enhanced light-trapping.« less

Ultrathin layered double hydroxide nanosheets with Ni(III) active species obtained by exfoliation for highly efficient ethanol electrooxidation.

PubMed

Xu, Liang; Wang, Zhe; Chen, Xu; Qu, Zongkai; Li, Feng; Yang, Wensheng

2018-01-10

The development of non-precious metal electrocatalysts for renewable energy conversion and storage is compelling but greatly challenging due to low activity of the existing catalysts. Herein, the ultrathin NiAl-layered double hydroxide nanosheets (NiAl-LDH-NSs) are prepared by simple liquid-exfoliation of bulk NiAl-LDHs and first used as ethanol electrooxidation catalysts. The ultrathin two-dimensional (2D) structure ensures that the LDH nanosheets expose a greater number of active sites. More importantly, much Ni(III) active species (NiOOH) in the ultrathin nanosheets are formed by the exfoliation process, which play an authentic catalytic role in the ethanol oxidation reaction (EOR). The presence of NiOOH remarkably improves the reactivity and electrical conductivity of LDH nanosheets. These synergistic effects lead to strikingly more than 30 times enhanced EOR activity of NiAl-LDH-NSs compared to bulk NiAl-LDHs. The obtained electrocatalytic activity is also much better than those of most Ni- and LDH-based EOR catalysts reported to date. In addition, the ultrathin NiAl-LDH-NS electrocatalyst also exhibits good long-term stability (maintain 81.8% of the original value after 10000 s). This study not only provides a highly competitive EOR catalyst, but also opens new avenues toward the design of highly efficient electrode materials that have various potential applications in supercapacitor, Ni-MH battery and other electrocatalytic systems.

Nano-Photonic Structures for Light Trapping in Ultra-Thin Crystalline Silicon Solar Cells.

PubMed

Pathi, Prathap; Peer, Akshit; Biswas, Rana

2017-01-13

Thick wafer-silicon is the dominant solar cell technology. It is of great interest to develop ultra-thin solar cells that can reduce materials usage, but still achieve acceptable performance and high solar absorption. Accordingly, we developed a highly absorbing ultra-thin crystalline Si based solar cell architecture using periodically patterned front and rear dielectric nanocone arrays which provide enhanced light trapping. The rear nanocones are embedded in a silver back reflector. In contrast to previous approaches, we utilize dielectric photonic crystals with a completely flat silicon absorber layer, providing expected high electronic quality and low carrier recombination. This architecture creates a dense mesh of wave-guided modes at near-infrared wavelengths in the absorber layer, generating enhanced absorption. For thin silicon (<2 μm) and 750 nm pitch arrays, scattering matrix simulations predict enhancements exceeding 90%. Absorption approaches the Lambertian limit at small thicknesses (<10 μm) and is slightly lower (by ~5%) at wafer-scale thicknesses. Parasitic losses are ~25% for ultra-thin (2 μm) silicon and just 1%-2% for thicker (>100 μm) cells. There is potential for 20 μm thick cells to provide 30 mA/cm² photo-current and >20% efficiency. This architecture has great promise for ultra-thin silicon solar panels with reduced material utilization and enhanced light-trapping.

New Strategies and Methods to Study Interactions between Tobacco Mosaic Virus Coat Protein and Its Inhibitors

PubMed Central

Li, Xiangyang; Chen, Zhuo; Jin, Linhong; Hu, Deyu; Yang, Song

2016-01-01

Studies of the targets of anti-viral compounds are hot topics in the field of pesticide research. Various efficient anti-TMV (Tobacco Mosaic Virus) compounds, such as Ningnanmycin (NNM), Antofine (ATF), Dufulin (DFL) and Bingqingxiao (BQX) are available. However, the mechanisms of the action of these compounds on targets remain unclear. To further study the mechanism of the action of the anti-TMV inhibitors, the TMV coat protein (TMV CP) was expressed and self-assembled into four-layer aggregate disks in vitro, which could be reassembled into infectious virus particles with TMV RNA. The interactions between the anti-TMV compounds and the TMV CP disk were analyzed by size exclusion chromatography, isothermal titration calorimetry and native-polyacrylamide gel electrophoresis methods. The results revealed that assembly of the four-layer aggregate disk was inhibited by NNM; it changed the four-layer aggregate disk into trimers, and affected the regular assembly of TMV CP and TMV RNA. The four-layer aggregate disk of TMV CP was little inhibited by ATF, DFL and BQX. Our results provide original data, as well as new strategies and methods, for research on the mechanism of action of anti-viral drugs. PMID:26927077

A surface-mediated siRNA delivery system developed with chitosan/hyaluronic acid-siRNA multilayer films through layer-by-layer self-assembly

NASA Astrophysics Data System (ADS)

Wu, Lijuan; Wu, Changlin; Liu, Guangwan; Liao, Nannan; Zhao, Fang; Yang, Xuxia; Qu, Hongyuan; Peng, Bo; Chen, Li; Yang, Guang

2016-12-01

siRNA delivery remains highly challenging because of its hydrophilic and anionic nature and its sensitivity to nuclease degradation. Effective siRNA loading and improved transfection efficiency into cells represents a key problem. In our study, we prepared Chitosan/Hyaluronic acid-siRNA multilayer films through layer-by-layer self-assembly, in which siRNAs can be effectively loaded and protected. The construction process was characterized by FTIR, 13C NMR (CP/MAS), UV-vis spectroscopy, and atomic force microscopy (AFM). We presented the controlled-release performance of the films during incubation in 1 M NaCl solution for several days through UV-vis spectroscopy and polyacrylamide gel electrophoresis (PAGE). Additionally, we verified the stability and integrity of the siRNA loaded on multilayer films. Finally, the biological efficacy of the siRNA delivery system was evaluated via cells adhesion and gene silencing analyses in eGFP-HEK 293T cells. This new type of surface-mediated non-viral multilayer films may have considerable potential in the localized and controlled-release delivery of siRNA in mucosal tissues, and tissue engineering application.

Protamine ratio and the level of histone retention in sperm selected from a density gradient preparation.

PubMed

Hammoud, S; Liu, L; Carrell, D T

2009-04-01

Fertile males express two forms of sperm nuclear proteins, protamine 1 (P1) and protamine 2 (P2), in roughly equal quantities, whereas some infertile men have been shown to have a reduction in protamine content and an increase in the level of histones retained in mature sperm. In this study, we assessed histone and protamine levels in spermatozoa isolated from different layers of a density gradient centrifugation column to evaluate the nuclear protein content of the sperm population selected. Protamine levels were measured using acid gel electrophoresis and immunofluorescence, and the percentage of cells retaining histones was evaluated using aniline staining and immunofluorescence. Our data suggests that there is an inverse correlation between P1/P2 ratio and the level of histone expression in the different layers of the density gradient. Paradoxically, the 90% layer had a lower P1/P2 ratio, which corresponded with an increase in histone expression. It is concluded that although the sperm population selected in the 90% layer of the density gradient columns had a lower P1/P2 ratio, it was yet similar to the P1/P2 ratio observed in previously screened fertile donors.

Agarose and Polyacrylamide Gel Electrophoresis Methods for Molecular Mass Analysis of 5–500 kDa Hyaluronan

PubMed Central

Bhilocha, Shardul; Amin, Ripal; Pandya, Monika; Yuan, Han; Tank, Mihir; LoBello, Jaclyn; Shytuhina, Anastasia; Wang, Wenlan; Wisniewski, Hans-Georg; de la Motte, Carol; Cowman, Mary K.

2011-01-01

Agarose and polyacrylamide gel electrophoresis systems for the molecular mass-dependent separation of hyaluronan (HA) in the size range of approximately 5–500 kDa have been investigated. For agarose-based systems, the suitability of different agarose types, agarose concentrations, and buffers systems were determined. Using chemoenzymatically synthesized HA standards of low polydispersity, the molecular mass range was determined for each gel composition, over which the relationship between HA mobility and logarithm of the molecular mass was linear. Excellent linear calibration was obtained for HA molecular mass as low as approximately 9 kDa in agarose gels. For higher resolution separation, and for extension to molecular masses as low as approximately 5 kDa, gradient polyacrylamide gels were superior. Densitometric scanning of stained gels allowed analysis of the range of molecular masses present in a sample, and calculation of weight-average and number-average values. The methods were validated for polydisperse HA samples with viscosity-average molecular masses of 112, 59, 37, and 22 kDa, at sample loads of 0.5 µg (for polyacrylamide) to 2.5 µg (for agarose). Use of the methods for electrophoretic mobility shift assays was demonstrated for binding of the HA-binding region of aggrecan (recombinant human aggrecan G1-IGD-G2 domains) to a 150 kDa HA standard. PMID:21684248

Detection of sequence variation in parasite ribosomal DNA by electrophoresis in agarose gels supplemented with a DNA-intercalating agent.

PubMed

Zhu, X Q; Chilton, N B; Gasser, R B

1998-05-01

This study evaluated the use of a commercially available DNA intercalating agent (Resolver Gold) in agarose gels for the direct detection of sequence variation in ribosomal DNA (rDNA). This agent binds preferentially to AT sequence motifs in DNA. Regions of nuclear rDNA, known to provide genetic markers for the identification of species of parasitic ascarid nematodes (order Ascaridida), were amplified by polymerase chain reaction (PCR) and subjected to electrophoresis in standard agarose gels versus gels supplemented with Resolver Gold. Individual taxa examined could not be distinguished reliably based on the size of their amplicons in standard agarose gels, whereas they could be readily delineated based on mobility using Resolver Gold-supplemented gels. The latter was achieved because of differences (approximately 0.1-8.2%) in the AT content of the fragments among different taxa, which were associated with significant interspecific differences (approximately 11-39%) in the rDNA sequences employed. There was a tendency for fragments with higher AT content to migrate slower in supplemented agarose gels compared with those of lower AT content. The results indicate the usefulness of this electrophoretic approach to rapidly screen for sequence variability within or among PCR-amplified rDNA fragments of similar sizes but differing AT contents. Although evaluated on rDNA of parasites, the approach has potential to be applied to a range of genes of different groups of infectious organisms.

Urine Collected From Diapers Can Be Used for 2-Dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) in Infants and Young Children

PubMed Central

Kennedy, Mary Jayne; Griffin, Angela; Su, Ruifeng; Merchant, Michael; Klein, Jon

2011-01-01

Urinary proteomic profiling has potential to identify candidate biomarkers of renal injury in infants provided an adequate urine sample can be obtained. Although diapers are used to obtain urine for clinical evaluation, their use for proteomic analysis has not been investigated. We therefore performed feasibility studies on the use of diaper-extracted urine for 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Pediatric waste urine (2–20 mL) was applied to gel-containing, non-gel and cotton-gauze diapers and then mechanically expressed. Urine volume and total protein were measured pre- and post-extraction. Proteins were separated via 2D-PAGE following application of urine (20–40 mL) to each matrix. 2D-PAGE was also performed on clinical specimens collected using each diaper type. Differences in the adsorption and retention of urine volume and protein were noted between matrices. Non-gel and cotton-gauze diapers provided the best protein/volume recovery and the lowest interference with the Bradford assay. 2D-PAGE was also successfully completed using urine samples from both cotton fiber matrices. Conversely, samples from low-gel diapers demonstrated poor protein separation and reproducibility. Diapers containing cotton-fiber matrices appear adequate for 2D-PAGE. Qualitative and quantitative analyses of resolved proteins using replicate, high resolution gels will be required, however, before diaper-extracted urine can be applied in proteomic profiling. PMID:21137001

GaAs shallow-homojunction solar cells

NASA Technical Reports Server (NTRS)

Fan, J. C. C.

1981-01-01

The feasibility of fabricating space resistant, high efficiency, light weight, low cost GaAs shallow homojunction solar cells for space application is investigated. The material preparation of ultrathin GaAs single crystal layers, and the fabrication of efficient GaAs solar cells on bulk GaAs substrates are discussed. Considerable progress was made in both areas, and conversion efficiency about 16% AMO was obtained using anodic oxide as a single layer antireflection coating. A computer design shows that even better cells can be obtained with double layer antireflection coating. Ultrathin, high efficiency solar cells were obtained from GaAs films prepared by the CLEFT process, with conversion efficiency as high as 17% at AMI from a 10 micrometers thick GaAs film. A organometallic CVD was designed and constructed.

Disorder-controlled superconductivity at YBa2Cu3O7/SrTiO3 interfaces

NASA Astrophysics Data System (ADS)

Garcia-Barriocanal, J.; Perez-Muñoz, A. M.; Sefrioui, Z.; Arias, D.; Varela, M.; Leon, C.; Pennycook, S. J.; Santamaria, J.

2013-06-01

We examine the effect of interface disorder in suppressing superconductivity in coherently grown ultrathin YBa2Cu3O7 (YBCO) layers on SrTiO3 (STO) in YBCO/STO superlattices. The termination plane of the STO is TiO2 and the CuO chains are missing at the interface. Disorder (steps) at the STO interface cause alterations of the stacking sequence of the intracell YBCO atomic layers. Stacking faults give rise to antiphase boundaries which break the continuity of the CuO2 planes and depress superconductivity. We show that superconductivity is directly controlled by interface disorder outlining the importance of pair breaking and localization by disorder in ultrathin layers.

Tunneling interferometry and measurement of the thickness of ultrathin metallic Pb(111) films

NASA Astrophysics Data System (ADS)

Ustavshchikov, S. S.; Putilov, A. V.; Aladyshkin, A. Yu.

2017-10-01

Spectra of the differential tunneling conductivity for ultrathin lead films grown on Si(111) 7 × 7 single crystals with a thickness of 9 to 50 ML have been studied by low-temperature scanning tunneling microscopy and spectroscopy. The presence of local maxima of the tunneling conductivity is characteristic of such systems. The energies of maxima of the differential conductivity are determined by the spectrum of quantum-confined states of electrons in a metallic layer and, consequently, the local thickness of the layer. It has been shown that features of the microstructure of substrates, such as steps of monatomic height, structural defects, and inclusions of other materials covered with a lead layer, can be visualized by bias-modulation scanning tunneling spectroscopy.

Heterogeneity in ultrathin films simulated by Monte Carlo method

NASA Astrophysics Data System (ADS)

Sun, Jiebing; Hannon, James B.; Kellogg, Gary L.; Pohl, Karsten

2007-03-01

The 3D composition profile of ultra-thin Pd films on Cu(001) has been experimentally determined using low energy electron microscopy (LEEM).^[1] Quantitative measurements of the alloy concentration profile near steps show that the Pd distribution in the 3^rd layer is heterogeneous due to step overgrowth during Pd deposition. Interestingly, the Pd distribution in the 2^nd layer is also heterogeneous, and appears to be correlated with the distribution in the 1^st layer. We describe Monte Carlo simulations that show that correlation is due to Cu-Pd attraction, and that the 2^nd layer Pd is, in fact, laterally equilibrated. By comparing measured and simulated concentration profiles, we can estimate this attraction within a simple bond counting model. [1] J. B. Hannon, J. Sun, K. Pohl, G. L. Kellogg, Phys. Rev. Lett. 96, 246103 (2006)

Protein composition of wheat gluten polymer fractions determined by quantitative two-dimensional gel electrophoresis and tandem mass spectrometry

USDA-ARS?s Scientific Manuscript database

Flour proteins from the US bread wheat Butte 86 were extracted in 0.5% SDS using a two-step procedure with and without sonication and further separated by size exclusion chromatography into monomeric and polymeric fractions. Proteins in each fraction were analyzed by quantitative two-dimensional gel...

Modeling the Dynamics of Gel Electrophorresis in the High School Classroom

ERIC Educational Resources Information Center

Saucedo, Skyler R.

2013-01-01

Gel electrophoresis, used by geneticists and forensic experts alike, is an immensely popular technique that utilizes an electric field to separate molecules and proteins by size and charge. At the microscopic level, a dye or complex protein like DNA is passed through agarose, a gelatinous three-dimensional matrix of pores and nano-sized tunnels.…

Management of light absorption in extraordinary optical transmission based ultra-thin-film tandem solar cells

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

Mashooq, Kishwar; Talukder, Muhammad Anisuzzaman, E-mail: anis@eee.buet.ac.bd

2016-05-21

Although ultra-thin-film solar cells can be attractive in reducing the cost, they suffer from low absorption as the thickness of the active layer is usually much smaller than the wavelength of incident light. Different nano-photonic techniques, including plasmonic structures, are being explored to increase the light absorption in ultra-thin-film solar cells. More than one layer of active materials with different energy bandgaps can be used in tandem to increase the light absorption as well. However, due to different amount of light absorption in different active layers, photo-generated currents in different active layers will not be the same. The current mismatchmore » between the tandem layers makes them ineffective in increasing the efficiency. In this work, we investigate the light absorption properties of tandem solar cells with two ultra-thin active layers working as two subcells and a metal layer with periodically perforated holes in-between the two subcells. While the metal layer helps to overcome the current mismatch, the periodic holes increase the absorption of incident light by helping extraordinary optical transmission of the incident light from the top to the bottom subcell, and by coupling the incident light to plasmonic and photonic modes within ultra-thin active layers. We extensively study the effects of the geometry of holes in the intermediate metal layer on the light absorption properties of tandem solar cells with ultra-thin active layers. We also study how different metals in the intermediate layer affect the light absorption; how the geometry of holes in the intermediate layer affects the absorption when the active layer materials are changed; and how the intermediate metal layer affects the collection of photo-generated electron-hole pairs at the terminals. We find that in a solar cell with 6,6-phenyl C61-butyric acid methyl ester top subcell and copper indium gallium selenide bottom subcell, if the periodic holes in the metal layer are square or polygon, total absorption remains approximately the same. However, the total absorption suffers significantly if the holes are triangle. The transmission spectra of incident light into the bottom subcell, and hence the absorption, change significantly for square and circle holes if the active materials change to cadmium selenide (CdSe) and cadmium telluride (CdTe) in the top and bottom subcells, respectively. Although the intermediate metal layer may induce electron-hole pair recombination due to surface defects, the short-circuit current density of an ultra-thin plasmonic solar cell with an intermediate metal layer with two-dimensional hole array is >9% of that of a structure without the intermediate metal layer.« less

Application of 2-dimensional difference gel electrophoresis (2D-DIGE) to the study of thrombin-activated human platelet secretome.

PubMed

Della Corte, Anna; Maugeri, Norma; Pampuch, Agnieszka; Cerletti, Chiara; de Gaetano, Giovanni; Rotilio, Domenico

2008-02-01

Thrombin is an agonist inducing platelet activation. We combined two-dimensional difference gel electrophoresis (2D-DIGE) and mass spectrometry (MALDI-TOF MS) to analyse differentially expressed proteins secreted from thrombin-stimulated platelets. Human washed platelets, from healthy volunteers, were stimulated with thrombin 0.5 U/ml at 37 degrees C without stirring and the secreted proteins were resolved by 2D-DIGE. By image analysis, 1094 spots were detected in the 2D gel. The spots whose mean intensity showed at least a five-fold change intensity increase or decrease in the thrombin-activated platelet gel in comparison with unstimulated control were digested by trypsin and subjected to MALDI-TOF MS analysis. Peptides from mass spectra of in-gel digest samples were matched against available databases, using the Mascot search engine (Matrix Science) for peptide mass fingerprint. In the activated platelet secretome, transferrin, glutathione-transferase, WD repeat protein, ER-60, thrombospondin-1 precursor and thrombospondin were the most abundant. Also lamin A, a nuclear protein, not previously identified in platelets, appeared to be released. The novel strategy to combine 2D-DIGE with MALDI-TOF MS is a useful approach for a quantitative analysis of the effect of thrombin on the secretome profile of human platelets.

Development of an open source laboratory information management system for 2-D gel electrophoresis-based proteomics workflow

PubMed Central

Morisawa, Hiraku; Hirota, Mikako; Toda, Tosifusa

2006-01-01

Background In the post-genome era, most research scientists working in the field of proteomics are confronted with difficulties in management of large volumes of data, which they are required to keep in formats suitable for subsequent data mining. Therefore, a well-developed open source laboratory information management system (LIMS) should be available for their proteomics research studies. Results We developed an open source LIMS appropriately customized for 2-D gel electrophoresis-based proteomics workflow. The main features of its design are compactness, flexibility and connectivity to public databases. It supports the handling of data imported from mass spectrometry software and 2-D gel image analysis software. The LIMS is equipped with the same input interface for 2-D gel information as a clickable map on public 2DPAGE databases. The LIMS allows researchers to follow their own experimental procedures by reviewing the illustrations of 2-D gel maps and well layouts on the digestion plates and MS sample plates. Conclusion Our new open source LIMS is now available as a basic model for proteome informatics, and is accessible for further improvement. We hope that many research scientists working in the field of proteomics will evaluate our LIMS and suggest ways in which it can be improved. PMID:17018156

Few-layered CoHPO4 · 3H2O ultrathin nanosheets for high performance of electrode materials for supercapacitors.

PubMed

Pang, Huan; Wang, Shaomei; Shao, Weifang; Zhao, Shanshan; Yan, Bo; Li, Xinran; Li, Sujuan; Chen, Jing; Du, Weimin

2013-07-07

Ultrathin cobalt phosphate (CoHPO4 · 3H2O) nanosheets are successfully synthesized by a one pot hydrothermal method. Novel CoHPO4 · 3H2O ultrathin nanosheets are assembled for constructing the electrodes of supercapacitors. Benefiting from the nanostructures, the as-prepared electrode shows a specific capacitance of 413 F g(-1), and no obvious decay even after 3000 charge-discharge cycles. Such a quasi-two-dimensional material is a new kind of supercapacitor electrode material with high performance.

Adult amphibian epidermal proteins: biochemical characterization and developmental appearance.

PubMed

Reeves, O R

1975-08-01

The keratin-like proteins (KLPs) from the epidermis of adult frogs of the species Xenopus laevis have been isolated and biochemically characterized by means of polyacrylamide gel electrophoresis, amino acid analysis, tryptic peptide mapping, amino-terminal end-group analysis and isoelectric focusing. One particular protein fraction of rather unusual amino acid composition found only in epidermal tissue was isolated in quantity by preparative gel electrophoresis and monospecific antibodies prepared against it. Using this anti-KLP antibody preparation it was possible to show that at least one kine of keratin-like protein characteristic of the adult epidermis first appears within the larval epidermis during metamorphosis. This is the first reported biochemical characterization of a tissue-specific protien from adult amphibian skin.

Comparison of arbitrarily primed PCR and macrorestriction (pulsed-field gel electrophoresis) typing of Pseudomonas aeruginosa strains from cystic fibrosis patients.

PubMed Central

Kersulyte, D; Struelens, M J; Deplano, A; Berg, D E

1995-01-01

Arbitrarily primed PCR fingerprinting was carried out on 43 Pseudomonas aeruginosa isolates from cystic fibrosis (CF) patients. Seventeen major groups of strains that coincided with groups also distinguished by macrorestriction (pulsed-field gel electrophoresis) typing were identified. Our results illustrated that a CF patient can carry more than one strain and can carry a given strain for long periods of time and that strains can evolve by changes in drug resistance or other phenotypic traits during long-term colonization. The arbitrarily primed PCR method is recommended for first-pass screening of P. aeruginosa isolates from CF patients, especially when many strains are to be typed, because of its sensitivity and efficiency. PMID:7559985

Comparison of Pulsed-Field Gel Electrophoresis, Ribotyping, and Plasmid Profiling for Typing of Vibrio anguillarum Serovar O1

PubMed Central

Skov, M. N.; Pedersen, K.; Larsen, J. L.

1995-01-01

A total of 75 Vibrio anguillarum serogroup O1 strains were studied with respect to their plasmid contents, ribotypes, and pulsed-field gel electrophoresis (PFGE) patterns. Eight plasmid profiles and six ribotypes were demonstrated, and one profile was dominant by both typing methods. In contrast, PFGE had very high discriminatory power, demonstrating 35 profiles. On the basis of PFGE patterns, a similarity matrix and a dendrogram were constructed. The results indicated that Scandinavian strains and southern European isolates (with some exceptions) belong to two different clonal lineages. A few strains from the United States and United Kingdom deviated considerably from each other and from Scandinavian and southern European strains. PMID:16535003

Studies on a lipopolysaccharide-protein complex from Yersinia pseudotuberculosis. 1 isolation and characterization.

PubMed

Solov'eva, T F; Yermak, I M; Bondarenko, O D; Frolova, G M; Ovodov, Y S

1979-01-01

A comparative study of various procedures of a lipopolysaccharide-protein complex (LPPC) from Yersinia pseudotuberculosis was carried out. The materials obtained were fractionated by molecular-sieve chromatography on Sepharose 2B resulting in highly aggregated complexes with antigen activity. LPPC aggregates dissociated in the presence of sodium dodecylsulphate (SDS) and urea. The chemical composition and serologic properties of fractions obtained are under consideration. The protein component of the complex consists of two major polypeptides (molecular weights--45,000 and 20,000) and some minor ones. The LPS component appeared to give 2--3 narrow bands in gel under conditions of SDS-polyacrylamide gel electrophoresis. It is suggested that such fractionation is caused by LPS association-dissociation in the course of electrophoresis.

Molecular approaches to analysing the microbial composition of raw milk and raw milk cheese.

PubMed

Quigley, Lisa; O'Sullivan, Orla; Beresford, Tom P; Ross, R Paul; Fitzgerald, Gerald F; Cotter, Paul D

2011-11-01

The availability and application of culture-independent tools that enable a detailed investigation of the microbiota and microbial biodiversity of food systems has had a major impact on food microbiology. This review focuses on the application of DNA-based technologies, such as denaturing gradient gel electrophoresis (DGGE), temporal temperature gradient gel electrophoresis (TTGE), single stranded conformation polymorphisms (SSCP), the polymerase chain reaction (PCR) and others, to investigate the diversity, dynamics and identity of microbes in dairy products from raw milk. Here, we will highlight the benefits associated with culture-independent methods which include enhanced sensitivity, rapidity and the detection of microorganisms not previously associated with such products. Copyright © 2011 Elsevier B.V. All rights reserved.

Solution-Processed Ultrathin TiO2 Compact Layer Hybridized with Mesoporous TiO2 for High-Performance Perovskite Solar Cells.

PubMed

Jeong, Inyoung; Park, Yun Hee; Bae, Seunghwan; Park, Minwoo; Jeong, Hansol; Lee, Phillip; Ko, Min Jae

2017-10-25

The electron transport layer (ETL) is a key component of perovskite solar cells (PSCs) and must provide efficient electron extraction and collection while minimizing the charge recombination at interfaces in order to ensure high performance. Conventional bilayered TiO 2 ETLs fabricated by depositing compact TiO 2 (c-TiO 2 ) and mesoporous TiO 2 (mp-TiO 2 ) in sequence exhibit resistive losses due to the contact resistance at the c-TiO 2 /mp-TiO 2 interface and the series resistance arising from the intrinsically low conductivity of TiO 2 . Herein, to minimize such resistive losses, we developed a novel ETL consisting of an ultrathin c-TiO 2 layer hybridized with mp-TiO 2 , which is fabricated by performing one-step spin-coating of a mp-TiO 2 solution containing a small amount of titanium diisopropoxide bis(acetylacetonate) (TAA). By using electron microscopies and elemental mapping analysis, we establish that the optimal concentration of TAA produces an ultrathin blocking layer with a thickness of ∼3 nm and ensures that the mp-TiO 2 layer has a suitable porosity for efficient perovskite infiltration. We compare PSCs based on mesoscopic ETLs with and without compact layers to determine the role of the hole-blocking layer in their performances. The hybrid ETLs exhibit enhanced electron extraction and reduced charge recombination, resulting in better photovoltaic performances and reduced hysteresis of PSCs compared to those with conventional bilayered ETLs.

Evidence of a Transition Layer between the Free Surface and the Bulk.

PubMed

Ogieglo, Wojciech; Tempelman, Kristianne; Napolitano, Simone; Benes, Nieck E

2018-03-15

The free surface, a very thin layer at the interface between polymer and air, is considered the main source of the perturbations in the properties of ultrathin polymer films, i.e., nanoconfinement effects. The structural relaxation of such a layer is decoupled from the molecular dynamics of the bulk. The free surface is, in fact, able to stay liquid even below the temperature where the polymer resides in the glassy state. Importantly, this surface layer is expected to have a very sharp interface with the underlying bulk. Here, by analyzing the penetration of n-hexane into polystyrene films, we report on the existence of a transition region, not observed by previous investigations, extending for 12 nm below the free surface. The presence of such a layer permits reconciling the behavior of interfacial layers with current models and has profound implications on the performance of ultrathin membranes. We show that the expected increase in the flux of the permeating species is actually overruled by nanoconfinement.

The influence of nano-oxide layer on magnetostriction of sensing layer in bottom spin valves

NASA Astrophysics Data System (ADS)

Qiu, J. J.; Han, G. C.; Li, K. B.; Liu, Z. Y.; Zong, B. Y.; Wu, Y. H.

2006-05-01

The magnetostriction coefficient (λs) of ultrathin sputtered polycrystalline as-deposited and annealed Ta/Ni81Fe19(t)/Ta films was studied as a function of the thickness. λs and magnetoresistance (MR) of bottom-type spin valves (SVs) with nano-oxide layer (NOL) added in the pinned layer were investigated by using NiFe, Co90Fe10, and CoFe/NiFe/CoFe layers as free layer (FL), respectively. λs of SV with NOL increased slightly except that of CoFe FL. NOLs were added at different positions to study the effects of NOL on λs of CoFe FL. All λs of CoFe FL change from negative to positive and its absolute value also increases significantly with CoFeOx related NOL added below. Our λs and surface roughness results indicated that the structure of the film not the roughness dominates λs of ultrathin FL in SVs.

Clay induced aggregation of a tetra-cationic metalloporphyrin in Layer by Layer self assembled film

NASA Astrophysics Data System (ADS)

Banik, Soma; Bhattacharjee, J.; Hussain, S. A.; Bhattacharjee, D.

2015-12-01

Porphyrins have a general tendency to form aggregates in ultrathin films. Also electrostatic adsorption of cationic porphyrins onto anionic nano clay platelets results in the flattening of porphyrin moieties. The flattening is evidenced by the red-shifting of Soret band with respect to the aqueous solution. In the present communication, we have studied the clay induced aggregation behaviour of a tetra-cationic metalloporphyrin Manganese (III) 5, 10, 15, 20-tetra (4 pyridyl)-21 H, 23 H-porphine chloride tetrakis (methochloride) (MnTMPyP) in Layer-by-Layer (LbL) self assembled film. The adsorption of dye molecules onto nano clay platelets resulted in the flattening of the meso substituent groups of the dye chromophore. In Layer-by-Layer ultrathin film, the flattened porphyrin molecules tagged nano clay platelets were further associated to form porphyrin aggregates. This has been clearly demonstrated from the UV-vis absorption spectroscopic studies. Atomic Force Microscopic (AFM) studies gave visual evidence of the association of organo-clay hybrid molecules in the LbL film.

Comparative two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of human milk to identify dysregulated proteins in breast cancer.

PubMed

Aslebagh, Roshanak; Channaveerappa, Devika; Arcaro, Kathleen F; Darie, Costel C

2018-05-13

Breast cancer (BC) remains a major cause of mortality, and early detection is considered important for reducing BC-associated deaths. Early detection of BC is challenging in young women, due to the limitations of mammography on the dense breast tissue of young women. We recently reported results of a pilot proteomics study, using one-dimensional polyacrylamide gel electrophoresis (1D-PAGE) and mass spectrometry (MS) to investigate differences in milk proteins from women with and without BC. Here, we applied two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MS to compare the protein pattern in milk from the breasts of a single woman who was diagnosed with BC in one breast 24 months after donating her milk. Statistically different gel spots were picked for protein digestion followed by nanoliquid chromatography tandem MS (nanoLC-MS/MS) analysis. The upregulated proteins in BC versus control are alpha-amylase, gelsolin isoform a precursor, alpha-2-glycoprotein 1 zinc isoform CRA_b partial, apoptosis-inducing factor 2 and vitronectin. Several proteins were downregulated in the milk of the breast later diagnosed with cancer as compared to the milk from the healthy breast, including different isoforms of albumin, cholesterol esterase, different isoforms of lactoferrin, different proteins from the casein family and different isoforms of lysozyme. Results warrant further studies to determine the usefulness of these milk proteins for assessing risk and detecting occult disease. MS data is available via ProteomeXchange with identifier PXD009860. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

High performance p-type organic thin film transistors with an intrinsically photopatternable, ultrathin polymer dielectric layer☆

PubMed Central

Petritz, Andreas; Wolfberger, Archim; Fian, Alexander; Krenn, Joachim R.; Griesser, Thomas; Stadlober, Barbara

2013-01-01

A high-performing bottom-gate top-contact pentacene-based oTFT technology with an ultrathin (25–48 nm) and electrically dense photopatternable polymeric gate dielectric layer is reported. The photosensitive polymer poly((±)endo,exo-bicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid, diphenylester) (PNDPE) is patterned directly by UV-exposure (λ = 254 nm) at a dose typical for conventionally used negative photoresists without the need for any additional photoinitiator. The polymer itself undergoes a photo-Fries rearrangement reaction under UV illumination, which is accompanied by a selective cross-linking of the macromolecules, leading to a change in solubility in organic solvents. This crosslinking reaction and the negative photoresist behavior are investigated by means of sol–gel analysis. The resulting transistors show a field-effect mobility up to 0.8 cm2 V−1 s−1 at an operation voltage as low as −4.5 V. The ultra-low subthreshold swing in the order of 0.1 V dec−1 as well as the completely hysteresis-free transistor characteristics are indicating a very low interface trap density. It can be shown that the device performance is completely stable upon UV-irradiation and development according to a very robust chemical rearrangement. The excellent interface properties, the high stability and the small thickness make the PNDPE gate dielectric a promising candidate for fast organic electronic circuits. PMID:24748853

Rapid Detection of Glycogen Synthase Kinase-3 Activity in Mouse Sperm Using Fluorescent Gel Shift Electrophoresis

PubMed Central

Choi, Hoseok; Choi, Bomi; Seo, Ju Tae; Lee, Kyung Jin; Gye, Myung Chan; Kim, Young-Pil

2016-01-01

Assaying the glycogen synthase kinase-3 (GSK3) activity in sperm is of great importance because it is closely implicated in sperm motility and male infertility. While a number of studies on GSK3 activity have relied on labor-intensive immunoblotting to identify phosphorylated GSK3, here we report the simple and rapid detection of GSK3 activity in mouse sperm using conventional agarose gel electrophoresis and a fluorescent peptide substrate. When a dye-tethered and prephosphorylated (primed) peptide substrate for GSK3 was employed, a distinct mobility shift in the fluorescent bands on the agarose was observed by GSK3-induced phosphorylation of the primed peptides. The GSK3 activity in mouse testes and sperm were quantifiable by gel shift assay with low sample consumption and were significantly correlated with the expression levels of GSK3 and p-GSK3. We suggest that our assay can be used for reliable and rapid detection of GSK3 activity in cells and tissue extracts. PMID:27092510

Theory of DNA electrophoresis in physical gels and entangled polymer solutions

NASA Astrophysics Data System (ADS)

Duke, Thomas; Viovy, Jean Louis

1994-03-01

A scaling theory is presented for the electrophoretic mobility of DNA in sieving media that form dynamically evolving meshworks, such as physical gels and solutions of entangled polymers. In such media, the topological constraints on the DNA's motion are perpetually changing as cross links break and rejoin or as the polymers diffuse. It is shown that if the rate of constraint release falls within a certain range (which depends on the field strength), fractionation can be extended to higher molecular weights than would be feasible using a permanent gel of equivalent pore size. This improvement is a consequence of the disruptive effect that constraint release has on the mechanism of molecular orientation. Numerical simulations support the predictions of the theory. The possibility of realizing such a system in practice, with the aim of improving on current electrophoresis methods, is commented upon. It is suggested that semidilute polymer solutions may be a versatile medium for the rapid separation of long single-stranded DNA molecules, and the particular quality of solution required is identified.

Non-Gradient Blue Native Polyacrylamide Gel Electrophoresis.

PubMed

Luo, Xiaoting; Wu, Jinzi; Jin, Zhen; Yan, Liang-Jun

2017-02-02

Gradient blue native polyacrylamide gel electrophoresis (BN-PAGE) is a well established and widely used technique for activity analysis of high-molecular-weight proteins, protein complexes, and protein-protein interactions. Since its inception in the early 1990s, a variety of minor modifications have been made to this gradient gel analytical method. Here we provide a major modification of the method, which we call non-gradient BN-PAGE. The procedure, similar to that of non-gradient SDS-PAGE, is simple because there is no expensive gradient maker involved. The non-gradient BN-PAGE protocols presented herein provide guidelines on the analysis of mitochondrial protein complexes, in particular, dihydrolipoamide dehydrogenase (DLDH) and those in the electron transport chain. Protocols for the analysis of blood esterases or mitochondrial esterases are also presented. The non-gradient BN-PAGE method may be tailored for analysis of specific proteins according to their molecular weight regardless of whether the target proteins are hydrophobic or hydrophilic. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Investigations on gel forming media use in low gravity bioseparations research

NASA Technical Reports Server (NTRS)

Todd, Paul; Szlag, David C.; Plank, Lindsay D.; Delcourt, Scott G.; Kunze, M. Elaine

1989-01-01

Research on gelling media and conditions suitable for the preservation of the spatial configuration of cell suspensions and macromolecular solutions after separation in free fluid during low gravity experiments is presented. The examples studied included free electrophoresis of cells in a cylindrical column and two-phase aqueous polymer separation. Microgravity electrophoresis experiments were simulated by separating model cell types (animal or human) in a vertical density gradient containing low-conductivity buffer, 1.7-6.5 percent Ficoll, 6.8-5.0 percent sucrose, and 1 percent SeaPrep low-melting temperature agarose. Upon cooling, a gel formed in the column and cells could be captured at the forming locations. Two-phase extraction experiments were simulated using two-polymer solutions in which phase separation occurs in normal saline at temperatures compatible with cell viability and in which one or both phases form a gel upon cooling. Suitable polymers included commercial agaroses (1-2 percent), maltodextrin (5-7 percent), and gelatin (5-20 percent).

Quantitative gel electrophoresis: new records in precision by elaborated staining and detection protocols.

PubMed

Deng, Xi; Schröder, Simone; Redweik, Sabine; Wätzig, Hermann

2011-06-01

Gel electrophoresis (GE) is a very common analytical technique for proteome research and protein analysis. Despite being developed decades ago, there is still a considerable need to improve its precision. Using the fluorescence of Colloidal Coomassie Blue -stained proteins in near-infrared (NIR), the major error source caused by the unpredictable background staining is strongly reduced. This result was generalized for various types of detectors. Since GE is a multi-step procedure, standardization of every single step is required. After detailed analysis of all steps, the staining and destaining were identified as the major source of the remaining variation. By employing standardized protocols, pooled percent relative standard deviations of 1.2-3.1% for band intensities were achieved for one-dimensional separations in repetitive experiments. The analysis of variance suggests that the same batch of staining solution should be used for gels of one experimental series to minimize day-to-day variation and to obtain high precision. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Isoforms of a cuticular protein from larvae of the meal beetle, Tenebrio molitor, studied by mass spectrometry in combination with Edman degradation and two-dimensional polyacrylamide gel electrophoresis.

PubMed Central

Haebel, S.; Jensen, C.; Andersen, S. O.; Roepstorff, P.

1995-01-01

Simultaneous sequencing, using a combination of mass spectrometry and Edman degradation, of three approximately 15-kDa variants of a cuticular protein extracted from the meal beetle Tenebrio molitor larva is demonstrated. The information obtained by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) time-course monitoring of enzymatic digests was found essential to identify the differences among the three variants and for alignment of the peptides in the sequence. To determine whether each individual insect larva contains all three protein variants, proteins extracted from single animals were separated by two-dimensional gel electrophoresis, electroeluted from the gel spots, and analyzed by MALDI MS. Molecular weights of the proteins present in each sample could be obtained, and mass spectrometric mapping of the peptides after digestion with trypsin gave additional information. The protein isoforms were found to be allelic variants. PMID:7795523

Isoforms of a cuticular protein from larvae of the meal beetle, Tenebrio molitor, studied by mass spectrometry in combination with Edman degradation and two-dimensional polyacrylamide gel electrophoresis.

PubMed

Haebel, S; Jensen, C; Andersen, S O; Roepstorff, P

1995-03-01

Simultaneous sequencing, using a combination of mass spectrometry and Edman degradation, of three approximately 15-kDa variants of a cuticular protein extracted from the meal beetle Tenebrio molitor larva is demonstrated. The information obtained by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) time-course monitoring of enzymatic digests was found essential to identify the differences among the three variants and for alignment of the peptides in the sequence. To determine whether each individual insect larva contains all three protein variants, proteins extracted from single animals were separated by two-dimensional gel electrophoresis, electroeluted from the gel spots, and analyzed by MALDI MS. Molecular weights of the proteins present in each sample could be obtained, and mass spectrometric mapping of the peptides after digestion with trypsin gave additional information. The protein isoforms were found to be allelic variants.

2-DE combined with two-layer feature selection accurately establishes the origin of oolong tea.

PubMed

Chien, Han-Ju; Chu, Yen-Wei; Chen, Chi-Wei; Juang, Yu-Min; Chien, Min-Wei; Liu, Chih-Wei; Wu, Chia-Chang; Tzen, Jason T C; Lai, Chien-Chen

2016-11-15

Taiwan is known for its high quality oolong tea. Because of high consumer demand, some tea manufactures mix lower quality leaves with genuine Taiwan oolong tea in order to increase profits. Robust scientific methods are, therefore, needed to verify the origin and quality of tea leaves. In this study, we investigated whether two-dimensional gel electrophoresis (2-DE) and nanoscale liquid chromatography/tandem mass spectroscopy (nano-LC/MS/MS) coupled with a two-layer feature selection mechanism comprising information gain attribute evaluation (IGAE) and support vector machine feature selection (SVM-FS) are useful in identifying characteristic proteins that can be used as markers of the original source of oolong tea. Samples in this study included oolong tea leaves from 23 different sources. We found that our method had an accuracy of 95.5% in correctly identifying the origin of the leaves. Overall, our method is a novel approach for determining the origin of oolong tea leaves. Copyright © 2016 Elsevier Ltd. All rights reserved.

Zinc-decorated silica-coated magnetic nanoparticles for protein binding and controlled release.

PubMed

Bele, Marjan; Hribar, Gorazd; Campelj, Stanislav; Makovec, Darko; Gaberc-Porekar, Vladka; Zorko, Milena; Gaberscek, Miran; Jamnik, Janko; Venturini, Peter

2008-05-01

The aim of this study was to be able to reversibly bind histidine-rich proteins to the surface of maghemite magnetic nanoparticles via coordinative bonding using Zn ions as the anchoring points. We showed that in order to adsorb Zn ions on the maghemite, the surface of the latter needs to be modified. As silica is known to strongly adsorb zinc ions, we chose to modify the maghemite nanoparticles with a nanometre-thick silica layer. This layer appeared to be thin enough for the maghemite nanoparticles to preserve their superparamagnetic nature. As a model the histidine-rich protein bovine serum albumin (BSA) was used. The release of the BSA bound to Zn-decorated silica-coated maghemite nanoparticles was analysed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We demonstrated that the bonding of the BSA to such modified magnetic nanoparticles is highly reversible and can be controlled by an appropriate change of the external conditions, such as a pH decrease or the presence/supply of other chelating compounds.

High Throughput Screen to Identify Novel Drugs that Inhibit Prostate Cancer Metastasis

DTIC Science & Technology

2007-10-01

determined by electrophoresis on 8% denaturing polyacryl - amide gel containing 7 M urea. A 10-bp 32P-labeled ladder and sequencing reaction performed with...isolated from nuclear lysates from P69 and C4-2 cells. The bands shown as rectangles were excised after staining of the proteins in the gel and then...the same primer on a genomic clone was used as a reference. The gel was dried, and the radioactive signals were identified by phos- phorimaging (Storm

Ultraflexible organic amplifier with biocompatible gel electrodes

PubMed Central

Sekitani, Tsuyoshi; Yokota, Tomoyuki; Kuribara, Kazunori; Kaltenbrunner, Martin; Fukushima, Takanori; Inoue, Yusuke; Sekino, Masaki; Isoyama, Takashi; Abe, Yusuke; Onodera, Hiroshi; Someya, Takao

2016-01-01

In vivo electronic monitoring systems are promising technology to obtain biosignals with high spatiotemporal resolution and sensitivity. Here we demonstrate the fabrication of a biocompatible highly conductive gel composite comprising multi-walled carbon nanotube-dispersed sheet with an aqueous hydrogel. This gel composite exhibits admittance of 100 mS cm−2 and maintains high admittance even in a low-frequency range. On implantation into a living hypodermal tissue for 4 weeks, it showed a small foreign-body reaction compared with widely used metal electrodes. Capitalizing on the multi-functional gel composite, we fabricated an ultrathin and mechanically flexible organic active matrix amplifier on a 1.2-μm-thick polyethylene-naphthalate film to amplify (amplification factor: ∼200) weak biosignals. The composite was integrated to the amplifier to realize a direct lead epicardial electrocardiography that is easily spread over an uneven heart tissue. PMID:27125910

Structure and morphology evolution of silica-modified pseudoboehmite aerogels during heat treatment

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

Pakharukova, V.P., E-mail: verapakh@catalysis.ru; Novosibirsk State University, Pirogova Street 2, 630090 Novosibirsk; Research and Educational Center for Energy Efficient Catalysis, Novosibirsk State University, Novosibirsk 630090

Silica-modified pseudoboehmite aerogels (0, 10, 20 at% of Si) were prepared by sol–gel method followed by supercritical drying. The phase transformations, changes in structure and morphology upon calcination were thoroughly investigated by advanced X-Ray diffraction (XRD) techniques and high-resolution transmission electron microscopy (HRTEM). Obtained pseudoboehmite samples had specific nanostructure: ultrathin two-dimensional (2D) crystallites were loosely packed. The silica dopant drastically enhanced the crystallite anisotropy. Thus, the aerogel with Al:Si atomic ratio of 9:1 consisted of the pseudoboehmite nanosheets with thickness of one unit cell (average dimensions of 14.0×1.2×14.5 nm). The specific nanostructure caused remarkable features of experimental XRD patterns, includingmore » anisotropic peak broadening and appearance of forbidden reflection. Direct simulation of XRD patterns with using the Debye Scattering Equation allowed the size and morphology of pseudoboehmite crystallites to be determined. The silica addition strongly delayed formation of γ-alumina and further phase transformations upon calcinaton. Thermal stability of alumina was suggested to be affected by the particle morphology inherited from the pseudoboehmite precursor. - Graphical abstract: Pseudoboehmite samples had specific nanostructure: ultrathin two-dimensional (2D) crystallites were loosely packed. - Highlights: • Silica-doped boehmites were prepared by sol–gel method with supercritical drying. • Ultrathin two-dimensional crystallites of pseudoboehmite were obtained. • Changes in structure and morphology upon calcination were studied. • Simulation of XRD patterns was performed with use of the Debye Scattering Equation. • Thermal stability of alumina depended on morphology inherited from pseudoboehmite.« less

Introducing Students to Protein Analysis Techniques: Separation and Comparative Analysis of Gluten Proteins in Various Wheat Strains

ERIC Educational Resources Information Center

Pirinelli, Alyssa L.; Trinidad, Jonathan C.; Pohl, Nicola L. B.

2016-01-01

Polyacrylamide gel electrophoresis (PAGE) is commonly taught in undergraduate laboratory classes as a traditional method to analyze proteins. An experiment has been developed to teach these basic protein gel skills in the context of gluten protein isolation from various types of wheat flour. A further goal is to relate this technique to current…

Production and Characterization of an Avian Ricin Antitoxin

DTIC Science & Technology

1992-01-15

naturally -occurring plant and/or bacterial toxins as biological threat agents, effective antitoxins are needed for either piophylactic or causal...system, an avian antitoxin against the potent phytotoxin , ricin. will be developed and evaluated. The production of therapeutic antibodies in avian...Dynatech). PolyacrylmIde gel electrophoresis (PAGE): Acrylamide gels were prepared according to methods described by Laemmli ( Nature . 227. 1970) and

Tunable separations based on a molecular size effect for biomolecules by poly(ethylene glycol) gel-based capillary electrophoresis.

PubMed

Kubo, Takuya; Nishimura, Naoki; Furuta, Hayato; Kubota, Kei; Naito, Toyohiro; Otsuka, Koji

2017-11-10

We report novel capillary gel electrophoresis (CGE) with poly(ethylene glycol) (PEG)-based hydrogels for the effective separations of biomolecules containing sugars and DNAs based on a molecular size effect. The gel capillaries were prepared in a fused silica capillary modified with 3-(trimethoxysilyl)propylmethacrylate using a variety of the PEG-based hydrogels. After the fundamental evaluations in CGE regarding the separation based on the molecular size effect depending on the crosslinking density, the optimized capillary provided the efficient separation of glucose ladder (G1 to G20). In addition, another capillary showed the successful separation of DNA ladder in the range of 10-1100 base pair, which is superior to an authentic acrylamide-based gel capillary. For both glucose and DNA ladders, the separation ranges against the molecular size were simply controllable by alteration of the concentration and/or units of ethylene oxide in the PEG-based crosslinker. Finally, we demonstrated the separations of real samples, which included sugars carved out from monoclonal antibodies, mAbs, and then the efficient separations based on the molecular size effect were achieved. Copyright © 2017 Elsevier B.V. All rights reserved.

Evaluation of the electroosmotic medium pump system for preparative disk gel electrophoresis.

PubMed

Hayakawa, M; Hosogi, Y; Takiguchi, H; Saito, S; Shiroza, T; Shibata, Y; Hiratsuka, K; Kiyama-Kishikawa, M; Abiko, Y

2001-01-15

This paper describes an improved electroosmotic elution system for preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) based on the epochal idea of H. V. Tan et al. (Nucleic Acids Res. 1988, 16, 1921-1930). In this elution system, a semipermeable membrane, mounted under the gel terminal end, works as the elution pump as well as the partition of the elution chamber. We refer to this system as the "electroosmotic medium pump system." Operation of the constructed apparatus (3.6 cm i.d. disk gel column) and resolution of the protein bands were examined by separation of the model protein mixture (bovine serum albumin (BSA), ovalbumin, bovine carbonic anhydrase, soybean trypsin inhibitor) and purification of the membrane protein, dipeptidyl peptidase IV (DPP IV). The Spectra/Por 7 dialysis membrane provided a better flow profile for the elution buffer. The four model proteins of the protein mixture were able to be completely separated from each other and recovered without dilution. The maximum protein concentration of eluate achieved was 93 mg/ml, when applying a single component, BSA fraction V, as a sample. Furthermore, the multifunctional ectoenzyme, DPP IV, was purified in a single step. Copyright 2001 Academic Press.

Analysis of metal-binding proteins separated by non-denaturating gel electrophoresis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

PubMed

Becker, J Susanne; Mounicou, Sandra; Zoriy, Miroslav V; Becker, J Sabine; Lobinski, Ryszard

2008-09-15

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) have become established as very efficient and sensitive biopolymer and elemental mass spectrometric techniques for studying metal-binding proteins (metalloproteins) in life sciences. Protein complexes present in rat tissues (liver and kidney) were separated in their native state in the first dimension by blue native gel electrophoresis (BN-PAGE). Essential and toxic metals, such as zinc, copper, iron, nickel, chromium, cadmium and lead, were detected by scanning the gel bands using quadrupole LA-ICP-MS with and without collision cell as a microanalytical technique. Several proteins were identified by using MALDI-TOF-MS together with a database search. For example, on one protein band cut from the BN-PAGE gel and digested with the enzyme trypsin, two different proteins - protein FAM44B and cathepsin B precursor - were identified. By combining biomolecular and elemental mass spectrometry, it was possible to characterize and identify selected metal-binding rat liver and kidney tissue proteins.

White organic light-emitting diodes based on doped and ultrathin Rubrene layer

NASA Astrophysics Data System (ADS)

Li, Yi; Jiang, Yadong; Wen, Wen; Yu, Junsheng

2010-10-01

Based on a yellow fluorescent dye of 5, 6, 11, 12-tetraphenylnaphthacene (Rubrene), WOLEDs were fabricated, with doping structure and ultrathin layer structure utilized in the devices. By doping Rubrene into blue-emitting N,N'-bis-(1- naphthyl)-N,N'-biphenyl-1,1'-biphenyl-4,4'-diamine (NPB), the device with a structure of indium-tin-oxide (ITO)/NPB (40 nm)/NPB:Rubrene (0.25 wt%, 7 nm)/2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline (BCP) (30 nm)/Mg:Ag exhibited a warm white light with Commissions Internationale De L'Eclairage (CIE) coordinates of (0.38, 0.41) at 12 V. The electroluminescent spectrum of the OLED consisted of blue and yellow fluorescent emissions, the intensity of blue emission increased gradually relative to the orange emission with increasing voltage. This is mainly due to the recombination zone shifted towards the anode side as the transmission rate of electrons grows faster than that of holes under higher bias voltage. A maximum luminance of 7300 cd/m2 and a maximum power efficiency of 0.57 lm/W were achieved. Comparatively, by utilizing ultrathin dopant layer, the device with a structure of ITO/NPB (40 nm)/Rubrene (0.3 nm)/NPB (7 nm)/BCP (30 nm)/Mg:Ag achieved a low turn-on voltage of 3 V and a more stable white light. The peaks of EL spectra located at 430 and 560 nm corresponding to the CIE coordinates of (0.32, 0.32) under bias voltage ranging from 5 to 15 V. A maximum luminance of 5630 cd/m2 and a maximum power efficiency of 0.6 lm/W were achieved. The balanced spectra were attributed to the stable confining of charge carriers and exciton by the thin emitting layers. Hence, with simple device structure and fabricating process, the device with ultrathin layer achieved low turn-on voltage, stable white light emitting and higher power efficiency.

Detection of Macromolecular Fractions in HCN Polymers Using Electrophoretic and Ultrafiltration Techniques.

PubMed

Marín-Yaseli, Margarita R; Cid, Cristina; Yagüe, Ana I; Ruiz-Bermejo, Marta

2017-02-01

Elucidating the origin of life involves synthetic as well as analytical challenges. Herein, for the first time, we describe the use of gel electrophoresis and ultrafiltration to fractionate HCN polymers. Since the first prebiotic synthesis of adenine by Oró, HCN polymers have gained much interest in studies on the origins of life due to the identification of biomonomers and related compounds within them. Here, we demonstrate that macromolecular fractions with electrophoretic mobility can also be detected within HCN polymers. The migration of polymers under the influence of an electric field depends not only on their sizes (one-dimensional electrophoresis) but also their different isoelectric points (two-dimensional electrophoresis, 2-DE). The same behaviour was observed for several macromolecular fractions detected in HCN polymers. Macromolecular fractions with apparent molecular weights as high as 250 kDa were detected by tricine-SDS gel electrophoresis. Cationic macromolecular fractions with apparent molecular weights as high as 140 kDa were also detected by 2-DE. The HCN polymers synthesized were fractionated by ultrafiltration. As a result, the molecular weight distributions of the macromolecular fractions detected in the HCN polymers directly depended on the synthetic conditions used to produce these polymers. The implications of these results for prebiotic chemistry will be discussed. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

2D-DIGE in Proteomics.

PubMed

Pasquali, Matias; Serchi, Tommaso; Planchon, Sebastien; Renaut, Jenny

2017-01-01

The two-dimensional difference gel electrophoresis method is a valuable approach for proteomics. The method, using cyanine fluorescent dyes, allows the co-migration of multiple protein samples in the same gel and their simultaneous detection, thus reducing experimental and analytical time. 2D-DIGE, compared to traditional post-staining 2D-PAGE protocols (e.g., colloidal Coomassie or silver nitrate), provides faster and more reliable gel matching, limiting the impact of gel to gel variation, and allows also a good dynamic range for quantitative comparisons. By the use of internal standards, it is possible to normalize for experimental variations in spot intensities and gel patterns. Here we describe the experimental steps we follow in our routine 2D-DIGE procedure that we then apply to multiple biological questions.

Monolayer-Mediated Growth of Organic Semiconductor Films with Improved Device Performance.

PubMed

Huang, Lizhen; Hu, Xiaorong; Chi, Lifeng

2015-09-15

Increased interest in wearable and smart electronics is driving numerous research works on organic electronics. The control of film growth and patterning is of great importance when targeting high-performance organic semiconductor devices. In this Feature Article, we summarize our recent work focusing on the growth, crystallization, and device operation of organic semiconductors intermediated by ultrathin organic films (in most cases, only a monolayer). The site-selective growth, modified crystallization and morphology, and improved device performance of organic semiconductor films are demonstrated with the help of the inducing layers, including patterned and uniform Langmuir-Blodgett monolayers, crystalline ultrathin organic films, and self-assembled polymer brush films. The introduction of the inducing layers could dramatically change the diffusion of the organic semiconductors on the surface and the interactions between the active layer with the inducing layer, leading to improved aggregation/crystallization behavior and device performance.

Ultrathin layered double hydroxide nanosheets with Ni(III) active species obtained by exfoliation for highly efficient ethanol electrooxidation

PubMed Central

Xu, Liang; Wang, Zhe; Chen, Xu; Qu, Zongkai; Li, Feng; Yang, Wensheng

2018-01-01

The development of non-precious metal electrocatalysts for renewable energy conversion and storage is compelling but greatly challenging due to low activity of the existing catalysts. Herein, the ultrathin NiAl-layered double hydroxide nanosheets (NiAl-LDH-NSs) are prepared by simple liquid-exfoliation of bulk NiAl-LDHs and first used as ethanol electrooxidation catalysts. The ultrathin two-dimensional (2D) structure ensures that the LDH nanosheets expose a greater number of active sites. More importantly, much Ni(III) active species (NiOOH) in the ultrathin nanosheets are formed by the exfoliation process, which play an authentic catalytic role in the ethanol oxidation reaction (EOR). The presence of NiOOH remarkably improves the reactivity and electrical conductivity of LDH nanosheets. These synergistic effects lead to strikingly more than 30 times enhanced EOR activity of NiAl-LDH-NSs compared to bulk NiAl-LDHs. The obtained electrocatalytic activity is also much better than those of most Ni- and LDH-based EOR catalysts reported to date. In addition, the ultrathin NiAl-LDH-NS electrocatalyst also exhibits good long-term stability (maintain 81.8% of the original value after 10000 s). This study not only provides a highly competitive EOR catalyst, but also opens new avenues toward the design of highly efficient electrode materials that have various potential applications in supercapacitor, Ni-MH battery and other electrocatalytic systems. PMID:29622818

Differential Single Nucleotide Polymorphism-Based Analysis of an Outbreak Caused by Salmonella enterica Serovar Manhattan Reveals Epidemiological Details Missed by Standard Pulsed-Field Gel Electrophoresis

PubMed Central

Scaltriti, Erika; Sassera, Davide; Comandatore, Francesco; Morganti, Marina; Mandalari, Carmen; Gaiarsa, Stefano; Bandi, Claudio; Zehender, Gianguglielmo; Bolzoni, Luca; Casadei, Gabriele

2015-01-01

We retrospectively analyzed a rare Salmonella enterica serovar Manhattan outbreak that occurred in Italy in 2009 to evaluate the potential of new genomic tools based on differential single nucleotide polymorphism (SNP) analysis in comparison with the gold standard genotyping method, pulsed-field gel electrophoresis. A total of 39 isolates were analyzed from patients (n = 15) and food, feed, animal, and environmental sources (n = 24), resulting in five different pulsed-field gel electrophoresis (PFGE) profiles. Isolates epidemiologically related to the outbreak clustered within the same pulsotype, SXB_BS.0003, without any further differentiation. Thirty-three isolates were considered for genomic analysis based on different sets of SNPs, core, synonymous, nonsynonymous, as well as SNPs in different codon positions, by Bayesian and maximum likelihood algorithms. Trees generated from core and nonsynonymous SNPs, as well as SNPs at the second and first plus second codon positions detailed four distinct groups of isolates within the outbreak pulsotype, discriminating outbreak-related isolates of human and food origins. Conversely, the trees derived from synonymous and third-codon-position SNPs clustered food and human isolates together, indicating that all outbreak-related isolates constituted a single clone, which was in line with the epidemiological evidence. Further experiments are in place to extend this approach within our regional enteropathogen surveillance system. PMID:25653407

Differential single nucleotide polymorphism-based analysis of an outbreak caused by Salmonella enterica serovar Manhattan reveals epidemiological details missed by standard pulsed-field gel electrophoresis.

PubMed

Scaltriti, Erika; Sassera, Davide; Comandatore, Francesco; Morganti, Marina; Mandalari, Carmen; Gaiarsa, Stefano; Bandi, Claudio; Zehender, Gianguglielmo; Bolzoni, Luca; Casadei, Gabriele; Pongolini, Stefano

2015-04-01

We retrospectively analyzed a rare Salmonella enterica serovar Manhattan outbreak that occurred in Italy in 2009 to evaluate the potential of new genomic tools based on differential single nucleotide polymorphism (SNP) analysis in comparison with the gold standard genotyping method, pulsed-field gel electrophoresis. A total of 39 isolates were analyzed from patients (n=15) and food, feed, animal, and environmental sources (n=24), resulting in five different pulsed-field gel electrophoresis (PFGE) profiles. Isolates epidemiologically related to the outbreak clustered within the same pulsotype, SXB_BS.0003, without any further differentiation. Thirty-three isolates were considered for genomic analysis based on different sets of SNPs, core, synonymous, nonsynonymous, as well as SNPs in different codon positions, by Bayesian and maximum likelihood algorithms. Trees generated from core and nonsynonymous SNPs, as well as SNPs at the second and first plus second codon positions detailed four distinct groups of isolates within the outbreak pulsotype, discriminating outbreak-related isolates of human and food origins. Conversely, the trees derived from synonymous and third-codon-position SNPs clustered food and human isolates together, indicating that all outbreak-related isolates constituted a single clone, which was in line with the epidemiological evidence. Further experiments are in place to extend this approach within our regional enteropathogen surveillance system. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

2-D Difference in gel electrophoresis combined with Pro-Q Diamond staining: a successful approach for the identification of kinase/phosphatase targets.

PubMed

Orsatti, Laura; Forte, Eleonora; Tomei, Licia; Caterino, Marianna; Pessi, Antonello; Talamo, Fabio

2009-07-01

The protein tyrosine phosphatase PRL-3 is an appealing therapeutic cancer target for its well described involvement in the metastasis progression. Nevertheless, very little is known about PRL-3 role in tumorigenesis. In the attempt to identify the protein target of this phosphatase we have devised a model system based on the use of highly invasive HCT116 colon cancer cells over-expressing PRL-3. We used 2-D difference gel electrophoresis combined with the fluorescence staining Pro-Q Diamond selective for phosphorylated proteins to monitor changes in the phosphorylation status of possible substrates. Proteins whose phosphorylation level was negatively affected by PRL-3 over-expression were identified by MS. Two proteins were found to be significantly dephosphorylated in this condition, the cytoskeletal protein ezrin and elongation factor 2. Ezrin has already been described as having a proactive role in cancer metastasis through control of its phosphorylation status, and the PRL-3-induced modulation of ezrin phosphorylation in HCT116 and human umblical vascular endothelial cells is the subject of a separate paper by Forte et al. [Biochim. Biophys. Acta 2008, 1783, 334-344]. The combination of 2-D difference in gel electrophoresis and Pro-Q Diamond was hence confirmed successful in analyzing changes of protein phosphorylation which enable the identification of kinase/phosphatase targets.

A Comprehensive Quality Evaluation System for Complex Herbal Medicine Using PacBio Sequencing, PCR-Denaturing Gradient Gel Electrophoresis, and Several Chemical Approaches

PubMed Central

Zheng, Xiasheng; Zhang, Peng; Liao, Baosheng; Li, Jing; Liu, Xingyun; Shi, Yuhua; Cheng, Jinle; Lai, Zhitian; Xu, Jiang; Chen, Shilin

2017-01-01

Herbal medicine is a major component of complementary and alternative medicine, contributing significantly to the health of many people and communities. Quality control of herbal medicine is crucial to ensure that it is safe and sound for use. Here, we investigated a comprehensive quality evaluation system for a classic herbal medicine, Danggui Buxue Formula, by applying genetic-based and analytical chemistry approaches to authenticate and evaluate the quality of its samples. For authenticity, we successfully applied two novel technologies, third-generation sequencing and PCR-DGGE (denaturing gradient gel electrophoresis), to analyze the ingredient composition of the tested samples. For quality evaluation, we used high performance liquid chromatography assays to determine the content of chemical markers to help estimate the dosage relationship between its two raw materials, plant roots of Huangqi and Danggui. A series of surveys were then conducted against several exogenous contaminations, aiming to further access the efficacy and safety of the samples. In conclusion, the quality evaluation system demonstrated here can potentially address the authenticity, quality, and safety of herbal medicines, thus providing novel insight for enhancing their overall quality control. Highlight: We established a comprehensive quality evaluation system for herbal medicine, by combining two genetic-based approaches third-generation sequencing and DGGE (denaturing gradient gel electrophoresis) with analytical chemistry approaches to achieve the authentication and quality connotation of the samples. PMID:28955365

A Comprehensive Quality Evaluation System for Complex Herbal Medicine Using PacBio Sequencing, PCR-Denaturing Gradient Gel Electrophoresis, and Several Chemical Approaches.

PubMed

Zheng, Xiasheng; Zhang, Peng; Liao, Baosheng; Li, Jing; Liu, Xingyun; Shi, Yuhua; Cheng, Jinle; Lai, Zhitian; Xu, Jiang; Chen, Shilin

2017-01-01

Herbal medicine is a major component of complementary and alternative medicine, contributing significantly to the health of many people and communities. Quality control of herbal medicine is crucial to ensure that it is safe and sound for use. Here, we investigated a comprehensive quality evaluation system for a classic herbal medicine, Danggui Buxue Formula, by applying genetic-based and analytical chemistry approaches to authenticate and evaluate the quality of its samples. For authenticity, we successfully applied two novel technologies, third-generation sequencing and PCR-DGGE (denaturing gradient gel electrophoresis), to analyze the ingredient composition of the tested samples. For quality evaluation, we used high performance liquid chromatography assays to determine the content of chemical markers to help estimate the dosage relationship between its two raw materials, plant roots of Huangqi and Danggui. A series of surveys were then conducted against several exogenous contaminations, aiming to further access the efficacy and safety of the samples. In conclusion, the quality evaluation system demonstrated here can potentially address the authenticity, quality, and safety of herbal medicines, thus providing novel insight for enhancing their overall quality control. Highlight : We established a comprehensive quality evaluation system for herbal medicine, by combining two genetic-based approaches third-generation sequencing and DGGE (denaturing gradient gel electrophoresis) with analytical chemistry approaches to achieve the authentication and quality connotation of the samples.

Agarose-gel electrophoresis for the quality assurance and purity of heparin formulations.

PubMed

Volpi, Nicola; Buzzega, Dania

2012-01-01

The adulteration of raw heparin (Hep) with a synthetic oversulfated chondroitin sulfate (OSCS) not found in nature produced in 2007-2008 a global crisis giving rise to the development of additional, new and specific methods for its quality assurance and purity. In this study, a simple and sensitive agarose-gel electrophoresis method has been developed for the visualization of OSCS in Hep samples along with other natural glycosaminoglycans possibly present as "process-related impurities", in particular dermatan sulfate (DS) and chondroitin sulfate (CS). Agarose-gel electrophoresis under non-conventional conditions is able to separate OSCS from Hep with its two components, the slow-moving and fast-moving species, DS and CS by performing separation for 15 h (overnight) and under high voltage (100 mA, ∼200 V). Densitometric scanning enabled us to calculate a limit of detection of ∼0.5 μg OSCS with a linear behaviour from 0.1 to 5 μg, comparable to CS/DS. Contaminated samples from Hep manufacturers were analyzed and quantitative data were found comparable to previous studies. Due to its capacity to process many samples in a single run and to the equipment commonly available in laboratories, this analytical method would be suitable for the identification and quantification of contamination by other polysaccharides, in particular OSCS and DS, within Hep preparations and formulations. Copyright © 2012 Elsevier B.V. All rights reserved.

Analysis of polypeptide composition and antigenic components of Rickettsia tsutsugamushi by polyacrylamide gel electrophoresis and immunoblotting.

PubMed Central

Tamura, A; Ohashi, N; Urakami, H; Takahashi, K; Oyanagi, M

1985-01-01

Polyacrylamide gel electrophoresis of lysates of purified Rickettsia tsutsugamushi revealed as many as 30 polypeptide bands, including major bands corresponding to molecular sizes of 70, 60, 54 to 56, and 46 to 47 kilodaltons. Compared with the polypeptide composition of the rickettsiae of Gilliam, Karp, and Kato strains and a newly isolated Shimokoshi strain, the major polypeptide in the Kato strain (54-56K) and in the Karp strain (46-47K) migrated a little faster and slower, respectively, than the corresponding polypeptides in the other strains. The largest major polypeptide (54-56K) was digestible by the treatment of intact rickettsiae with trypsin and variable in content in separate preparations, suggesting that the polypeptide exists on the rickettsial surface and is easily degraded during the handling of these microorganisms. Several surface polypeptides of rickettsiae, including the 54-56K and 46-47K polypeptides, were detected by radioiodination of intact rickettsiae followed by polyacrylamide gel electrophoresis of the lysate; however, the 70K and 60K polypeptides were not labeled. Immunoblotting experiments with hyperimmune sera prepared in guinea pigs against each strain demonstrated that the 70K, 54-56K, and 46-47K polypeptides showed antigenic activities. The 54-56K polypeptide appeared to be strain specific, whereas the 70K and 46-47K polypeptides cross-reacted with the heterologous antisera. Images PMID:3922893

Detection by denaturing gradient gel electrophoresis of ammonia-oxidizing bacteria in microcosms of crude oil-contaminated mangrove sediments.

PubMed

dos Santos, A C F; Marques, E L S; Gross, E; Souza, S S; Dias, J C T; Brendel, M; Rezende, R P

2012-01-27

Currently, the effect of crude oil on ammonia-oxidizing bacterium communities from mangrove sediments is little understood. We studied the diversity of ammonia-oxidizing bacteria in mangrove microcosm experiments using mangrove sediments contaminated with 0.1, 0.5, 1, 2, and 5% crude oil as well as non-contaminated control and landfarm soil from near an oil refinery in Camamu Bay in Bahia, Brazil. The evolution of CO(2) production in all crude oil-contaminated microcosms showed potential for mineralization. Cluster analysis of denaturing gradient gel electrophoresis-derived samples generated with primers for gene amoA, which encodes the functional enzyme ammonia monooxygenase, showed differences in the sample contaminated with 5% compared to the other samples. Principal component analysis showed divergence of the non-contaminated samples from the 5% crude oil-contaminated sediment. A Venn diagram generated from the banding pattern of PCR-denaturing gradient gel electrophoresis was used to look for operational taxonomic units (OTUs) in common. Eight OTUs were found in non-contaminated sediments and in samples contaminated with 0.5, 1, or 2% crude oil. A Jaccard similarity index of 50% was found for samples contaminated with 0.1, 0.5, 1, and 2% crude oil. This is the first study that focuses on the impact of crude oil on the ammonia-oxidizing bacterium community in mangrove sediments from Camamu Bay.

Ultra-thin enhanced-absorption long-wave infrared detectors

NASA Astrophysics Data System (ADS)

Wang, Shaohua; Yoon, Narae; Kamboj, Abhilasha; Petluru, Priyanka; Zheng, Wanhua; Wasserman, Daniel

2018-02-01

We propose an architecture for enhanced absorption in ultra-thin strained layer superlattice detectors utilizing a hybrid optical cavity design. Our detector architecture utilizes a designer-metal doped semiconductor ground plane beneath the ultra-subwavelength thickness long-wavelength infrared absorber material, upon which we pattern metallic antenna structures. We demonstrate the potential for near 50% detector absorption in absorber layers with thicknesses of approximately λ0/50, using realistic material parameters. We investigate detector absorption as a function of wavelength and incidence angle, as well as detector geometry. The proposed device architecture offers the potential for high efficiency detectors with minimal growth costs and relaxed design parameters.

A FRET system built on quartz plate as a ratiometric fluorescence sensor for mercury ions in water.

PubMed

Liu, Baoyu; Zeng, Fang; Liu, Yan; Wu, Shuizhu

2012-04-07

Due to the hazardous nature of mercury ions, the development of a cost effective, sensitive and field-portable sensor is of high significance for both industry and civilian use. In this work, a FRET-based ratiometric sensor for detecting mercury ions in water was fabricated by depositing a multilayered silica structure on a quartz plate. For the preparation of the film-based sensor, a silica support layer was first deposited on the quartz plate by using the sol-gel spin-coating procedure, and three ultrathin functional layers (donor, spacer and receptor) were then deposited on the support layer by dip-coating in a stepwise manner in toluene solution. As the film-based sensor was placed into an aqueous solution of Hg(2+), the non-fluorescent receptor (a spirolactam rhodamine derivative) on the film surface could form a complex with the mercury ion and act as the acceptor of the energy transfer. Upon excitation, the donor (a nitrobenzoxadiazolyl derivative, NBD) could transfer its excited energy from the donor layer to the acceptor on the film surface via the 'through space' energy transfer process, thus realizing the FRET-based ratiometric sensing for mercury ions. The sensor can selectively detect Hg(2+) in water with the detection limit of 1 μM. This solid film sensor is capable of being easily-portable and visualized detection. This strategy may offer new approaches for constructing other FRET-based solid-state devices.

Tuning the ultrafast photodissociation dynamics of CH3Br on ultrathin MgO films by reducing the layer thickness to the 2D limit

NASA Astrophysics Data System (ADS)

Vaida, Mihai E.; Bernhardt, Thorsten M.

2017-11-01

The femtosecond-laser induced photodissociation of CH3Br adsorbed at sub-monolayer coverage on a solid surface was investigated by time-resolved pump-probe mass spectrometry. To tune the interaction of the CH3Br molecules with the substrate, an Mo(1 0 0) surface was covered with ultrathin insulating MgO layers of variable thickness. By gradually decreasing the magnesia layer thickness to the 2D limit the photodissociation dynamics observed by detection of the methyl fragment indicates an energetic lowering of the relevant methyl bromide excited states due to the increasing spatial proximity of the metallic support. Potential orientational effects of the methyl bromide adsorption geometry are also considered.

Exchange bias and perpendicular anisotropy study of ultrathin Pt-Co-Pt-IrMn multilayers sputtered on float glass

NASA Astrophysics Data System (ADS)

Laval, M.; Lüders, U.; Bobo, J. F.

2007-09-01

We have prepared ultrathin Pt-Co-Pt-IrMn polycrystalline multilayers on float-glass substrates by DC magnetron sputtering. We have determined the optimal set of thickness for both Pt layers, the Co layer and the IrMn biasing layer so that these samples exhibit at the same time out-of-plane magnetic anisotropy and exchange bias. Kerr microscopy domain structure imaging evidences an increase of nucleation rate accompanied with inhomogeneous magnetic behavior in the case of exchange-biased films compared to Pt-Co-Pt trilayers. Polar hysteresis loops are measured in obliquely applied magnetic field conditions, allowing us to determine both perpendicular anisotropy effective constant Keff and exchange-bias coupling JE, which are significantly different from the ones determined by standard switching field measurements.

Anatase TiO2 ultrathin nanobelts derived from room-temperature-synthesized titanates for fast and safe lithium storage

PubMed Central

Wen, Wei; Wu, Jin-ming; Jiang, Yin-zhu; Yu, Sheng-lan; Bai, Jun-qiang; Cao, Min-hua; Cui, Jie

2015-01-01

Lithium-ion batteries (LIBs) are promising energy storage devices for portable electronics, electric vehicles, and power-grid applications. It is highly desirable yet challenging to develop a simple and scalable method for constructions of sustainable materials for fast and safe LIBs. Herein, we exploit a novel and scalable route to synthesize ultrathin nanobelts of anatase TiO2, which is resource abundant and is eligible for safe anodes in LIBs. The achieved ultrathin nanobelts demonstrate outstanding performances for lithium storage because of the unique nanoarchitecture and appropriate composition. Unlike conventional alkali-hydrothermal approaches to hydrogen titanates, the present room temperature alkaline-free wet chemistry strategy guarantees the ultrathin thickness for the resultant titanate nanobelts. The anatase TiO2 ultrathin nanobelts were achieved simply by a subsequent calcination in air. The synthesis route is convenient for metal decoration and also for fabricating thin films of one/three dimensional arrays on various substrates at low temperatures, in absence of any seed layers. PMID:26133276

A versatile electrophoresis system for the analysis of high- and low-molecular-weight proteins

PubMed Central

Tastet, Christophe; Lescuyer, Pierre; Diemer, Hélène; Luche, Sylvie; van Dorsselaer, Alain; Rabilloud, Thierry

2003-01-01

A new, versatile, multiphasic buffer system for high resolution sodium dodecyl sulfatepolyacrylamide gel electrophoresis of proteins in the relative molecular weight Mw range of 300,000-3000 Da is described. The system, based on the theory of multiphasic zone electrophoresis, allows complete stacking and destacking of proteins in the above Mw range. The buffer system uses taurine and chloride as trailing and leading ion, respectively, and Tris, at a pH close to its pKa, as the buffering counter ion. Coupled with limited variation in the acrylamide concentration, this electrophoresis system allows to tailor the resolution in the 6–200 kDa Mw range, with minimal difficulties in the post electrophoretic identification processes. PMID:12783456

Effects of near-UV radiation on the protein of the grey squirrel lens.

PubMed

Zigman, S; Paxhia, T; Waldron, W

1988-06-01

In vivo exposure of grey squirrels to 40W BLB illumination resulted in alterations in the state of the lens crystallins, mainly in the outer layer of the lens. HPLC revealed an increase of the void volume or crosslinked crystallins and an increase in peptides with molecular weights lower than 20,000 d. In vitro exposure of squirrel lens aqueous extracts to Woods lamp radiation (predominantly 365 nm) led to similar but more exaggerated changes as viewed by high performance liquid chromatography. When viewed by polyacrylamide gel electrophoresis (PAGE), soluble protein crosslinking was also observed. The near-UV absorbing chromophores of low molecular weight present in the lens served as photosensitizers that enhanced the protein changes. Sodium azide inhibited the changes, indicating a role for singlet oxygen in the crosslinking.

Facile fabrication of ultrathin Pt overlayers onto nanoporous metal membranes via repeated Cu UPD and in situ redox replacement reaction.

PubMed

Liu, Pengpeng; Ge, Xingbo; Wang, Rongyue; Ma, Houyi; Ding, Yi

2009-01-06

Ultrathin Pt films from one to several atomic layers are successfully decorated onto nanoporous gold (NPG) membranes by utilizing under potential deposition (UPD) of Cu onto Au or Pt surfaces, followed by in situ redox replacement reaction (RRR) of UPD Cu by Pt. The thickness of Pt layers can be controlled precisely by repeating the Cu-UPD-RRR cycles. TEM observations coupled with electrochemical testing suggest that the morphology of Pt overlayers changes from an ultrathin epitaxial film in the case of one or two atomic layers to well-dispersed nanoislands in the case of four and more atomic layers. Electron diffraction (ED) patterns confirm that the as-prepared NPG-Pt membranes maintain a single-crystalline structure, even though the thickness of Pt films reaches six atomic layers, indicating the decorated Pt films hold the same crystallographic relationship to the NPG substrate during the entire fabrication process. Due to the regular modulation of Pt utilization, the electrocatalytic activity of NPG-Pt exhibits interesting surface structure dependence in methanol, ethanol, and CO electrooxidation reactions. These novel bimetallic nanocatalysts show excellent electrocatalytic activity and much enhanced poison tolerance as compared to the commercial Pt/C catalysts. The success in the fabrication of NPG-Pt-type materials provides a new path to prepare electrocatalysts with ultralow Pt loading and high Pt utilization, which is of great significance in energy-related applications, such as direct alcohol fuel cells (DAFCs).

Electric field effect on exchange interaction in ultrathin Co films with ionic liquids

NASA Astrophysics Data System (ADS)

Ishibashi, Mio; Yamada, Kihiro T.; Shiota, Yoichi; Ando, Fuyuki; Koyama, Tomohiro; Kakizakai, Haruka; Mizuno, Hayato; Miwa, Kazumoto; Ono, Shimpei; Moriyama, Takahiro; Chiba, Daichi; Ono, Teruo

2018-06-01

Electric-field modulations of magnetic properties have been extensively studied not only for practical applications but also for fundamental interest. In this study, we investigated the electric field effect on the exchange interaction in ultrathin Co films with ionic liquids. The exchange coupling J was characterized from the direct magnetization measurement as a function of temperature using Pt/ultrathin Co/MgO structures. The trend of the electric field effect on J is in good agreement with that of the theoretical prediction, and a large change in J by applying a gate voltage was observed by forming an electric double layer using ionic liquids.

Ultrathin ZnS and ZnO Interfacial Passivation Layers for Atomic-Layer-Deposited HfO2 Films on InP Substrates.

PubMed

Kim, Seung Hyun; Joo, So Yeong; Jin, Hyun Soo; Kim, Woo-Byoung; Park, Tae Joo

2016-08-17

Ultrathin ZnS and ZnO films grown by atomic layer deposition (ALD) were employed as interfacial passivation layers (IPLs) for HfO2 films on InP substrates. The interfacial layer growth during the ALD of the HfO2 film was effectively suppressed by the IPLs, resulting in the decrease of electrical thickness, hysteresis, and interface state density. Compared with the ZnO IPL, the ZnS IPL was more effective in reducing the interface state density near the valence band edge. The leakage current density through the film was considerably lowered by the IPLs because the film crystallization was suppressed. Especially for the film with the ZnS IPL, the leakage current density in the low-voltage region was significantly lower than that observed for the film with the ZnO IPL, because the direct tunneling current was suppressed by the higher conduction band offset of ZnS with the InP substrate.

Ultrathin nitrogen-doped graphitized carbon shell encapsulating CoRu bimetallic nanoparticles for enhanced electrocatalytic hydrogen evolution.

PubMed

Xu, You; Li, Yinghao; Yin, Shuli; Yu, Hongjie; Xue, Hairong; Li, Xiaonian; Wang, Hongjing; Wang, Liang

2018-06-01

Design of highly active and cost-effective electrocatalysts is very important for the generation of hydrogen by electrochemical water-splitting. Herein, we report the fabrication of ultrathin nitrogen-doped graphitized carbon shell encapsulating CoRu bimetallic nanoparticles (CoRu@NCs) and demonstrate their promising feasibility for efficiently catalyzing the hydrogen evolution reaction (HER) over a wide pH range. The resultant CoRu@NC nanohybrids possess an alloy-carbon core-shell structure with encapsulated low-ruthenium-content CoRu bimetallic alloy nanoparticles (10-30 nm) as the core and ultrathin nitrogen-doped graphitized carbon layers (2-6 layers) as the shell. Remarkably, the optimized catalyst (CoRu@NC-2 sample) with a Ru content as low as 2.04 wt% shows superior catalytic activity and excellent durability for HER in acidic, neutral, and alkaline conditions. This work offers a new method for the design and synthesis of non-platium-based electrocatalysts for HER in all-pH.

Confined Transformation Derived Ultrathin Titanate Nanosheets/ Graphene Films for Excellent Na/K Ion Storage.

PubMed

Zeng, Cheng; Xie, Fangxi; Yang, Xianfeng; Jaroniec, Mietek; Zhang, Lei; Qiao, Shizhang

2018-05-02

Confined transformation of assembled two-dimensional MXene (titanium carbide) and reduced graphene oxide (rGO) nanosheets was employed to prepare the free-standing films of the integrated ultrathin sodium titanate (NTO)/potassium titanate (KTO) nanosheets sandwiched between graphene layers. The ultrathin Ti-based nanosheets reduce the diffusion distance while rGO layers enhance conductivity. Incorporation of graphene into the titanate films produced efficient binder-free anodes for ion storage. The resulting NTO/rGO electrode for sodium ion batteries exhibited an excellent rate performance and long cycling stability characterized by reversible capacity of 72 mA h g-1 at 5 A g-1 after 10000 cycles. Moreover, flexible KTO/rGO electrode for potassium ion batteries maintained a reversible capacity of 75 mA h g-1 after 700 cycles at 2 A g-1. These results demonstrate the superiority of the unique sandwich-type electrodes. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ultrathin nitrogen-doped graphitized carbon shell encapsulating CoRu bimetallic nanoparticles for enhanced electrocatalytic hydrogen evolution

NASA Astrophysics Data System (ADS)

Xu, You; Li, Yinghao; Yin, Shuli; Yu, Hongjie; Xue, Hairong; Li, Xiaonian; Wang, Hongjing; Wang, Liang

2018-06-01

Design of highly active and cost-effective electrocatalysts is very important for the generation of hydrogen by electrochemical water-splitting. Herein, we report the fabrication of ultrathin nitrogen-doped graphitized carbon shell encapsulating CoRu bimetallic nanoparticles (CoRu@NCs) and demonstrate their promising feasibility for efficiently catalyzing the hydrogen evolution reaction (HER) over a wide pH range. The resultant CoRu@NC nanohybrids possess an alloy–carbon core–shell structure with encapsulated low-ruthenium-content CoRu bimetallic alloy nanoparticles (10–30 nm) as the core and ultrathin nitrogen-doped graphitized carbon layers (2–6 layers) as the shell. Remarkably, the optimized catalyst (CoRu@NC-2 sample) with a Ru content as low as 2.04 wt% shows superior catalytic activity and excellent durability for HER in acidic, neutral, and alkaline conditions. This work offers a new method for the design and synthesis of non-platium-based electrocatalysts for HER in all-pH.

Enrichment of viable bacteria in a micro-volume by free-flow electrophoresis.

PubMed

Podszun, Susann; Vulto, Paul; Heinz, Helene; Hakenberg, Sydney; Hermann, Carsten; Hankemeier, Thomas; Urban, Gerald A

2012-02-07

Macro- to micro-volume concentration of viable bacteria is performed in a microfluidic chip. The enrichment principle is based on free flow electrophoresis and is demonstrated for Gram positive bacteria. Bacteria from a suspension flow are trapped on a gel interface that separates the trapping location from integrated actuation electrodes in order to enable non-destructive trapping. The microfluidic chip contains integrated electrolytic gas expulsion structures and phaseguides for gel and liquid handling. Trapping efficiency is systematically optimized to reach 25 times the initial concentration from a theoretical maximum of 30. Finally, enrichment from analytically relevant concentrations down to 3 × 10(2) colony forming units per millilitre is demonstrated with a trapping efficiency of 80% which represents the most important parameter in enrichment.

Detection of mechanically recovered chicken meat using capillary gel electrophoresis.

PubMed

Day, L; Brown, H

2001-05-01

This study investigated the use of capillary gel electrophoresis (CGE) as a method for differentiating between raw mechanically recovered chicken meat (MRM) and hand deboned chicken breast meat (HDM). Twenty samples of MRM were obtained and twenty samples of HDM were prepared in the laboratory. They were extracted and analysed using Prosort™ SDS-protein analysis reagent. There were obvious differences in the relative peak areas within the profiles obtained which distinguished raw MRM from raw HDM; specifically, that of haemoglobin was higher in MRM. Using the peak area of haemoglobin and its ratio to other peaks, the technique was tested using composite MRM-HDM mixtures. The results suggest that it is possible to differentiate mixtures containing 7.5% MRM from that of 0% MRM using the CGE method.

Quantitation of yeast total proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for uniform loading.

PubMed

Sheen, Hyukho

2016-04-01

Proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer are difficult to quantitate due to SDS and reducing agents being in the buffer. Although acetone precipitation has long been used to clean up proteins from detergents and salts, previous studies showed that protein recovery from acetone precipitation varies from 50 to 100% depending on the samples tested. Here, this article shows that acetone precipitates proteins highly efficiently from SDS-PAGE sample buffer and that quantitative recovery is achieved in 5 min at room temperature. Moreover, precipitated proteins are resolubilized with urea/guanidine, rather than with SDS. Thus, the resolubilized samples are readily quantifiable with Bradford reagent without using SDS-compatible assays. Copyright © 2016 Elsevier Inc. All rights reserved.

Detection and Identification of Gastrointestinal Lactobacillus Species by Using Denaturing Gradient Gel Electrophoresis and Species-Specific PCR Primers

PubMed Central

Walter, J.; Tannock, G. W.; Tilsala-Timisjarvi, A.; Rodtong, S.; Loach, D. M.; Munro, K.; Alatossava, T.

2000-01-01

Denaturing gradient gel electrophoresis (DGGE) of DNA fragments obtained by PCR amplification of the V2-V3 region of the 16S rRNA gene was used to detect the presence of Lactobacillus species in the stomach contents of mice. Lactobacillus isolates cultured from human and porcine gastrointestinal samples were identified to the species level by using a combination of DGGE and species-specific PCR primers that targeted 16S-23S rRNA intergenic spacer region or 16S rRNA gene sequences. The identifications obtained by this approach were confirmed by sequencing the V2-V3 region of the 16S rRNA gene and by a BLAST search of the GenBank database. PMID:10618239

Multilocus Sequence Typing Has Better Discriminatory Ability for Typing Vibrio cholerae than Does Pulsed-Field Gel Electrophoresis and Provides a Measure of Phylogenetic Relatedness

PubMed Central

Kotetishvili, Mamuka; Stine, O. Colin; Chen, Yuansha; Kreger, Arnold; Sulakvelidze, Alexander; Sozhamannan, Shanmuga; Morris, Jr., J. Glenn

2003-01-01

Twenty-two Vibrio cholerae isolates, including some from “epidemic” (O1 and O139) and “nonepidemic” serogroups, were characterized by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) by using three housekeeping genes, gyrB, pgm, and recA; sequence data were also obtained for the virulence-associated genes tcpA, ctxA, and ctxB. Even with the small number of loci used, MLST had better discriminatory ability than did PFGE. On MLST analysis, there was clear clustering of epidemic serogroups; much greater diversity was seen among tcpA- and ctxAB-positive V. cholerae strains from other, nonepidemic serogroups, with a number of tcpA and ctxAB alleles identified. PMID:12734277

Aeromonas isolates from human diarrheic stool and groundwater compared by pulsed-field gel electrophoresis.

PubMed

Borchardt, Mark A; Stemper, Mary E; Standridge, Jon H

2003-02-01

Gastrointestinal infections of Aeromonas species are generally considered waterborne; for this reason, Aeromonas hydrophila has been placed on the United States Environmental Protection Agency Contaminant Candidate List of emerging pathogens in drinking water. In this study, we compared pulsed-field gel electrophoresis patterns of Aeromonas isolates from stool specimens of patients with diarrhea with Aeromonas isolates from patients' drinking water. Among 2,565 diarrheic stool specimens submitted to a Wisconsin clinical reference laboratory, 17 (0.66%) tested positive for Aeromonas. Groundwater isolates of Aeromonas were obtained from private wells throughout Wisconsin and the drinking water of Aeromonas-positive patients. The analysis showed that the stool and drinking water isolates were genetically unrelated, suggesting that in this population Aeromonas gastrointestinal infections were not linked with groundwater exposures.

In vitro study of prebiotic properties of levan-type exopolysaccharides from Lactobacilli and non-digestible carbohydrates using denaturing gradient gel electrophoresis.

PubMed

Bello, F D; Walter, J; Hertel, C; Hammes, W P

2001-07-01

Batch cultures inoculated with human faeces were used to study the prebiotic properties of levan-type exopolysaccharides (EPS) from Lactobacillus sanfranciscensis as well as levan, inulin, and fructooligosaccharide (FOS). Denaturing gradient gel electrophoresis of 16S rDNA fragments generated by PCR with universal primers was used to analyse the cultures. Characteristic changes were revealed in the composition of the gut bacteria during fermentation of the carbohydrates. An enrichment of Bifidobacterium spp. was found for the EPS and inulin but not for levan and FOS. The bifidogenic effect of the EPS was confirmed by culturing on selective medium. In addition, the use of EPS and FOS resulted in enhanced growth of Eubacterium biforme and Clostridium perfringens, respectively.

Aeromonas Isolates from Human Diarrheic Stool and Groundwater Compared by Pulsed-Field Gel Electrophoresis

PubMed Central

Stemper, Mary E.; Standridge, Jon H.

2003-01-01

Gastrointestinal infections of Aeromonas species are generally considered waterborne; for this reason, Aeromonas hydrophila has been placed on the United States Environmental Protection Agency Contaminant Candidate List of emerging pathogens in drinking water. In this study, we compared pulsed-field gel electrophoresis patterns of Aeromonas isolates from stool specimens of patients with diarrhea with Aeromonas isolates from patients’ drinking water. Among 2,565 diarrheic stool specimens submitted to a Wisconsin clinical reference laboratory, 17 (0.66%) tested positive for Aeromonas. Groundwater isolates of Aeromonas were obtained from private wells throughout Wisconsin and the drinking water of Aeromonas-positive patients. The analysis showed that the stool and drinking water isolates were genetically unrelated, suggesting that in this population Aeromonas gastrointestinal infections were not linked with groundwater exposures. PMID:12603994

Identification of a haptoglobin-hemoglobin complex in the Alaskan Least Cisco (Coregonus sardinella).

PubMed

Wahl, S M; Boger, J K; Michael, V; Duffy, L K

1992-01-01

The hemoglobin and a hemoglobin binding protein have been characterized in the Arctic fish (Coregonus sardinella). The evolutionary significance of the hemoglobin and plasma protein differences between fish and mammals is still unresolved. Blood samples from the Alaskan Least Cisco were separated into plasma and hemoglobin fractions and the proteins in these fractions were analyzed both by alkaline agarose gel electrophoresis, by isolelectric focusing, and by capillary electrophoresis. Staining the plasma proteins gels with o-dianisidine revealed hemoglobin containing protein complexes. A hemoglobin-containing band was observed in hemolyzed plasma which did not migrate with free hemoglobin, and is believed to be hemoglobin-haptoglobin complex. Size exclusion chromatography further characterized the hemoglobin as disassociating freely into dimers, and hemoglobin-haptoglobin complex having a molecular weight greater then 200,000 daltons.

Determining thickness and refractive index from free-standing ultra-thin polymer films with spectroscopic ellipsometry

DOE PAGES

Hilfiker, James N.; Stadermann, Michael; Sun, Jianing; ...

2016-08-27

It is a well-known challenge to determine refractive index (n) from ultra-thin films where the thickness is less than about 10 nm. In this paper, we discovered an interesting exception to this issue while characterizing spectroscopic ellipsometry (SE) data from isotropic, free-standing polymer films. Ellipsometry analysis shows that both thickness and refractive index can be independently determined for free-standing films as thin as 5 nm. Simulations further confirm an orthogonal separation between thickness and index effects on the experimental SE data. Effects of angle of incidence and wavelength on the data and sensitivity are discussed. Finally, while others have demonstratedmore » methods to determine refractive index from ultra-thin films, our analysis provides the first results to demonstrate high-sensitivity to the refractive index from ultra-thin layers.« less

Fabrication of Inverted Bulk-Heterojunction Organic Solar Cell with Ultrathin Titanium Oxide Nanosheet as an Electron-Extracting Buffer Layer

NASA Astrophysics Data System (ADS)

Itoh, Eiji; Maruyama, Yasutake; Fukuda, Katsutoshi

2012-02-01

The contributions and deposition conditions of ultrathin titania nanosheet (TN) crystallites were studied in an inverted bulk-heterojunction (BHJ) cell in indium tin oxide (ITO)/titania nanosheet/poly(3-hexylthiophene) (P3HT):phenyl-C61-butyric acid methylester (PCBM) active layer/MoOx/Ag multilayered photovoltaic devices. Only one or two layers of poly(diallyldimethylammonium chloride) (PDDA) and TN multilayered film deposited by the layer-by-layer deposition technique effectively decreased the leakage current and increased both open circuit voltage (VOC) and fill factor (FF), and power conversion efficiency (η) was increased nearly twofold by the insertion of two TN layers. The deposition of additional TN layers caused the reduction in FF, and the abnormal S-shaped curves above VOC for the devices with three and four TN layers were ascribed to the interfacial potential barrier at the ITO/TN interface and the series resistance across the multilayers of TN and PDDA. The performance of the BHJ cell with TN was markedly improved, and the S-shaped curves were eliminated following the the insertion of anatase-phase titanium dioxide between the ITO and TN layers owing to the decrease in the interfacial potential barrier.

Fabrication and characterization of ultrathin dextran layers: Time dependent nanostructure in aqueous environments revealed by OWLS.

PubMed

Saftics, Andras; Kurunczi, Sándor; Szekrényes, Zsolt; Kamarás, Katalin; Khánh, Nguyen Quoc; Sulyok, Attila; Bősze, Szilvia; Horvath, Robert

2016-10-01

Surface coatings of the polysaccharide dextran and its derivatives are key ingredients especially in label-free biosensors for the suppression of non-specific binding and for receptor immobilization. Nevertheless, the nanostructure of these ultrathin coatings and its tailoring by the variation of the preparation conditions have not been profoundly characterized and understood. In this work carboxymethylated dextran (CMD) was prepared and used for fabricating ultrathin surface coatings. A grafting method based on covalent coupling to aminosilane- and epoxysilane-functionalized surfaces was applied to obtain thin CMD layers. The carboxyl moiety of the CMD was coupled to the aminated surface by EDC-NHS reagents, while CMD coupling through epoxysilane molecules was performed without any additional reagents. The surface analysis following the grafting procedures consisted of X-ray photoelectron spectroscopy (XPS), attenuated total reflection infrared spectroscopy (ATR-IR), spectroscopic ellipsometry, atomic force microscopy (AFM) and optical waveguide lightmode spectroscopy (OWLS). The XPS and AFM measurements showed that the grafting resulted in a very thin dextran layer of a few nanometers. The OWLS method allowed devising the structure of the interfacial dextran layers by the evaluation of the optogeometrical parameters. The alteration in the nanostructure of the CMD layer with the chemical composition of the silane coverage and the pH of the grafting solution was revealed by in situ OWLS, specifically, lain down chains were found to be prevalent on the surface under neutral and basic conditions on epoxysilylated surfaces. The developed methodologies allowed to design and fabricate nanometer scale CMD layers with well-controlled surface structure, which are very difficult to characterize in aqueous environments using present instrumentations and highly hydrated surface layers. Copyright © 2016 Elsevier B.V. All rights reserved.

From trace evidence to bioinformatics: putting bryophytes into molecular biology education.

PubMed

Fuselier, Linda; Bougary, Azhar; Malott, Michelle

2011-01-01

Students benefit most from their science education when they participate fully in the process of science in the context of real-world problems. We describe a student-directed open-inquiry lab experience that has no predetermined outcomes and requires students to engage in all components of scientific inquiry from posing a question through evaluating and reporting results. Over 5 weeks, students learn how bryophytes are used in forensics and become proficient in important molecular biology lab skills including DNA isolation, polymerase chain reaction, gel electrophoresis, capillary electrophoresis, and genotyping. For this portion of the experience, there is no specialized equipment necessary outside of gel electrophoresis supplies and a thermocycler. In an optional extension of the experience, students sequence a plastid intron and use introductory bioinformatics skills to identify species related to their forensics case. Students who participated in the lab experience performed well on content-based assessment, and student attitudes toward the experience were positive and indicative of engaged learning. The lab experience is easily modified for higher or lower level courses and can be used in secondary education. Copyright © 2011 Wiley Periodicals, Inc.

Carbon Nanotube Networks as Nanoscaffolds for Fabricating Ultrathin Carbon Molecular Sieve Membranes.

PubMed

Hou, Jue; Zhang, Huacheng; Hu, Yaoxin; Li, Xingya; Chen, Xiaofang; Kim, Seungju; Wang, Yuqi; Simon, George P; Wang, Huanting

2018-06-13

Carbon molecular sieve (CMS) membranes have shown great potential for gas separation owing to their low cost, good chemical stability, and high selectivity. However, most of the conventional CMS membranes exhibit low gas permeance due to their thick active layer, which limits their practical applications. Herein, we report a new strategy for fabricating CMS membranes with a 100 nm-thick ultrathin active layer using poly(furfuryl alcohol) (PFA) as a carbon precursor and carbon nanotubes (CNTs) as nanoscaffolds. CNT networks are deposited on a porous substrate as nanoscaffolds, which guide PFA solution to effectively spread over the substrate and form a continuous layer, minimizing the penetration of PFA into the pores of the substrate. After pyrolysis process, the CMS membranes with 100-1000 nm-thick active layer can be obtained by adjusting the CNT loading. The 322 nm-thick CMS membrane exhibits the best trade-off between the gas permeance and selectivity, a H 2 permeance of 4.55 × 10 -8 mol m -2 s -1 Pa -1 , an O 2 permeance of 2.1 × 10 -9 mol m -2 s -1 Pa -1 , and an O 2 /N 2 ideal selectivity of 10.5, which indicates the high quality of the membrane produced by this method. This work provides a simple, efficient strategy for fabricating ultrathin CMS membranes with high selectivity and improved gas flux.

Naturally formed ultrathin V2O5 heteroepitaxial layer on VO2/sapphire(001) film

NASA Astrophysics Data System (ADS)

Littlejohn, Aaron J.; Yang, Yunbo; Lu, Zonghuan; Shin, Eunsung; Pan, KuanChang; Subramanyam, Guru; Vasilyev, Vladimir; Leedy, Kevin; Quach, Tony; Lu, Toh-Ming; Wang, Gwo-Ching

2017-10-01

Vanadium dioxide (VO2) and vanadium pentoxide (V2O5) thin films change their properties in response to external stimuli such as photons, temperature, electric field and magnetic field and have applications in electronics, optical devices, and sensors. Due to the multiple valence states of V and non-stoichiometry in thin films, it is challenging to grow epitaxial, single-phase V-oxide on a substrate, or a heterostructure of two epitaxial V-oxides. We report the formation of a heterostructure consisting of a few nm thick ultrathin V2O5 epitaxial layer on pulsed laser deposited tens of nm thick epitaxial VO2 thin films grown on single crystal Al2O3(001) substrates without post annealing of the VO2 film. The simultaneous observation of the ultrathin epitaxial V2O5 layer and VO2 epitaxial film is only possible by our unique reflection high energy electron diffraction pole figure analysis. The out-of-plane and in-plane epitaxial relationships are V2O5[100]||VO2[010]||Al2O3[001] and V2O5[03 2 bar ]||VO2[100]||Al2O3[1 1 bar 0], respectively. The existence of the V2O5 layer on the surface of the VO2 film is also supported by X-ray photoelectron spectroscopy and Raman spectroscopy.

'One-component' ultrathin multilayer films based on poly(vinyl alcohol) as stabilizing coating for phenytoin-loaded liposomes.

PubMed

Zasada, Katarzyna; Łukasiewicz-Atanasov, Magdalena; Kłysik, Katarzyna; Lewandowska-Łańcucka, Joanna; Gzyl-Malcher, Barbara; Puciul-Malinowska, Agnieszka; Karewicz, Anna; Nowakowska, Maria

2015-11-01

Ultrathin "one-component" multilayer polymeric films for potential biomedical applications were designed based on polyvinyl alcohol,-a non-toxic, fully degradable synthetic polymer. Good uniformity of the obtained film and adequate adsorption properties of the polymeric layers were achieved by functional modification of the polymer, which involved synthesis of cationic and anionic derivatives. Synthesized polymers were characterized by FTIR, NMR spectroscopy, dynamic light scattering measurements and elemental analysis. The layer by layer assembly technique was used to build up a multilayer film and this process was followed using UV-Vis spectroscopy and ellipsometry. The morphology and thickness of the obtained multilayered film material was evaluated by atomic force microscopy (AFM). Preliminary studies on the application of the obtained multilayer film for coating of liposomal nanocarriers containing phenytoin, an antiarrhythmic drug, were performed. The coating effectively stabilizes liposomes and the effect increases with an increasing number of deposited layers until the polymeric film reaches the optimal thickness. The obtained release profiles suggest that bilayer-coated liposomes release phenytoin less rapidly than uncoated ones. The cytotoxicity studies performed for all obtained nanocarriers confirmed that none of them has negative effect on cell viability. All of the performed experiments suggest that liposomes coated with ultrathin film obtained from PVA derivatives can be attractive drug nanocarriers. Copyright © 2015 Elsevier B.V. All rights reserved.

New ultrathin film heterostructure for low-e application by sputtering technique: a theoretical and experimental study

NASA Astrophysics Data System (ADS)

Ruíz-Robles, M. A.; Abundiz-Cisneros, N.; Bender-Pérez, C. E.; Gutiérrez-Lazos, C. D.; Fundora-Cruz, A.; Solís-Pomar, F.; Pérez-Tijerina, E.

2018-03-01

The design and optical characterization by UV–vis transmittance of ultrathin low-emissivity (low-e) windows by reactive sputtering are reported. Two heterostructures on a glass substrate were considered for the low-e windows. The first heterostructure is Si3N4/TiO2/ZnO/Ag/SnO2/Si3N4 and the second is Si3N4/Ag/Si3N4. The transmittance and reflectance of these heterostructures were simulated to determine the required thickness of each layer. The first heterostructure exhibited maximum transmittance of 85% at 550 nm, slightly higher than the one determined by simulation and less than 50% transmittance in the near-infrared region (900 nm). The second heterostructure exhibited transmittance greater than 86% at 550 nm and <50% transmittance in the near-infrared region. In addition, we found that the bandwidth and maximum position of the transmittance depend on the Si3N4 layer thickness. Specifically, the thickness of the first Si3N4 layer allows the modulation of the transmittance bandwidth and the thickness of the second Si3N4 layer allows the modulation of the maximum position. The low-e windows were protected by the deposition of an ultrathin film of NiCr alloy (Ni 80%, Cr 20%) that preserved the optical characteristics and decreased the maximum of the transmittance only by 3%.

Extremely Vivid, Highly Transparent, and Ultrathin Quantum Dot Light-Emitting Diodes.

PubMed

Choi, Moon Kee; Yang, Jiwoong; Kim, Dong Chan; Dai, Zhaohe; Kim, Junhee; Seung, Hyojin; Kale, Vinayak S; Sung, Sae Jin; Park, Chong Rae; Lu, Nanshu; Hyeon, Taeghwan; Kim, Dae-Hyeong

2018-01-01

Displaying information on transparent screens offers new opportunities in next-generation electronics, such as augmented reality devices, smart surgical glasses, and smart windows. Outstanding luminance and transparency are essential for such "see-through" displays to show vivid images over clear background view. Here transparent quantum dot light-emitting diodes (Tr-QLEDs) are reported with high brightness (bottom: ≈43 000 cd m -2 , top: ≈30 000 cd m -2 , total: ≈73 000 cd m -2 at 9 V), excellent transmittance (90% at 550 nm, 84% over visible range), and an ultrathin form factor (≈2.7 µm thickness). These superb characteristics are accomplished by novel electron transport layers (ETLs) and engineered quantum dots (QDs). The ETLs, ZnO nanoparticle assemblies with ultrathin alumina overlayers, dramatically enhance durability of active layers, and balance electron/hole injection into QDs, which prevents nonradiative recombination processes. In addition, the QD structure is further optimized to fully exploit the device architecture. The ultrathin nature of Tr-QLEDs allows their conformal integration on various shaped objects. Finally, the high resolution patterning of red, green, and blue Tr-QLEDs (513 pixels in. -1 ) shows the potential of the full-color transparent display. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Spot quantification in two dimensional gel electrophoresis image analysis: comparison of different approaches and presentation of a novel compound fitting algorithm

PubMed Central

2014-01-01

Background Various computer-based methods exist for the detection and quantification of protein spots in two dimensional gel electrophoresis images. Area-based methods are commonly used for spot quantification: an area is assigned to each spot and the sum of the pixel intensities in that area, the so-called volume, is used a measure for spot signal. Other methods use the optical density, i.e. the intensity of the most intense pixel of a spot, or calculate the volume from the parameters of a fitted function. Results In this study we compare the performance of different spot quantification methods using synthetic and real data. We propose a ready-to-use algorithm for spot detection and quantification that uses fitting of two dimensional Gaussian function curves for the extraction of data from two dimensional gel electrophoresis (2-DE) images. The algorithm implements fitting using logical compounds and is computationally efficient. The applicability of the compound fitting algorithm was evaluated for various simulated data and compared with other quantification approaches. We provide evidence that even if an incorrect bell-shaped function is used, the fitting method is superior to other approaches, especially when spots overlap. Finally, we validated the method with experimental data of urea-based 2-DE of Aβ peptides andre-analyzed published data sets. Our methods showed higher precision and accuracy than other approaches when applied to exposure time series and standard gels. Conclusion Compound fitting as a quantification method for 2-DE spots shows several advantages over other approaches and could be combined with various spot detection methods. The algorithm was scripted in MATLAB (Mathworks) and is available as a supplemental file. PMID:24915860

Modified Terminal Restriction Fragment Analysis for Quantifying Telomere Length Using In-gel Hybridization.

PubMed

Jenkins, Frank J; Kerr, Charles M; Fouquerel, Elise; Bovbjerg, Dana H; Opresko, Patricia L

2017-07-10

There are several different techniques for measuring telomere length, each with their own advantages and disadvantages. The traditional approach, Telomere Restriction Fragment (TRF) analysis, utilizes a DNA hybridization technique whereby genomic DNA samples are digested with restriction enzymes, leaving behind telomere DNA repeats and some sub-telomeric DNA. These are separated by agarose gel electrophoresis, transferred to a filter membrane and hybridized to oligonucleotide probes tagged with either chemiluminescence or radioactivity to visualize telomere restriction fragments. This approach, while requiring a larger quantity of DNA than other techniques such as PCR, can measure the telomere length distribution of a population of cells and allows measurement expressed in absolute kilobases. This manuscript demonstrates a modified DNA hybridization procedure for determining telomere length. Genomic DNA is first digested with restriction enzymes (that do not cut telomeres) and separated by agarose gel electrophoresis. The gel is then dried and the DNA is denatured and hybridized in situ to a radiolabeled oligonucleotide probe. This in situ hybridization avoids loss of telomere DNA and improves signal intensity. Following hybridization, the gels are imaged utilizing phosphor screens and the telomere length is quantified using a graphing program. This procedure was developed by the laboratories of Drs. Woodring Wright and Jerry Shay at the University of Texas Southwestern 1 , 2 . Here, we present a detailed description of this procedure, with some modifications.

Ultrathin Nickel Hydroxide and Oxide Nanosheets: Synthesis, Characterizations and Excellent Supercapacitor Performances

PubMed Central

Zhu, Youqi; Cao, Chuanbao; Tao, Shi; Chu, Wangsheng; Wu, Ziyu; Li, Yadong

2014-01-01

High-quality ultrathin two-dimensional nanosheets of α-Ni(OH)2 are synthesized at large scale via microwave-assisted liquid-phase growth under low-temperature atmospheric conditions. After heat treatment, non-layered NiO nanosheets are obtained while maintaining their original frame structure. The well-defined and freestanding nanosheets exhibit a micron-sized planar area and ultrathin thickness (<2 nm), suggesting an ultrahigh surface atom ratio with unique surface and electronic structure. The ultrathin 2D nanostructure can make most atoms exposed outside with high activity thus facilitate the surface-dependent electrochemical reaction processes. The ultrathin α-Ni(OH)2 and NiO nanosheets exhibit enhanced supercapacitor performances. Particularly, the α-Ni(OH)2 nanosheets exhibit a maximum specific capacitance of 4172.5 F g−1 at a current density of 1 A g−1. Even at higher rate of 16 A g−1, the specific capacitance is still maintained at 2680 F g−1 with 98.5% retention after 2000 cycles. Even more important, we develop a facile and scalable method to produce high-quality ultrathin transition metal hydroxide and oxide nanosheets and make a possibility in commercial applications. PMID:25168127

Microfluidic device having an immobilized pH gradient and PAGE gels for protein separation and analysis

DOEpatents

Sommer, Gregory J.; Hatch, Anson V.; Singh, Anup K.; Wang, Ying-Chih

2012-12-11

Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

Microfluidic device having an immobilized pH gradient and page gels for protein separation and analysis

DOEpatents

Sommer, Gregory J; Hatch, Anson V; Singh, Anup K; Wang, Ying-Chih

2014-05-20

Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

Data on DNA gel sample load, gel electrophoresis, PCR and cost analysis.

PubMed

Kuhn, Ramona; Böllmann, Jörg; Krahl, Kathrin; Bryant, Isaac Mbir; Martienssen, Marion

2018-02-01

The data presented in this article provide supporting information to the related research article "Comparison of ten different DNA extraction procedures with respect to their suitability for environmental samples" (Kuhn et al., 2017) [1]. In that article, we compared the suitability of ten selected DNA extraction methods based on DNA quality, purity, quantity and applicability to universal PCR. Here we provide the data on the specific DNA gel sample load, all unreported gel images of crude DNA and PCR results, and the complete cost analysis for all tested extraction procedures and in addition two commercial DNA extraction kits for soil and water.

Bioorthogonal layer-by-layer encapsulation of pancreatic islets via hyperbranched polymers

PubMed Central

Gattás-Asfura, Kerim M.; Stabler, Cherie L.

2013-01-01

The encapsulation of viable tissues via layer-by-layer polymer assembly provides a versatile platform for cell surface engineering, with nanoscale control over capsule properties. Herein, we report the development of a hyperbranched polymer-based, ultrathin capsule architecture expressing bioorthogonal functionality and tailored physiochemical properties. Random carbodiimide-based condensation of 3,5-dicarboxyphenyl glycineamide on alginate yielded a highly branched polysaccharide with multiple, spatially restricted, and readily functionalizable terminal carboxylate moieties. Poly(ethylene glycol) (PEG) was utilized to link azido end groups to the structured alginate. Together with phosphine functionalized poly(amido amine) (PAMAM) dendrimer, nanoscale layer-by-layer coatings, covalently stabilized via Staudinger ligation, were assembled onto solid surfaces and pancreatic islets. The effects of electrostatic and/or bioorthogonal covalent interlayer interactions on the resulting coating efficiency and stability, as well as pancreatic islet viability and function, were studied. These hyperbranched polymers provide a flexible platform for the formation of covalently stabilized ultrathin coatings on viable cells and tissues. In addition, the hyperbranched nature of the polymers presents a highly functionalized surface capable of bioorthogonal conjugation of additional bioactive or labeling motifs. PMID:24063764

Evaluation and optimisation of preparative semi-automated electrophoresis systems for Illumina library preparation.

PubMed

Quail, Michael A; Gu, Yong; Swerdlow, Harold; Mayho, Matthew

2012-12-01

Size selection can be a critical step in preparation of next-generation sequencing libraries. Traditional methods employing gel electrophoresis lack reproducibility, are labour intensive, do not scale well and employ hazardous interchelating dyes. In a high-throughput setting, solid-phase reversible immobilisation beads are commonly used for size-selection, but result in quite a broad fragment size range. We have evaluated and optimised the use of two semi-automated preparative DNA electrophoresis systems, the Caliper Labchip XT and the Sage Science Pippin Prep, for size selection of Illumina sequencing libraries. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sol-gel layers for ceramic microsystems application

NASA Astrophysics Data System (ADS)

Czok, Mateusz; Golonka, Leszek

2016-11-01

This paper describes research on sol-gel solutions preparation process. Utilize of a sol-gel layers in the LTCC technology for reduction of surface roughness and influence on the ceramics properties is examined and described. The influence of sol-gel layer on possible sedimentation of dyes or biological substances in channels, mixers or chambers of ceramic microfluidic structures was investigated. Moreover, properties of sol-gel coated surfaces have been precisely examined and described. Finally, positive results of conducted experiments made it possible to design and manufacture a simple microfluidic ceramic structure, with embedded protective layer of sol-gel, for fluorescence measurements.

Structural and electrical characterization of ultra-thin SrTiO3 tunnel barriers grown over YBa2Cu3O7 electrodes for the development of high Tc Josephson junctions.

PubMed

Félix, L Avilés; Sirena, M; Guzmán, L A Agüero; Sutter, J González; Vargas, S Pons; Steren, L B; Bernard, R; Trastoy, J; Villegas, J E; Briático, J; Bergeal, N; Lesueur, J; Faini, G

2012-12-14

The transport properties of ultra-thin SrTiO(3) (STO) layers grown over YBa(2)Cu(3)O(7) electrodes were studied by conductive atomic force microscopy at the nano-scale. A very good control of the barrier thickness was achieved during the deposition process. A phenomenological approach was used to obtain critical parameters regarding the structural and electrical properties of the system. The STO layers present an energy barrier of 0.9 eV and an attenuation length of 0.23 nm, indicating very good insulating properties for the development of high-quality Josephson junctions.

A new bimetallic plasmonic photocatalyst consisting of gold(core)-copper(shell) nanoparticle and titanium(IV) oxide support

NASA Astrophysics Data System (ADS)

Sato, Yuichi; Naya, Shin-ichi; Tada, Hiroaki

2015-10-01

Ultrathin Cu layers (˜2 atomic layers) have been selectively formed on the Au surfaces of Au nanoparticle-loaded rutile TiO2 (Au@Cu/TiO2) by a deposition precipitation-photodeposition technique. Cyclic voltammetry and photochronopotentiometry measurements indicate that the reaction proceeds via the underpotential deposition. The ultrathin Cu shell drastically increases the activity of Au/TiO2 for the selective oxidation of amines to the corresponding aldehydes under visible-light irradiation (λ > 430 nm). Photochronoamperometry measurements strongly suggest that the striking Cu shell effect stems from the enhancement of the charge separation in the localized surface plasmon resonance-excited Au/TiO2.

How-to-Do-It: Biotechnology in Three Days.

ERIC Educational Resources Information Center

Gardner, Alan M.

1988-01-01

Outlines a three-day unit for presenting biotechnology. States that the approach surveys the processes of enzyme restriction, ligation, transformations of recombinant plasmids, and gel electrophoresis. Diagrams accompany the article. (RT)

Effects of heat/citric acid reprocessing on high-flux polysulfone dialyzers.

PubMed

Cornelius, Rena M; McClung, W Glenn; Richardson, Robert M A; Estridge, Charles; Plaskos, Nicholas; Yip, Christopher M; Brash, John L

2002-01-01

The surface features, morphology, and tensile properties of fibers obtained from pristine, reprocessed, and reused Fresenius Polysulfone High-Flux (Hemoflow F80A) hemodialyzers have been studied. Scanning electron microscopy of the dialyzer fibers revealed a dense skin layer on the inner surface of the membrane and a relatively thick porous layer on the outer surface. Transmission electron microscopy and atomic force microscopy showed an alteration in membrane morphology due to reprocessing and reuse, or to a deposition of blood-borne material on the membrane that is not removed with reprocessing. Fluorescent microscopy images also showed that a fluorescent material not removed by heat/citric acid reprocessing builds up with continued use of the dialyzers. The tensile properties of the dialyzer fibers were not affected by the heat/citric acid reprocessing procedure. The protein layers formed on pristine and reused hemodialyzer membranes during clinical use were also studied using sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting. A considerable amount of protein was found on the blood side of single and multiple use dialyzers. Proteins adsorbed on the dialysate side of the membrane were predominantly in the molecular weight region below 30 kDa. Little protein was detected on the membranes of reprocessed hemodialyzers.

Purification of Lactobacillus acidophilus surface-layer protein and its immunomodulatory effects on RAW264.7 cells.

PubMed

Zhang, Dandan; Wu, Mengting; Guo, Yuxing; Xun, Mingyue; Wang, Wenwen; Wu, Zhen; Pan, Daodong

2017-09-01

Surface-layer proteins (SLP) have been found in the outermost layer of the cell wall in many types of lactobacillus are considered to be an important factor with respect to intestinal immunity. The present study compared the effects of SLP extracted by different concentrations of LiCl and carbamide, and subsequently identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism and differential scanning calorimetry. Furthermore, RAW 264.7 cells were used to evaluate the immunomodulatory effects of SLP. SLP were derived from Lactobacillus acidophilus CICC6074 with a molecular weight of 46 kDa, and consisted of 16.9% α-helix, 42.3% β-sheet, 20.8% β-turns and 22.5% random coils. SLP promoted NO secretion and higher quantities of NO were produced as the SLP concentrations increased. SLP concentrations over 50 µg mL -1 significantly decreased the amount of tumor necrosis factor-α secreted by RAW264.7 cells. SLP can trigger immunomodulatory effects in RAW 264.7 cells. This provides crucial information that will enable the further use of L. acidophilus in food, medicine and other products. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Appearance and disappearance of ferromagnetism in ultrathin LaMnO3 on SrTiO3 substrate: A viewpoint from first principles

NASA Astrophysics Data System (ADS)

An, Ming; Weng, Yakui; Zhang, Huimin; Zhang, Jun-Jie; Zhang, Yang; Dong, Shuai

2017-12-01

The intrinsic magnetic state (ferromagnetic or antiferromagnetic) of ultrathin LaMnO3 films on the most commonly used SrTiO3 substrate is a long-existing question under debate. Either strain effect or nonstoichiometry was argued to be responsible for the experimental ferromagnetism. In a recent experiment [X. R. Wang, C. J. Li, W. M. Lü, T. R. Paudel, D. P. Leusink, M. Hoek, N. Poccia, A. Vailionis, T. Venkatesan, J. M. D. Coey, E. Y. Tsymbal, Ariando, and H. Hilgenkamp, Science 349, 716 (2015), 10.1126/science.aaa5198], one more mechanism, namely, the self-doping due to polar discontinuity, was argued to be the driving force of ferromagnetism beyond the critical thickness. Here systematic first-principles calculations have been performed to check these mechanisms in ultrathin LaMnO3 films as well as superlattices. Starting from the very precise descriptions of both LaMnO3 and SrTiO3, it is found that the compressive strain is the dominant force for the appearance of ferromagnetism, while the open surface with oxygen vacancies leads to the suppression of ferromagnetism. Within LaMnO3 layers, the charge reconstructions involve many competitive factors and certainly go beyond the intuitive polar catastrophe model established for LaAlO3/SrTiO3 heterostructures. Our paper not only explains the long-term puzzle regarding the magnetism of ultrathin LaMnO3 films but also sheds light on how to overcome the notorious magnetic dead layer in ultrathin manganites.

Sample collection system for gel electrophoresis

DOEpatents

Olivares, Jose A.; Stark, Peter C.; Dunbar, John M.; Hill, Karen K.; Kuske, Cheryl R.; Roybal, Gustavo

2004-09-21

An automatic sample collection system for use with an electrophoretic slab gel system is presented. The collection system can be used with a slab gel have one or more lanes. A detector is used to detect particle bands on the slab gel within a detection zone. Such detectors may use a laser to excite fluorescently labeled particles. The fluorescent light emitted from the excited particles is transmitted to low-level light detection electronics. Upon the detection of a particle of interest within the detection zone, a syringe pump is activated, sending a stream of buffer solution across the lane of the slab gel. The buffer solution collects the sample of interest and carries it through a collection port into a sample collection vial.

Microscopic Electronic and Mechanical Properties of Ultra-Thin Layered Materials

DTIC Science & Technology

2016-07-25

Graphene single layers grown by chemical vapor deposition on single crystal Cu substrates are subject to nonuniform physisorption strains that...the observed highly nonuniform strains. 4. Connecting dopant bond type with electronic structure in N-doped graphene (reference [4]) Robust methods

Exposures of Sus scrofa to a TASER(®) conducted electrical weapon: no effects on 2-dimensional gel electrophoresis patterns of plasma proteins.

PubMed

Jauchem, James R; Cerna, Cesario Z; Lim, Tiffany Y; Seaman, Ronald L

2014-12-01

In an earlier study, we found significant changes in red-blood-cell, leukocyte, and platelet counts, and in red-blood-cell membrane proteins, following exposures of anesthetized pigs to a conducted electrical weapon. In the current study, we examined potential changes in plasma proteins [analyzed via two-dimensional gel electrophoresis (2-DGE)] following two 30 s exposures of anesthetized pigs (Sus scrofa) to a TASER (®) C2 conducted electrical weapon. Patterns of proteins, separated by 2-DGE, were consistent and reproducible between animals and between times of sampling. We determined that the blood plasma collection, handling, storage, and processing techniques we used are suitable for swine blood. There were no statistically significant changes in plasma proteins following the conducted-electrical-weapon exposures. Overall gel patterns of fibrinogen were similar to results of other studies of both pigs and humans (in control settings, not exposed to conducted electrical weapons). The lack of significant changes in plasma proteins may be added to the body of evidence regarding relative safety of TASER C2 device exposures.

Rapid detection of common Chinese glucose-6-phosphate dehydrogenase (G6PD) mutations by denaturing gradient gel electrophoresis (DGGE).

PubMed

Lam, V M; Huang, W; Lam, S T; Yeung, C Y; Johnson, P H

1996-03-01

We describe here the use of denaturing gradient gel electrophoresis (DGGE) to detect the most common Chinese glucose-6-phosphate dehydrogenase (G6PD) variants, which are the single point mutations: G-->T at nt 1376, G-->A at 1388 both in exon 12 and A-->G at nt 95 in exon 02. In each case, the mutant allele resolves well from the normal allele(s). The distinct heteroduplex bands are characteristic of a particular genotype suggesting that this feature is very useful for identifying all heterozygous carriers for this and other X-linked diseases. When the analysis is extended to other exons, DGGE scans the gene and coupled with direct sequencing, it leads to the identification of new G6PD variation(s). With this approach, we identified a mutation in exon 9 which had not been reported in Hong Kong. Since DGGE can rapidly screen many unknown samples in one gel, this approach could be used to diagnose these G6PD mutations and to identify the at-risk for counselling.

Rapid in vitro labeling procedures for two-dimensional gel fingerprinting

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

Lee, Y.F.; Fowlks, E.R.

1982-01-15

Improvements of existing in vitro procedures for labeling RNA radioactively, and modifications of the two-dimensional polyacrylamide gel electrophoresis system for making RNA fingerprints are described. These improvements are (a) inactivation of phosphatase with nitric acid at pH 2.0 eliminated the phenol-cholorform extraction step during 5'-end labeling with polynucleotide kinase and (..gamma..-/sup 32/P)ATP; (b) ZnSO/sub 4/ inactivation of R Nase T/sub 1/ results in a highly efficient procedure for 3'-end labeling with T4 ligase and (5'-/sup 32/P)pCp; and (c) a rapid 4-min procedure for variable quantity range of /sup 125/I and RNA results in a qualitative and quantitative sample for high-molecularmore » weight RNA fingerprinting. Thus, these in vitro procedures become rapid and reproducible when combined with two-dimensional gel electrophoresis which eliminates simultaneously labeled impurities. Each labeling procedure is compared, using tobacco mosaic virus, Brome mosaic virus, and polio RNA. A series of Ap-rich oligonucleotides was discovered in the inner genome of Brome mosaic Virus RNA-3.« less

New possibilities for tuning ultrathin cobalt film magnetic properties by a noble metal overlayer.

PubMed

Kisielewski, M; Maziewski, A; Tekielak, M; Wawro, A; Baczewski, L T

2002-08-19

Complementary multiscale magneto-optical studies based on the polar Kerr effect are carried out on an ultrathin cobalt wedge covered with a silver wedge and subsequently with the Au thick layer. A few monolayers of Ag are found to have a substantial effect on magnetic anisotropy, the coercivity field, and Kerr rotation. The silver overlayer thickness-driven magnetic reorientation from easy axis to easy plane generates a new type of 90 degrees magnetic wall for cobalt thicknesses between 1.3 and 1.8 nm. The tuning of the wall width in a wide range is possible. Tailoring of the overlayer structure can be used for ultrathin film magnetic patterning.

Methods for Initial Characterization of Campylobacter jejuni Bacteriophages.

PubMed

Sørensen, Martine Camilla Holst; Gencay, Yilmaz Emre; Brøndsted, Lone

2017-01-01

Here we describe an initial characterization of Campylobacter jejuni bacteriophages by host range analysis, genome size determination by pulsed-field gel electrophoresis, and receptor-type identification by screening mutants for phage sensitivity.

The Dynamics of DNA Sequencing.

ERIC Educational Resources Information Center

Morvillo, Nancy

1997-01-01

Describes a paper-and-pencil activity that helps students understand DNA sequencing and expands student understanding of DNA structure, replication, and gel electrophoresis. Appropriate for advanced biology students who are familiar with the Sanger method. (DDR)

Purification and some kinetic properties of catalase from parsley (Petroselinum hortense Hoffm., Apiaceae) leaves.

PubMed

Oztürk, Lokman; Bülbül, Metin; Elmastas, Mahfuz; Ciftçi, Mehmet

2007-01-01

In this study, catalase (CAT: EC 1.11.1.6) was purified from parsley (Petroselinum hortense) leaves; analysis of the kinetic behavior and some properties of the enzyme were investigated. The purification consisted of three steps, including preparation of homogenate, ammonium sulfate fractionation, and fractionation by DEAE-Sephadex A50 ion exchange chromatography. The enzyme was obtained with a yield of 9.5% and had a specific activity of 1126 U (mg proteins)(-1). The overall purification was about 5.83-fold. A temperature of 4 degrees C was maintained during the purification process. Enzyme activity was spectrophotometrically measured at 240 nm. In order to control the purification of the enzyme, SDS-polyacrylamide gel electrophoresis was carried out in 4% and 10% acryl amide for stacking and running gel, respectively. SDS-polyacrylamide gel electrophoresis showed a single band for the enzyme. The molecular weight was found to be 183.29 kDa by Sephadex G-200 gel filtration chromatography. The stable pH, optimum pH, and ionic strength were determined for phosphate and Tris-HCl buffer systems. In addition, K(M) and V(max) values for H(2)O(2), at optimum pH and 25 degrees C, were determined by means of Lineweaver-Burk plots.

Discriminatory usefulness of pulsed-field gel electrophoresis and sequence-based typing in Legionella outbreaks.

PubMed

Quero, Sara; García-Núñez, Marian; Párraga-Niño, Noemí; Barrabeig, Irene; Pedro-Botet, Maria L; de Simon, Mercè; Sopena, Nieves; Sabrià, Miquel

2016-06-01

To compare the discriminatory power of pulsed-field gel electrophoresis (PFGE) and sequence-based typing (SBT) in Legionella outbreaks for determining the infection source. Twenty-five investigations of Legionnaires' disease were analyzed by PFGE, SBT and Dresden monoclonal antibody. The results suggested that monoclonal antibody could reduce the number of Legionella isolates to be characterized by molecular methods. The epidemiological concordance PFGE-SBT was 100%, while the molecular concordance was 64%. Adjusted Wallace index (AW) showed that PFGE has better discriminatory power than SBT (AWSBT→PFGE = 0.767; AWPFGE→SBT = 1). The discrepancies appeared mostly in sequence type (ST) 1, a worldwide distributed ST for which PFGE discriminated different profiles. SBT discriminatory power was not sufficient verifying the infection source, especially in worldwide distributed STs, which were classified into different PFGE patterns.

[A structural protein study of the influenza A (H1N1) virus by polyacrylamide gel electrophoresis].

PubMed

Pérez Guevara, M T; Savón Valdés, C; Rivas Arjona, M; Goyenechea Hernández, A

1992-01-01

Influenza is an acute respiratory disease typically appearing as an epidemic. Three immunological types of the influenza virus are known: A, B and C. Continually, antigen changes occur, especially in type A. Therefore, a comparative study was carried out on 4 influenza A(H1N1) virus strains in relation to protein structure (surface antigens), by using polyacrylamide gel electrophoresis by the modified Laemmli method. The objective was to compare the structural proteins of the A/Havana/1292/78 (H1N1) national strain with the proteins of 3 international pattern strains. In all the cases, 6 bands were detected by densitometry. In the 4 strains studied the most abundant protein was M. Great differences between the Cuban strain and the 3 international patterns were not seen.

Protein Separation by Capillary Gel Electrophoresis: A Review

PubMed Central

Zhu, Zaifang; Lu, Joann J.; Liu, Shaorong

2011-01-01

Capillary gel electrophoresis (CGE) has been used for protein separation for more than two decades. Due to the technology advancement, current CGE methods are becoming more and more robust and reliable for protein analysis, and some of the methods have been routinely used for the analysis of protein-based pharmaceuticals and quality controls. In light of this progress, we survey 147 papers related to CGE separations of proteins and present an overview of this technology. We first introduce briefly the early development of CGE. We then review the methodology, in which we specifically describe the matrices, coatings, and detection strategies used in CGE. CGE using microfabricated channels and incorporation of CGE with two-dimensional protein separations are also discussed in this section. We finally present a few representative applications of CGE for separating proteins in real-world samples. PMID:22122927

A novel polymerase chain reaction (PCR) - denaturing gradient gel electrophoresis (DGGE) for the identification of Micrococcaceae strains involved in meat fermentations. Its application to naturally fermented Italian sausages.

PubMed

Cocolin, L; Manzano, M; Aggio, D; Cantoni, C; Comi, G

2001-05-01

A new molecular method consisting of polymerase chain reaction (PCR) amplification and denaturing gradient gel electrophoresis (DGGE) of a small fragment from the 16S rRNA gene identified the Micrococcaceae strains isolated from natural fermented Italian sausages. Lactic acid bacteria, total aerobic mesophilic flora, Enterobacteriaceae and faecal enterococci were also monitored. Micrococcaceaea control strains from international collections were used to optimise the method and 90 strains, isolated from fermented sausages, were identified by biochemical tests and PCR-DGGE. No differences were observed between the methods used. The results reported in this paper prove that Staphylococcus xylosus is the main bacterium involved in fermented sausage production, representing, from the tenth day of ripening, the only Micrococcaceaea species isolated.

High-Resolution Melting-Curve Analysis of Ligation-Mediated Real-Time PCR for Rapid Evaluation of an Epidemiological Outbreak of Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli ▿

PubMed Central

Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E.; Schön, Thomas

2011-01-01

Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE. PMID:21956981

High-resolution melting-curve analysis of ligation-mediated real-time PCR for rapid evaluation of an epidemiological outbreak of extended-spectrum-beta-lactamase-producing Escherichia coli.

PubMed

Woksepp, Hanna; Jernberg, Cecilia; Tärnberg, Maria; Ryberg, Anna; Brolund, Alma; Nordvall, Michaela; Olsson-Liljequist, Barbro; Wisell, Karin Tegmark; Monstein, Hans-Jürg; Nilsson, Lennart E; Schön, Thomas

2011-12-01

Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE.

Lupus autoantibodies target ribosomal P proteins

PubMed Central

1985-01-01

All nine SLE (systemic lupus erythematosus) sera with antiribosomal antibody activity targeted the same three ribosomal protein antigens, of molecular masses 38 and 17/19 kD when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. One serum reacted with an additional protein of approximately kD. Ribosomal subunit fractionation by composite gel electrophoresis and sucrose density ultracentrifugation showed that these proteins were part of the large subunit. Isoelectric focusing in agarose, and two-dimensional polyacrylamide gel electrophoresis revealed that the antigens had pI between 4.5 and 6.5, but that the 17/19 kD antigens were more acidic than the 38 kD antigen. Similarities in the molecular masses, charges, as well as the presence of highly conserved crossreactive epitopes, failure to bind to carboxymethylcellulose at pH 4.2, and extractability of the 17/19 kD proteins by 400 mM NH4Cl-ethanol at 0 degrees C indicated that these antigens were analogous to the proteins P0 (38 kD) and P1/P2 (17/19 kD) described previously (25, 36). Co-identity was confirmed using reference antibodies and antigen. Although antibodies to these proteins were only found in 5-10% of more than 50 sera screened by radioimmunoassay or Western blotting, the selective production of antibodies to epitopes on three (out of a total of more than 80) ribosomal proteins may provide further clues to autoantibody induction of SLE. PMID:2410526

Dynamics of ultra-thin polystyrene with and without a (artificial) dead layer studied by resonance enhanced dynamic light scattering

NASA Astrophysics Data System (ADS)

Vianna, S. D. B.; Lin, F. Y.; Plum, M. A.; Duran, H.; Steffen, W.

2017-05-01

Using non-invasive, marker-free resonance enhanced dynamic light scattering, the dynamics of capillary waves on ultrathin polystyrene films' coupling to the viscoelastic and mechanical properties have been studied. The dynamics of ultrathin polymer films is still debated. In particular the question of what influence either the solid substrate and/or the fluid-gas interface has on the dynamics and the mechanical properties of films of glass forming liquids as polymers is in the focus of the present research. As a consequence, e.g., viscosity close to interfaces and thus the average viscosity of very thin films are prone to change. This study is focused on atactic, non-entangled polystyrene thin films on the gold surface. A slow dynamic mode was observed with Vogel-Fulcher-Tammann temperature dependence, slowing down with decreasing film thickness. We tentatively attribute this relaxation mode to overdamped capillary waves because of its temperature dependence and the dispersion with a wave vector which was found. No signs of a more mobile layer at the air/polymer interface or of a "dead layer" at the solid/polymer interface were found. Therefore we investigated the influence of an artificially created dead layer on the capillary wave dynamics by introducing covalently bound polystyrene polymer brushes as anchors. The dynamics was slowed down to a degree more than expected from theoretical work on the increase of density close to the solid liquid interface—instead of a "dead layer" of 2 nm, the interaction seems to extend more than 10 nm into the polymer.

Anionic poly(p-phenylenevinylene)/layered double hydroxide ordered ultrathin films with multiple quantum well structure: a combined experimental and theoretical study.

PubMed

Yan, Dongpeng; Lu, Jun; Ma, Jing; Wei, Min; Wang, Xinrui; Evans, David G; Duan, Xue

2010-05-18

The sulfonated phenylenevinylene polyanion derivate (APPV) and exfoliated Mg-Al-layered double hydroxide (LDH) monolayers were alternatively assembled into ordered ultrathin films (UTFs) employing a layer-by-layer method, which shows uniform yellow luminescence. UV-vis absorption and fluorescence spectroscopy present a stepwise and regular growth of the UTFs upon increasing deposited cycles. X-ray diffraction, atomic force microscopy, and scanning electron microscopy demonstrate that the UTFs are orderly periodical layered structure with a thickness of 3.3-3.5 nm per bilayer. The APPV/LDH UTFs exhibit well-defined polarized photoemission characteristic with the maximum luminescence anisotropy of approximately 0.3. Moreover, the UTF exhibit longer fluorescence lifetime (3-3.85-fold) and higher photostability than the drop-casting APPV film under UV irradiation, suggesting that the existence of a LDH monolayer enhances the optical performance of the APPV polyanion. A combination study of electrochemistry and periodic density functional theory was used to investigate the electronic structure of the APPV/LDH system, illustrating that the APPV/LDH UTF is a kind of organic-inorganic hybrid multiple quantum well (MQW) structure with a low band energy of 1.7-1.8 eV, where the valence electrons of APPV can be confined into the energy wells formed by the LDH monolayers effectively. Therefore, this work not only gives a feasible method for fabricating a luminescence ultrathin film but also provides a detailed understanding of the geometric and electronic structures of photoactive polyanions confined between the LDH monolayers.

III-Nitride Digital Alloy: Electronics and Optoelectronics Properties of the InN/GaN Ultra-Short Period Superlattice Nanostructures.

PubMed

Sun, Wei; Tan, Chee-Keong; Tansu, Nelson

2017-07-27

The III-Nitride digital alloy (DA) is comprehensively studied as a short-period superlattice nanostructure consisting of ultra-thin III-Nitride epitaxial layers. By stacking the ultra-thin III-Nitride epitaxial layers periodically, these nanostructures are expected to have comparable optoelectronic properties as the conventional III-Nitride alloys. Here we carried out numerical studies on the InGaN DA showing the tunable optoelectronic properties of the III-Nitride DA. Our study shows that the energy gap of the InGaN DA can be tuned from ~0.63 eV up to ~2.4 eV, where the thicknesses and the thickness ratio of each GaN and InN ultra-thin binary layers within the DA structure are the key factors for tuning bandgap. Correspondingly, the absorption spectra of the InGaN DA yield broad wavelength tunability which is comparable to that of bulk InGaN ternary alloy. In addition, our investigation also reveals that the electron-hole wavefunction overlaps are remarkably large in the InGaN DA structure despite the existence of strain effect and build-in polarization field. Our findings point out the potential of III-Nitride DA as an artificially engineered nanostructure for optoelectronic device applications.

Ultrathin MoS2 and WS2 layers on silver nano-tips as electron emitters

NASA Astrophysics Data System (ADS)

Loh, Tamie A. J.; Tanemura, Masaki; Chua, Daniel H. C.

2016-09-01

2-dimensional (2D) inorganic analogues of graphene such as MoS2 and WS2 present interesting opportunities for field emission technology due to their high aspect ratio and good electrical conductivity. However, research on 2D MoS2 and WS2 as potential field emitters remains largely undeveloped compared to graphene. Herein, we present an approach to directly fabricate ultrathin MoS2 and WS2 onto Ag nano-tips using pulsed laser deposition at low temperatures of 450-500 °C. In addition to providing a layer of chemical and mechanical protection for the Ag nano-tips, the growth of ultrathin MoS2 and WS2 layers on Ag led to enhanced emission properties over that of pristine nano-tips due to a reduction of the effective barrier height arising from charge injection from Ag to the overlying MoS2 or WS2. For WS2 on Ag nano-tips, the phasic mixture was also an important factor influencing the field emission performance. The presence of 1T-WS2 at the metal-WS2 interface in a hybrid film of 2H/1T-WS2 leads to improvement in the field emission capabilities as compared to pure 2H-WS2 on Ag nano-tips.

Investigation of water mobility and diffusivity in hydrating micronized low-substituted hydroxypropyl cellulose, hydroxypropylmethyl cellulose, and hydroxypropyl cellulose matrix tablets by magnetic resonance imaging (MRI).

PubMed

Kojima, Masazumi; Nakagami, Hiroaki

2002-12-01

The water mobility and diffusivity in the gel-layer of hydrating low-substituted hydroxypropyl cellulose (LH41) tablets with or without a drug were investigated by magnetic resonance imaging (MRI) and compared with those properties in the gel-layer of hydroxypropylmethyl cellulose (HPMC) and hydroxypropyl cellulose (HPC) tablets. For this purpose, a localized image-analysis method was newly developed, and the spin-spin relaxation time (T(2)) and apparent self-diffusion coefficient (ADC) of water in the gel-layer were visualized in one-dimensional maps. Those maps showed that the extent of gel-layer growth in the tablets was in the order of HPC>HPMC>LH41, and there was a water mobility gradient across the gel-layers of all three tablet formulations. The T(2) and ADC in the outer parts of the gel-layers were close to those of free water. In contrast, these values in the inner parts of the gel-layer decreased progressively; suggesting that the water mobility and diffusivity around the core interface were highly restricted. Furthermore, the correlation between the T(2) of (1)H proton in the gel-layer of the tablets and the drug release rate from the tablets was observed.

Few-layered CoHPO4.3H2O ultrathin nanosheets for high performance of electrode materials for supercapacitors

NASA Astrophysics Data System (ADS)

Pang, Huan; Wang, Shaomei; Shao, Weifang; Zhao, Shanshan; Yan, Bo; Li, Xinran; Li, Sujuan; Chen, Jing; Du, Weimin

2013-06-01

Ultrathin cobalt phosphate (CoHPO4.3H2O) nanosheets are successfully synthesized by a one pot hydrothermal method. Novel CoHPO4.3H2O ultrathin nanosheets are assembled for constructing the electrodes of supercapacitors. Benefiting from the nanostructures, the as-prepared electrode shows a specific capacitance of 413 F g-1, and no obvious decay even after 3000 charge-discharge cycles. Such a quasi-two-dimensional material is a new kind of supercapacitor electrode material with high performance.Ultrathin cobalt phosphate (CoHPO4.3H2O) nanosheets are successfully synthesized by a one pot hydrothermal method. Novel CoHPO4.3H2O ultrathin nanosheets are assembled for constructing the electrodes of supercapacitors. Benefiting from the nanostructures, the as-prepared electrode shows a specific capacitance of 413 F g-1, and no obvious decay even after 3000 charge-discharge cycles. Such a quasi-two-dimensional material is a new kind of supercapacitor electrode material with high performance. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr01460f

Manipulation of Spin-Torque Generation Using Ultrathin Au

NASA Astrophysics Data System (ADS)

An, Hongyu; Haku, Satoshi; Kanno, Yusuke; Nakayama, Hiroyasu; Maki, Hideyuki; Shi, Ji; Ando, Kazuya

2018-06-01

The generation and the manipulation of current-induced spin-orbit torques are of essential interest in spintronics. However, in spite of the vital progress in spin orbitronics, electric control of the spin-torque generation still remains elusive and challenging. We report on electric control of the spin-torque generation using ionic-liquid gating of ultrathin Au. We show that by simply depositing a SiO2 capping layer on an ultrathin-Au /Ni81Fe19 bilayer, the spin-torque generation efficiency is drastically enhanced by a maximum of 7 times. This enhancement is verified to be originated from the rough ultrathin-Au /Ni81Fe19 interface induced by the SiO2 deposition, which results in the enhancement of the interface spin-orbit scattering. We further show that the spin-torque generation efficiency from the ultrathin Au film can be reversibly manipulated by a factor of 2 using the ionic gating with an external electric field within a small range of 1 V. These results pave a way towards the efficient control of the spin-torque generation in spintronic applications.

Removal of the Magnetic Dead Layer by Geometric Design

DOE PAGES

Guo, Er-jia; Roldan, Manuel; Charlton, Timothy R.; ...

2018-05-28

The proximity effect is used to engineer interface effects such as magnetoelectric coupling, exchange bias, and emergent interfacial magnetism. However, the presence of a magnetic “dead layer” adversely affects the functionality of a heterostructure. Here in this paper, it is shown that by utilizing (111) polar planes, the magnetization of a manganite ultrathin layer can be maintained throughout its thickness. Combining structural characterization, magnetometry measurements, and magnetization depth profiling with polarized neutron reflectometry, it is found that the magnetic dead layer is absent in the (111)-oriented manganite layers, however, it occurs in the films with other orientations. Quantitative analysis ofmore » local structural and elemental spatial evolutions using scanning transmission electron microscopy and electron energy loss spectroscopy reveals that atomically sharp interfaces with minimal chemical intermixing in the (111)-oriented superlattices. The polar discontinuity across the (111) interfaces inducing charge redistribution within the SrTiO 3 layers is suggested, which promotes ferromagnetism throughout the (111)-oriented ultrathin manganite layers. The approach of eliminating problematic magnetic dead layers by changing the crystallographic orientation suggests a conceptually useful recipe to engineer the intriguing physical properties of oxide interfaces, especially in low dimensionality.« less

Removal of the Magnetic Dead Layer by Geometric Design

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

Guo, Er-jia; Roldan, Manuel; Charlton, Timothy R.

The proximity effect is used to engineer interface effects such as magnetoelectric coupling, exchange bias, and emergent interfacial magnetism. However, the presence of a magnetic “dead layer” adversely affects the functionality of a heterostructure. Here in this paper, it is shown that by utilizing (111) polar planes, the magnetization of a manganite ultrathin layer can be maintained throughout its thickness. Combining structural characterization, magnetometry measurements, and magnetization depth profiling with polarized neutron reflectometry, it is found that the magnetic dead layer is absent in the (111)-oriented manganite layers, however, it occurs in the films with other orientations. Quantitative analysis ofmore » local structural and elemental spatial evolutions using scanning transmission electron microscopy and electron energy loss spectroscopy reveals that atomically sharp interfaces with minimal chemical intermixing in the (111)-oriented superlattices. The polar discontinuity across the (111) interfaces inducing charge redistribution within the SrTiO 3 layers is suggested, which promotes ferromagnetism throughout the (111)-oriented ultrathin manganite layers. The approach of eliminating problematic magnetic dead layers by changing the crystallographic orientation suggests a conceptually useful recipe to engineer the intriguing physical properties of oxide interfaces, especially in low dimensionality.« less

Effects of different wetting layers on the growth of smooth ultra-thin silver thin films

NASA Astrophysics Data System (ADS)

Ni, Chuan; Shah, Piyush; Sarangan, Andrew M.

2014-09-01

Ultrathin silver films (thickness below 10 nm) are of great interest as optical coatings on windows and plasmonic devices. However, producing these films has been a continuing challenge because of their tendency to form clusters or islands rather than smooth contiguous thin films. In this work we have studied the effect of Cu, Ge and ZnS as wetting layers (1.0 nm) to achieve ultrasmooth thin silver films. The silver films (5 nm) were grown by RF sputter deposition on silicon and glass substrates using a few monolayers of the different wetting materials. SEM imaging was used to characterize the surface properties such as island formation and roughness. Also the optical properties were measured to identify the optical impact of the different wetting layers. Finally, a multi-layer silver based structure is designed and fabricated, and its performance is evaluated. The comparison between the samples with different wetting layers show that the designs with wetting layers which have similar optical properties to silver produce the best overall performance. In the absence of a wetting layer, the measured optical spectra show a significant departure from the model predictions, which we attribute primarily to the formation of clusters.

Fabrication of Nanolaminates with Ultrathin Nanolayers Using Atomic Layer Deposition: Nucleation & Growth Issues

DTIC Science & Technology

2009-02-01

Tecnologia de Superficies y Materiales (SMCTSM), XXVII Congreso Nacional, Oaxaca, Oaxaca, Mexico, September 26, 2007. 26. "Atomic Layer Deposition of...Nanolaminates: Fabrication and Properties" (Plenary Lecture), Sociedad Mexicana de Ciencia y Tecnologia de Superficies y Materiales (SMCTSM), XXVII

Dynamic XPS measurements of ultrathin polyelectrolyte films containing antibacterial Ag–Cu nanoparticles

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

Taner-Camcı, Merve; Suzer, Sefik, E-mail: suzer@fen.bilkent.edu.tr

Ultrathin films consisting of polyelectrolyte layers prepared by layer-by-layer deposition technique and containing also Ag and Cu nanoparticles exhibit superior antibacterial activity toward Escherichia coli. These films have been investigated with XPS measurements under square wave excitation at two different frequencies, in order to further our understanding about the chemical/physical nature of the nanoparticles. Dubbed as dynamical XPS, such measurements bring out similarities and differences among the surface structures by correlating the binding energy shifts of the corresponding XPS peaks. Accordingly, it is observed that the Cu2p, Ag3d of the metal nanoparticles, and S2p of cysteine, the stabilizer and themore » capping agent, exhibit similar shifts. On the other hand, the C1s, N1s, and S2p peaks of the polyelectrolyte layers shift differently. This finding leads us the claim that the Ag and Cu atoms are in a nanoalloy structure, capped with cystein, as opposed to phase separated entities.« less

Transport properties of ultrathin black phosphorus on hexagonal boron nitride

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

Doganov, Rostislav A.; Özyilmaz, Barbaros; Department of Physics, National University of Singapore, 2 Science Drive 3, 117542 Singapore

2015-02-23

Ultrathin black phosphorus, or phosphorene, is a two-dimensional material that allows both high carrier mobility and large on/off ratios. Similar to other atomic crystals, like graphene or layered transition metal dichalcogenides, the transport behavior of few-layer black phosphorus is expected to be affected by the underlying substrate. The properties of black phosphorus have so far been studied on the widely utilized SiO{sub 2} substrate. Here, we characterize few-layer black phosphorus field effect transistors on hexagonal boron nitride—an atomically smooth and charge trap-free substrate. We measure the temperature dependence of the field effect mobility for both holes and electrons and explainmore » the observed behavior in terms of charged impurity limited transport. We find that in-situ vacuum annealing at 400 K removes the p-doping of few-layer black phosphorus on both boron nitride and SiO{sub 2} substrates and reduces the hysteresis at room temperature.« less

Using an ultra-thin non-doped orange emission layer to realize high efficiency white organic light-emitting diodes with low efficiency roll-off

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

Zhu, Liping; Chen, Jiangshan; Ma, Dongge, E-mail: mdg1014@ciac.ac.cn

2014-06-28

By adopting an ultra-thin non-doped orange emission layer sandwiched between two blue emission layers, high efficiency white organic light-emitting diodes (WOLEDs) with reduced efficiency roll-off were fabricated. The optimized devices show a balanced white emission with Internationale de L'Eclairage of (0.41, 0.44) at the luminance of 1000 cd/m{sup 2}, and the maximum power efficiency, current efficiency (CE), and external quantum efficiency reach 63.2 lm/W, 59.3 cd/A, and 23.1%, which slightly shift to 53.4 lm/W, 57.1 cd/A, and 22.2% at 1000 cd/m{sup 2}, respectively, showing low efficiency roll-off. Detailed investigations on the recombination zone and the transient electroluminescence (EL) clearly reveal the EL processes of the ultra-thinmore » non-doped orange emission layer in WOLEDs.« less

Formation and investigation of ultrathin layers of Co2FeSi ferromagnetic alloy synthesized on silicon covered with a CaF2 barrier layer

NASA Astrophysics Data System (ADS)

Grebenyuk, G. S.; Gomoyunova, M. V.; Pronin, I. I.; Vyalikh, D. V.; Molodtsov, S. L.

2016-03-01

Ultrathin (∼2 nm) films of Co2FeSi ferromagnetic alloy were formed on silicon by solid-phase epitaxy and studied in situ. Experiments were carried out in an ultrahigh vacuum (UHV) using substrates of Si(1 1 1) single crystals covered with a 5 nm thick CaF2 barrier layer. The elemental and phase composition as well as the magnetic properties of the synthesized films were analyzed by photoelectron spectroscopy using synchrotron radiation and by magnetic linear dichroism in photoemission of Fe 3p and Co 3p electrons. The study shows that the synthesis of the Co2FeSi ferromagnetic alloy occurs in the temperature range of 200-400 °C. At higher temperatures, the films become island-like and lose their ferromagnetic properties, as the CaF2 barrier layer is unable to prevent a mass transfer between the film and the Si substrate, which violates the stoichiometry of the alloy.

A Comparative Study of AlGaN and InGaN Back-Barriers in Ultrathin-Barrier AlN/GaN Heterostructures

NASA Astrophysics Data System (ADS)

All Abbas, J. M.; Atmaca, G.; Narin, P.; Kutlu, E.; Sarikavak-Lisesivdin, B.; Lisesivdin, S. B.

2017-08-01

Investigations of the effects of back-barrier introduction on the two-dimensional electron gas (2DEG) of ultrathin-barrier AlN/GaN heterostructures with AlGaN and InGaN back-barriers are carried out using self-consistent solutions of 1-dimensional Schrödinger-Poisson equations. Inserted AlGaN and InGaN back-barriers are used to provide a good 2DEG confinement thanks to raising the conduction band edge of GaN buffer with respect to GaN channel layer. Therefore, in this paper the influence of these back-barrier layers on sheet carrier density, 2DEG confinement, and mobility are systematically and comparatively investigated. As a result of calculations, although sheet carrier density is found to decrease with InGaN back-barrier layer, it is not changed with AlGaN back-barrier layer for suggested optimise heterostructures. Obtained results can give some insights for further experimental studies.

An ultra-thin, un-doped NiO hole transporting layer of highly efficient (16.4%) organic-inorganic hybrid perovskite solar cells.

PubMed

Seo, Seongrok; Park, Ik Jae; Kim, Myungjun; Lee, Seonhee; Bae, Changdeuck; Jung, Hyun Suk; Park, Nam-Gyu; Kim, Jin Young; Shin, Hyunjung

2016-06-02

NiO is a wide band gap p-type oxide semiconductor and has potential for applications in solar energy conversion as a hole-transporting layer (HTL). It also has good optical transparency and high chemical stability, and the capability of aligning the band edges to the perovskite (CH3NH3PbI3) layers. Ultra-thin and un-doped NiO films with much less absorption loss were prepared by atomic layer deposition (ALD) with highly precise control over thickness without any pinholes. Thin enough (5-7.5 nm in thickness) NiO films with the thickness of few time the Debye length (LD = 1-2 nm for NiO) show enough conductivities achieved by overlapping space charge regions. The inverted planar perovskite solar cells with NiO films as HTLs exhibited the highest energy conversion efficiency of 16.40% with high open circuit voltage (1.04 V) and fill factor (0.72) with negligible current-voltage hysteresis.

Lateral amorphous selenium metal-insulator-semiconductor-insulator-metal photodetectors using ultrathin dielectric blocking layers for dark current suppression

NASA Astrophysics Data System (ADS)

Chang, Cheng-Yi; Pan, Fu-Ming; Lin, Jian-Siang; Yu, Tung-Yuan; Li, Yi-Ming; Chen, Chieh-Yang

2016-12-01

We fabricated amorphous selenium (a-Se) photodetectors with a lateral metal-insulator-semiconductor-insulator-metal (MISIM) device structure. Thermal aluminum oxide, plasma-enhanced chemical vapor deposited silicon nitride, and thermal atomic layer deposited (ALD) aluminum oxide and hafnium oxide (ALD-HfO2) were used as the electron and hole blocking layers of the MISIM photodetectors for dark current suppression. A reduction in the dark current by three orders of magnitude can be achieved at electric fields between 10 and 30 V/μm. The effective dark current suppression is primarily ascribed to electric field lowering in the dielectric layers as a result of charge trapping in deep levels. Photogenerated carriers in the a-Se layer can be transported across the blocking layers to the Al electrodes via Fowler-Nordheim tunneling because a high electric field develops in the ultrathin dielectric layers under illumination. Since the a-Se MISIM photodetectors have a very low dark current without significant degradation in the photoresponse, the signal contrast is greatly improved. The MISIM photodetector with the ALD-HfO2 blocking layer has an optimal signal contrast more than 500 times the contrast of the photodetector without a blocking layer at 15 V/μm.

“Non-hydrolytic” sol–gel synthesis of molybdenum sulfides

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

Leidich, Saskia; Buechele, Dominique; Lauenstein, Raphael

2016-10-15

Non-hydrolytic sol–gel reactions provide a low temperature solution based synthetic approach to solid-state materials. In this paper, reactions between molybdenum chloride and hexamethyldisilthiane in chloroform were explored, which gave access to both MoS{sub 2} and Mo{sub 2}S{sub 3} after heat treatment of as-recovered amorphous samples to 600–1000 °C. Interesting morphologies were obtained for MoS{sub 2}, ranging from fused spherical particles to well-defined nanoplatelets and nanoflakes. Both 2H- and 3R-MoS{sub 2} were observed, which formed thin hexagonal and triangular platelets, respectively. The platelets exhibited thicknesses of 10–30 nm, which corresponds to 15–50 MoS{sub 2} layers. No attempts to prevent agglomeration weremore » made, however, well separated platelets were observed for many samples. Heating at 1000 °C led to formation of Mo{sub 2}S{sub 3} for samples that showed well-defined MoS{sub 2} at lower temperatures, while less crystalline samples had a tendency to retain the MoS{sub 2} structure. - Graphical abstract: Overlay of variable pressure X-ray diffraction data of Al{sub 2}Mo{sub 3}O{sub 12} collected in a diamond anvil cell. Both subtle and discontinuous phase transitions are clearly observed. - Highlights: • Molybdenum sulfides were prepared by non-hydrolytic sol–gel chemistry. • Nanocrystalline 3R-MoS{sub 2} and 2H-MoS{sub 2}, and microcrystalline Mo{sub 2}S{sub 3} were obtained. • Particle morphology correlated strongly with crystalline phases. • Ultrathin platelets with limited tendency to agglomerate were recovered.« less

In-situ observation of equilibrium transitions in Ni films; agglomeration and impurity effects.

PubMed

Thron, Andrew M; Greene, Peter; Liu, Kai; van Benthem, Klaus

2014-02-01

Dewetting of ultra-thin Ni films deposited on SiO2 layers was observed, in cross-section, by in situ scanning transmission electron microscopy. Holes were observed to nucleate by voids which formed at the Ni/SiO2 interface rather than at triple junctions at the free surface of the Ni film. Ni islands were observed to retract, in attempt to reach equilibrium on the SiO2 layer. SiO2 layers with 120 nm thickness were found to limit in situ heating experiments due to poor thermal conductivity of SiO2. The formation of graphite was observed during the agglomeration of ultra-thin Ni films. Graphite was observed to wet both the free surface and the Ni/SiO2 interface of the Ni islands. Cr forms surface oxide layers on the free surface of the SiO2 layer and the Ni islands. Cr does not prevent the dewetting of Ni, however it will likely alter the equilibrium shape of the Ni islands. © 2013 Published by Elsevier B.V.

Impact of ultra-thin Al2O3-y layers on TiO2-x ReRAM switching characteristics

NASA Astrophysics Data System (ADS)

Trapatseli, Maria; Cortese, Simone; Serb, Alexander; Khiat, Ali; Prodromakis, Themistoklis

2017-05-01

Transition metal-oxide resistive random access memory devices have demonstrated excellent performance in switching speed, versatility of switching and low-power operation. However, this technology still faces challenges like poor cycling endurance, degradation due to high electroforming (EF) switching voltages and low yields. Approaches such as engineering of the active layer by doping or addition of thin oxide buffer layers have been often adopted to tackle these problems. Here, we have followed a strategy that combines the two; we have used ultra-thin Al2O3-y buffer layers incorporated between TiO2-x thin films taking into account both 3+/4+ oxidation states of Al/Ti cations. Our devices were tested by DC and pulsed voltage sweeping and in both cases demonstrated improved switching voltages. We believe that the Al2O3-y layers act as reservoirs of oxygen vacancies which are injected during EF, facilitate a filamentary switching mechanism and provide enhanced filament stability, as shown by the cycling endurance measurements.

Sticker-type Alq(3)-based OLEDs based on printable ultrathin substrates in periodically anchored and suspended configurations.

PubMed

Yun, Su Ok; Hwang, Youngkyu; Park, Jeongpil; Jeong, Yunkyung; Kim, Suk Ho; Noh, Byeong Il; Jung, Hoon Sun; Jang, Hun Soo; Hyun, Yujun; Choa, Sung-Hoon; Ko, Heung Cho

2013-10-18

Introducing two-dimensional post arrays and a water-soluble sacrificial layer between an ultrathin substrate and a handling substrate provides controllability of the interfacial adhesion in a stable manner. The periodically anchored and suspended configuration after the chemical etching process facilitates the development of, for example, printable Alq3 -based OLEDs that can be attached to unconventional surfaces. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ultrathin IBAD MgO films for epitaxial growth on amorphous substrates and sub-50 nm membranes

DOE PAGES

Wang, Siming; Antonakos, C.; Bordel, C.; ...

2016-11-07

Here, a fabrication process has been developed for high energy ion beam assisted deposition (IBAD) biaxial texturing of ultrathin (~1 nm) MgO films, using a high ion-to-atom ratio and post-deposition annealing instead of a homoepitaxial MgO layer. These films serve as the seed layer for epitaxial growth of materials on amorphous substrates such as electron/X-ray transparent membranes or nanocalorimetry devices. Stress measurements and atomic force microscopy of the MgO films reveal decreased stress and surface roughness, while X-ray diffraction of epitaxial overlayers demonstrates the improved crystal quality of films grown epitaxially on IBAD MgO. The process simplifies the synthesis ofmore » IBAD MgO, fundamentally solves the “wrinkle” issue induced by the homoepitaxial layer on sub-50 nm membranes, and enables studies of epitaxial materials in electron/X-ray transmission and nanocalorimetry.« less

Localized Control of Curie Temperature in Perovskite Oxide Film by Capping-Layer-Induced Octahedral Distortion

DOE PAGES

Thomas, S.; Kuiper, B.; Hu, J.; ...

2017-10-27

With reduced dimensionality, it is often easier to modify the properties of ultrathin films than their bulk counterparts. Strain engineering, usually achieved by choosing appropriate substrates, has been proven effective in controlling the properties of perovskite oxide films. An emerging alternative route for developing new multifunctional perovskite is by modification of the oxygen octahedral structure. Here we report the control of structural oxygen octahedral rotation in ultrathin perovskite SrRuO 3 films by the deposition of a SrTiO 3 capping layer, which can be lithographically patterned to achieve local control. Here, using a scanning Sagnac magnetic microscope, we show an increasemore » in the Curie temperature of SrRuO 3 due to the suppression octahedral rotations revealed by the synchrotron x-ray diffraction. Lastly, this capping-layer-based technique may open new possibilities for developing functional oxide materials.« less

Fabrication and stability investigation of ultra-thin transparent and flexible Cu-Ag-Au tri-layer film on PET

NASA Astrophysics Data System (ADS)

Prakasarao, Ch Surya; D'souza, Slavia Deeksha; Hazarika, Pratim; Karthiselva N., S.; Ramesh Babu, R.; Kovendhan, M.; Kumar, R. Arockia; Joseph, D. Paul

2018-04-01

The need for transparent conducting electrodes with high transmittance, low sheet resistance and flexibility to replace Indium Tin Oxide is ever growing. We have deposited and studied the performance of ultra-thin Cu-Ag-Au tri-layer films over a flexible poly-ethylene terephthalate substrate. Scotch tape test showed good adhesion of the metallic film. Transmittance of the tri-layer was around 40 % in visible region. Optical profiler measurements were done to study the surface features. The XRD pattern revealed that film was amorphous. Sheet resistance measured by four probe technique was around 7.7 Ohm/Δ and was stable up to 423 K. The transport parameters by Hall effect showed high conductivity and carrier concentration with a mobility of 5.58 cm2/Vs. Tests performed in an indigenously designed bending unit indicated the films to be stable both mechanically and electrically even after 50,000 bending cycles.

Localized Control of Curie Temperature in Perovskite Oxide Film by Capping-Layer-Induced Octahedral Distortion.

PubMed

Thomas, S; Kuiper, B; Hu, J; Smit, J; Liao, Z; Zhong, Z; Rijnders, G; Vailionis, A; Wu, R; Koster, G; Xia, J

2017-10-27

With reduced dimensionality, it is often easier to modify the properties of ultrathin films than their bulk counterparts. Strain engineering, usually achieved by choosing appropriate substrates, has been proven effective in controlling the properties of perovskite oxide films. An emerging alternative route for developing new multifunctional perovskite is by modification of the oxygen octahedral structure. Here we report the control of structural oxygen octahedral rotation in ultrathin perovskite SrRuO_{3} films by the deposition of a SrTiO_{3} capping layer, which can be lithographically patterned to achieve local control. Using a scanning Sagnac magnetic microscope, we show an increase in the Curie temperature of SrRuO_{3} due to the suppression octahedral rotations revealed by the synchrotron x-ray diffraction. This capping-layer-based technique may open new possibilities for developing functional oxide materials.