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Sample records for direct label-free electrical

  1. Real-time dual-loop electric current measurement for label-free nanofluidic preconcentration chip.

    Science.gov (United States)

    Chung, Pei-Shan; Fan, Yu-Jui; Sheen, Horn-Jiunn; Tian, Wei-Cheng

    2015-01-07

    An electrokinetic trapping (EKT)-based nanofluidic preconcentration device with the capability of label-free monitoring trapped biomolecules through real-time dual-loop electric current measurement was demonstrated. Universal current-voltage (I-V) curves of EKT-based preconcentration devices, consisting of two microchannels connected by ion-selective channels, are presented for functional validation and optimal operation; universal onset current curves indicating the appearance of the EKT mechanism serve as a confirmation of the concentrating action. The EKT mechanism and the dissimilarity in the current curves related to the volume flow rate (Q), diffusion coefficient (D), and diffusion layer (DL) thickness were explained by a control volume model with a five-stage preconcentration process. Different behaviors of the trapped molecular plug were categorized based on four modes associated with different degrees of electroosmotic instability (EOI). A label-free approach to preconcentrating (bio)molecules and monitoring the multibehavior molecular plug was demonstrated through real-time electric current monitoring, rather than through the use of microscope images.

  2. Direct identification of amyloids by label-free quantitative LC-MS

    DEFF Research Database (Denmark)

    Dueholm, Morten Simonsen; Danielsen, Heidi Nolsøe; Hansen, Susan Hove

    Direct identification of amyloids by label-free quantitative LC-MS H. N. Danielsen, S. H. Hansen, F.-A. Herbst, P. H. Nielsen, M. S. Dueholm Amyloids are highly ordered fibrillar protein polymers used by organisms from all domains of life due to their exceptional properties. We have previously...... that have been studied so far have already provided an astonishing demonstration of how the amyloids can be exploited with roles ranging structural components of biofilms, cell envelopes and spore coats to cytotoxins and as reservoirs for quorum-sensing signaling molecules. The identification of novel...... adhesive and therefore bind to pipette tips and other consumables. Pure cultures, large sample volumes and high productivity of amyloids are therefore required for successful purification. We here present a quantitative proteomics technique that allow direct identification of functional amyloid candidates...

  3. Electrochemical direct immobilization of DNA sequences for label-free herpes virus detection

    International Nuclear Information System (INIS)

    Phuong Dinh Tam; Mai Anh Tuan; Tran Trung; Nguyen Duc Chien

    2009-01-01

    DNA sequences/bio-macromolecules of herpes virus (5'-AT CAC CGA CCC GGA GAG GGA C-3') were directly immobilized into polypyrrole matrix by using the cyclic voltammetry method, and grafted onto arrays of interdigitated platinum microelectrodes. The morphology surface of the obtained PPy/DNA of herpes virus composite films was investigated by a FESEM Hitachi-S 4800. Fourier transform infrared spectroscopy (FTIR) was used to characterize the PPy/DNA film and to study the specific interactions that may exist between DNA biomacromolecules and PPy chains. Attempts are made to use these PPy/DNA composite films for label-free herpes virus detection revealed a response time of 60 s in solutions containing as low as 2 nM DNA concentration, and self life of six months when emerged in double distilled water and kept refrigerated.

  4. Electrochemical direct immobilization of DNA sequences for label-free herpes virus detection

    Science.gov (United States)

    Tam, Phuong Dinh; Trung, Tran; Tuan, Mai Anh; Chien, Nguyen Duc

    2009-09-01

    DNA sequences/bio-macromolecules of herpes virus (5'-AT CAC CGA CCC GGA GAG GGA C-3') were directly immobilized into polypyrrole matrix by using the cyclic voltammetry method, and grafted onto arrays of interdigitated platinum microelectrodes. The morphology surface of the obtained PPy/DNA of herpes virus composite films was investigated by a FESEM Hitachi-S 4800. Fourier transform infrared spectroscopy (FTIR) was used to characterize the PPy/DNA film and to study the specific interactions that may exist between DNA biomacromolecules and PPy chains. Attempts are made to use these PPy/DNA composite films for label-free herpes virus detection revealed a response time of 60 s in solutions containing as low as 2 nM DNA concentration, and self life of six months when immerged in double distilled water and kept refrigerated.

  5. Single walled carbon nanotube-based electrical biosensor for the label-free detection of pathogenic bacteria

    DEFF Research Database (Denmark)

    Yoo, S. M.; Baek, Y. K.; Shin, S.

    2016-01-01

    We herein describe the development of a single-walled carbon nanotube (SWNT)-based electrical biosensor consisting of a two-terminal resistor, and report its use for the specific, label-free detection of pathogenic bacteria via changes in conductance. The ability of this biosensor to recognize...

  6. Direct, label-free, selective, and sensitive microbial detection using a bacteriorhodopsin-based photoelectric immunosensor.

    Science.gov (United States)

    Chen, Hsiu-Mei; Jheng, Kai-Ru; Yu, An-Dih

    2017-05-15

    A photoelectric immunosensor using purple membranes (PM) as the transducer, which contains photoactive bacteriorhodopsin, is here first demonstrated for direct and label-free microbial detection. Biotinylated polyclonal antibodies against Escherichia coli were immobilized on a PM-coated electrode through further surface biotinylation and bridging avidin or NeutrAvidin. The photocurrent generated by the antibody-coated sensor was reduced after incubation with E. coli K-12 cultures, with the reduction level increased with the culture populations. The immunosensor prepared via NeutrAvidin exhibited much better selectivity than the one prepared via avidin, recognizing almost none of the tested Gram-positive bacteria. Cultures with populations ranging from 1 to 10 7 CFU/10mL were detected in a single step without any preprocessing. Both AFM and Raman analysis confirmed the layer-by-layer fabrication of the antibody-coated substrates as well as the binding of microorganisms. By investigating the effect of illumination orientation and simulating the photocurrent responses with an equivalent circuit model containing a chemical capacitance, we suggest that the photocurrent reduction was primarily caused by the light-shielding effect of the captured bacteria. Using the current fabrication technique, versatile bacteriorhodopsin-based photoelectric immunosensors can be readily prepared to detect a wide variety of biological cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Label-free direct surface-enhanced Raman scattering (SERS) of nucleic acids (Conference Presentation)

    Science.gov (United States)

    Guerrini, Luca; Morla-Folch, Judit; Gisbert-Quilis, Patricia; Xie, Hainan; Alvarez-Puebla, Ramon

    2016-03-01

    Recently, plasmonic-based biosensing has experienced an unprecedented level of attention, with a particular focus on the nucleic acid detection, offering efficient solutions to engineer simple, fast, highly sensitive sensing platforms while overcoming important limitations of PCR and microarray techniques. In the broad field of plasmonics, surface-enhanced Raman scattering (SERS) spectroscopy has arisen as a powerful analytical tool for detection and structural characterization of biomolecules. Today applications of SERS to nucleic acid analysis largely rely on indirect strategies, which have been demonstrated very effective for pure sensing purposes but completely dismiss the exquisite structural information provided by the direct acquisition of the biomolecular vibrational fingerprint. Contrarily, direct label-free SERS of nucleic acid shows an outstanding potential in terms of chemical-specific information which, however, remained largely unexpressed mainly because of the inherent poor spectral reproducibility and/or limited sensitivity. To address these limitations, we developed a fast and affordable high-throughput screening direct SERS method for gaining detailed genomic information on nucleic acids (DNA and RNA) and for the characterization and quantitative recognition of DNA interactions with exogenous agents. The simple strategy relies on the electrostatic adhesion of DNA/RNA onto positively-charged silver colloids that promotes the nanoparticle aggregation into stable clusters yielding intense and reproducible SERS spectra at picogram level (i.e. the analysis can be performed without the necessity of amplification steps thus providing realistic direct information of the nucleic acid in its native state). We anticipate this method to gain a vast impact and set of applications in different fields, including medical diagnostics, genomic screening, drug discovery, forensic science and even molecular electronics.

  8. Direct, Label-Free, and Rapid Transistor-Based Immunodetection in Whole Serum.

    Science.gov (United States)

    Gutiérrez-Sanz, Óscar; Andoy, Nesha M; Filipiak, Marcin S; Haustein, Natalie; Tarasov, Alexey

    2017-09-22

    Transistor-based biosensors fulfill many requirements posed upon transducers for future point-of-care diagnostic devices such as scalable fabrication and label-free and real-time quantification of chemical and biological species with high sensitivity. However, the short Debye screening length in physiological samples (transistor-based biosensors, without the need for sample pretreatment, labeling, or washing steps. The presented sensor is low-cost, can be easily integrated into portable diagnostics devices, and offers a competitive performance compared to state-of-the-art central laboratory analyzers.

  9. Real-Time Label-Free Direct Electronic Monitoring of Topoisomerase Enzyme Binding Kinetics on Graphene.

    Science.gov (United States)

    Zuccaro, Laura; Tesauro, Cinzia; Kurkina, Tetiana; Fiorani, Paola; Yu, Hak Ki; Knudsen, Birgitta R; Kern, Klaus; Desideri, Alessandro; Balasubramanian, Kannan

    2015-11-24

    Monolayer graphene field-effect sensors operating in liquid have been widely deployed for detecting a range of analyte species often under equilibrium conditions. Here we report on the real-time detection of the binding kinetics of the essential human enzyme, topoisomerase I interacting with substrate molecules (DNA probes) that are immobilized electrochemically on to monolayer graphene strips. By monitoring the field-effect characteristics of the graphene biosensor in real-time during the enzyme-substrate interactions, we are able to decipher the surface binding constant for the cleavage reaction step of topoisomerase I activity in a label-free manner. Moreover, an appropriate design of the capture probes allows us to distinctly follow the cleavage step of topoisomerase I functioning in real-time down to picomolar concentrations. The presented results are promising for future rapid screening of drugs that are being evaluated for regulating enzyme activity.

  10. A label-free impedimetric immunosensor for direct determination of the textile dye Disperse Orange 1.

    Science.gov (United States)

    Yang, Jing; da Rocha, Carolina Gomes; Wang, Shengfu; Ferreira, Antonio Aparecido Pupim; Yamanaka, Hideko

    2015-09-01

    A strategy for a label-free impedimetric immunosensor is described for detection of the textile dye Disperse Orange 1 (DO1). The compounds 1,12-diaminododecane (DADD) and then 1,7-diaminoheptane (DAH) were firstly successively grafted onto a glassy carbon electrode (GCE) surface by electro-oxidation of one amino group, while the other terminal amino group was modified with the antibody anti-DO1. The construction process of the immunosensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy and capacitance measurements. The electron transfer resistance (Rct) exhibited an effective response to the affinity between the immobilized antibody and the antigen in solution. The linear range for the target compound was from 5.0 nmol L(-1) to 0.5 μmol L(-1) (R=0.9980), and the limit of detection (LOD) was 7.56 nmol L(-1). The proposed impedimetric immunosensor has the advantages of simplicity, cost-effectiveness, and sensitivity. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Label-free electrical determination of trypsin activity by a silicon-on-insulator based thin film resistor.

    Science.gov (United States)

    Neff, Petra A; Serr, Andreas; Wunderlich, Bernhard K; Bausch, Andreas R

    2007-10-08

    A silicon-on-insulator (SOI) based thin film resistor is employed for the label-free determination of enzymatic activity. We demonstrate that enzymes, which cleave biological polyelectrolyte substrates, can be detected by the sensor. As an application, we consider the serine endopeptidase trypsin, which cleaves poly-L-lysine (PLL). We show that PLL adsorbs quasi-irreversibly to the sensor and is digested by trypsin directly at the sensor surface. The created PLL fragments are released into the bulk solution due to kinetic reasons. This results in a measurable change of the surface potential allowing for the determination of trypsin concentrations down to 50 ng mL(-1). Chymotrypsin is a similar endopeptidase with a different specificity, which cleaves PLL with a lower efficiency as compared to trypsin. The activity of trypsin is analyzed quantitatively employing a kinetic model for enzyme-catalyzed surface reactions. Moreover, we have demonstrated the specific inactivation of trypsin by a serine protease inhibitor, which covalently binds to the active site of the enzyme.

  12. Ligand-Directed Functional Selectivity at the Mu Opioid Receptor Revealed by Label-Free Integrative Pharmacology On-Target

    Science.gov (United States)

    Morse, Megan; Tran, Elizabeth; Sun, Haiyan; Levenson, Robert; Fang, Ye

    2011-01-01

    Development of new opioid drugs that provide analgesia without producing dependence is important for pain treatment. Opioid agonist drugs exert their analgesia effects primarily by acting at the mu opioid receptor (MOR) sites. High-resolution differentiation of opioid ligands is crucial for the development of new lead drug candidates with better tolerance profiles. Here, we use a label-free integrative pharmacology on-target (iPOT) approach to characterize the functional selectivity of a library of known opioid ligands for the MOR. This approach is based on the ability to detect dynamic mass redistribution (DMR) arising from the activation of the MOR in living cells. DMR assays were performed in HEK-MOR cells with and without preconditioning with probe molecules using label-free resonant waveguide grating biosensors, wherein the probe molecules were used to modify the activity of specific signaling proteins downstream the MOR. DMR signals obtained were then translated into high resolution heat maps using similarity analysis based on a numerical matrix of DMR parameters. Our data indicate that the iPOT approach clearly differentiates functional selectivity for distinct MOR signaling pathways among different opioid ligands, thus opening new avenues to discover and quantify the functional selectivity of currently used and novel opioid receptor drugs. PMID:22003401

  13. Graphene-interfaced electrical biosensor for label-free and sensitive detection of foodborne pathogenic E. coli O157:H7.

    Science.gov (United States)

    Pandey, Ashish; Gurbuz, Yasar; Ozguz, Volkan; Niazi, Javed H; Qureshi, Anjum

    2017-05-15

    E. coli O157:H7 is an enterohemorrhagic bacteria responsible for serious foodborne outbreaks that causes diarrhoea, fever and vomiting in humans. Recent foodborne E. coli outbreaks has left a serious concern to public health. Therefore, there is an increasing demand for a simple, rapid and sensitive method for pathogen detection in contaminated foods. In this study, we developed a label-free electrical biosensor interfaced with graphene for sensitive detection of pathogenic bacteria. This biosensor was fabricated by interfacing graphene with interdigitated microelectrodes of capacitors that were biofunctionalized with E. coli O157:H7 specific antibodies for sensitive pathogenic bacteria detection. Here, graphene nanostructures on the sensor surface provided superior chemical properties such as high carrier mobility and biocompatibility with antibodies and bacteria. The sensors transduced the signal based on changes in dielectric properties (capacitance) through (i) polarization of captured cell-surface charges, (ii) cells' internal bioactivity, (iii) cell-wall's electronegativity or dipole moment and their relaxation and (iv) charge carrier mobility of graphene that modulated the electrical properties once the pathogenic E. coli O157:H7 captured on the sensor surface. Sensitive capacitance changes thus observed with graphene based capacitors were specific to E. coli O157:H7 strain with a sensitivity as low as 10-100 cells/ml. The proposed graphene based electrical biosensor provided advantages of speed, sensitivity, specificity and in-situ bacterial detection with no chemical mediators, represents a versatile approach for detection of a wide variety of other pathogens. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Microneedle pH Sensor: Direct, Label-Free, Real-Time Detection of Cerebrospinal Fluid and Bladder pH.

    Science.gov (United States)

    Mani, Ganesh Kumar; Miyakoda, Kousei; Saito, Asuka; Yasoda, Yutaka; Kajiwara, Kagemasa; Kimura, Minoru; Tsuchiya, Kazuyoshi

    2017-07-05

    Acid-base homeostasis (body pH) inside the body is precisely controlled by the kidneys and lungs and buffer systems, such that even a minor pH change could severely affect many organs. Blood and urine pH tests are common in day-to-day clinical trials and require little effort for diagnosis. There is always a great demand for in vivo testing to understand more about body metabolism and to provide effective diagnosis and therapy. In this article, we report the simple fabrication of microneedle-based direct, label-free, and real-time pH sensors. The reference and working electrodes were Ag/AgCl thick films and ZnO thin films on tungsten (W) microneedles, respectively. The morphological and structural characteristics of microneedles were carefully investigated through various analytical methods. The developed sensor exhibited a Nernstian response of -46 mV/pH. Different conditions were used to test the sensor to confirm their accuracy and stability, such as various buffer solutions, with respect to time, and we compared the reading with commercial pH electrodes. Besides that, the fabricated microneedle sensor ability is proven by in vivo testing in mouse cerebrospinal fluid (CSF) and bladders. The pH sensor procedure reported here is totally reversible, and results were reproducible after several rounds of testing.

  15. Lab-on-a-chip for label free biological semiconductor analysis of Staphylococcal Enterotoxin B

    NARCIS (Netherlands)

    Yang, Minghui; Sun, Steven; Bruck, Hugh Alan; Kostov, Yordan; Rasooly, Avraham

    2010-01-01

    We describe a new lab-on-a-chip (LOC) which utilizes a biological semiconductor (BSC) transducer for label free analysis of Staphylococcal Enterotoxin B (SEB) (or other biological interactions) directly and electronically. BSCs are new transducers based on electrical percolation through a

  16. Label-free drug discovery

    Directory of Open Access Journals (Sweden)

    Ye eFang

    2014-03-01

    Full Text Available Current drug discovery is dominated by label-dependent molecular approaches, which screen drugs in the context of a predefined and target-based hypothesis in vitro. Given that target-based discovery has not transformed the industry, phenotypic screen that identifies drugs based on a specific phenotype of cells, tissues, or animals has gained renewed interest. However, owing to the intrinsic complexity in drug-target interactions, there is often a significant gap between the phenotype screened and the ultimate molecular mechanism of action sought. This paper presents a label-free strategy for early drug discovery. This strategy combines label-free cell phenotypic profiling with computational approaches, and holds promise to bridge the gap by offering a kinetic and holistic representation of the functional consequences of drugs in disease relevant cells that is amenable to mechanistic deconvolution.

  17. Multimodal label-free microscopy

    Directory of Open Access Journals (Sweden)

    Nicolas Pavillon

    2014-09-01

    Full Text Available This paper reviews the different multimodal applications based on a large extent of label-free imaging modalities, ranging from linear to nonlinear optics, while also including spectroscopic measurements. We put specific emphasis on multimodal measurements going across the usual boundaries between imaging modalities, whereas most multimodal platforms combine techniques based on similar light interactions or similar hardware implementations. In this review, we limit the scope to focus on applications for biology such as live cells or tissues, since by their nature of being alive or fragile, we are often not free to take liberties with the image acquisition times and are forced to gather the maximum amount of information possible at one time. For such samples, imaging by a given label-free method usually presents a challenge in obtaining sufficient optical signal or is limited in terms of the types of observable targets. Multimodal imaging is then particularly attractive for these samples in order to maximize the amount of measured information. While multimodal imaging is always useful in the sense of acquiring additional information from additional modes, at times it is possible to attain information that could not be discovered using any single mode alone, which is the essence of the progress that is possible using a multimodal approach.

  18. Label-free photoacoustic nanoscopy

    Science.gov (United States)

    Danielli, Amos; Maslov, Konstantin; Garcia-Uribe, Alejandro; Winkler, Amy M.; Li, Chiye; Wang, Lidai; Chen, Yun; Dorn, Gerald W.; Wang, Lihong V.

    2014-08-01

    Super-resolution microscopy techniques-capable of overcoming the diffraction limit of light-have opened new opportunities to explore subcellular structures and dynamics not resolvable in conventional far-field microscopy. However, relying on staining with exogenous fluorescent markers, these techniques can sometimes introduce undesired artifacts to the image, mainly due to large tagging agent sizes and insufficient or variable labeling densities. By contrast, the use of endogenous pigments allows imaging of the intrinsic structures of biological samples with unaltered molecular constituents. Here, we report label-free photoacoustic (PA) nanoscopy, which is exquisitely sensitive to optical absorption, with an 88 nm resolution. At each scanning position, multiple PA signals are successively excited with increasing laser pulse energy. Because of optical saturation or nonlinear thermal expansion, the PA amplitude depends on the nonlinear incident optical fluence. The high-order dependence, quantified by polynomial fitting, provides super-resolution imaging with optical sectioning. PA nanoscopy is capable of super-resolution imaging of either fluorescent or nonfluorescent molecules.

  19. Hybrid Integrated Label-Free Chemical and Biological Sensors

    OpenAIRE

    Mehrabani, Simin; Maker, Ashley J.; Armani, Andrea M.

    2014-01-01

    Label-free sensors based on electrical, mechanical and optical transduction methods have potential applications in numerous areas of society, ranging from healthcare to environmental monitoring. Initial research in the field focused on the development and optimization of various sensor platforms fabricated from a single material system, such as fiber-based optical sensors and silicon nanowire-based electrical sensors. However, more recent research efforts have explored designing sensors fabri...

  20. Label-free electrical sensing of bacteria in eye wash samples: A step towards point-of-care detection of pathogens in patients with infectious keratitis.

    Science.gov (United States)

    Pandya, Hardik J; Kanakasabapathy, Manoj Kumar; Verma, Saloni; Chug, Manjyot Kaur; Memic, Adnan; Gadjeva, Mihaela; Shafiee, Hadi

    2017-05-15

    The diagnosis of keratitis is based on visual exam, tissue cytology, and standard microbial culturing to determine the type of the infectious pathogen. To prescribe appropriate therapy, it is important to distinguish between bacterial, fungal, and viral keratitis, as the treatments are quite different. Diagnosis of the causative organism has a substantial prognostic importance. Further, timely knowledge of the nature of the pathogen is also critical to adapt therapy in patients unresponsive to empiric treatment options, which occurs in 10% of all cases. Currently, the identification of the nature of the pathogen that causes keratitis is achieved via microbial culture screening, which is laboratory-based, expensive, and time-consuming. The most frequent pathogens that cause the corneal ulcers are P. aeruginosa and S. aureus. Here, we report a microchip for rapid (<1h) detection of P. aeruginosa (6294), S. aureus(LAC), through on-chip electrical sensing of bacterial lysate. We evaluated the microchip with spiked samples of PBS with bacteria concentration between 10 1 to 10 8 CFU/mL. The least diluted bacteria concentration in bacteria-spiked samples with statistically significant impedance change was 10 CFU/mL. We further validated our assay by comparing our microchip results with the standard culture-based methods using eye washes obtained from 13 infected mice. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Emerging applications of label-free optical biosensors

    Science.gov (United States)

    Zanchetta, Giuliano; Lanfranco, Roberta; Giavazzi, Fabio; Bellini, Tommaso; Buscaglia, Marco

    2017-01-01

    Innovative technical solutions to realize optical biosensors with improved performance are continuously proposed. Progress in material fabrication enables developing novel substrates with enhanced optical responses. At the same time, the increased spectrum of available biomolecular tools, ranging from highly specific receptors to engineered bioconjugated polymers, facilitates the preparation of sensing surfaces with controlled functionality. What remains often unclear is to which extent this continuous innovation provides effective breakthroughs for specific applications. In this review, we address this challenging question for the class of label-free optical biosensors, which can provide a direct signal upon molecular binding without using secondary probes. Label-free biosensors have become a consolidated approach for the characterization and screening of molecular interactions in research laboratories. However, in the last decade, several examples of other applications with high potential impact have been proposed. We review the recent advances in label-free optical biosensing technology by focusing on the potential competitive advantage provided in selected emerging applications, grouped on the basis of the target type. In particular, direct and real-time detection allows the development of simpler, compact, and rapid analytical methods for different kinds of targets, from proteins to DNA and viruses. The lack of secondary interactions facilitates the binding of small-molecule targets and minimizes the perturbation in single-molecule detection. Moreover, the intrinsic versatility of label-free sensing makes it an ideal platform to be integrated with biomolecular machinery with innovative functionality, as in case of the molecular tools provided by DNA nanotechnology.

  2. Functionalized nanopipettes: toward label-free, single cell biosensors

    OpenAIRE

    Actis, Paolo; Mak, Andy C.; Pourmand, Nader

    2010-01-01

    Nanopipette technology has been proven to be a label-free biosensor capable of identifying DNA and proteins. The nanopipette can include specific recognition elements for analyte discrimination based on size, shape, and charge density. The fully electrical read-out and the ease and low-cost fabrication are unique features that give this technology an enormous potential. Unlike other biosensing platforms, nanopipettes can be precisely manipulated with submicron accuracy and used to study singl...

  3. Performance limitations of label-free sensors in molecular diagnosis using complex samples

    Science.gov (United States)

    Varma, Manoj

    2016-03-01

    Label-free biosensors promised a paradigm involving direct detection of biomarkers from complex samples such as serum without requiring multistep sample processing typical of labelled methods such as ELISA or immunofluorescence assays. Label-free sensors have witnessed decades of development with a veritable zoo of techniques available today exploiting a multitude of physical effects. It is appropriate now to critically assess whether label-free technologies have succeeded in delivering their promise with respect to diagnostic applications, particularly, ambitious goals such as early cancer detection using serum biomarkers, which require low limits of detection (LoD). Comparison of nearly 120 limits of detection (LoD) values reported by labelled and label-free sensing approaches over a wide range of detection techniques and target molecules in serum revealed that labeled techniques achieve 2-3 orders of magnitude better LoDs. Data from experiments where labelled and label-free assays were performed simultaneously using the same assay parameters also confirm that the LoD achieved by labelled techniques is 2 to 3 orders of magnitude better than that by label-free techniques. Furthermore, label-free techniques required significant signal amplification, for e.g. using nanoparticle conjugated secondary antibodies, to achieve LoDs comparable to labelled methods substantially deviating from the original "direct detection" paradigm. This finding has important implications on the practical limits of applying label-free detection methods for molecular diagnosis.

  4. Label-Free Biosensors for Cell Biology

    Directory of Open Access Journals (Sweden)

    Ye Fang

    2011-01-01

    Full Text Available Label-free biosensors for studying cell biology have finally come of age. Recent developments have advanced the biosensors from low throughput and high maintenance research tools to high throughput and low maintenance screening platforms. In parallel, the biosensors have evolved from an analytical tool solely for molecular interaction analysis to powerful platforms for studying cell biology at the whole cell level. This paper presents historical development, detection principles, and applications in cell biology of label-free biosensors. Future perspectives are also discussed.

  5. Hybrid Integrated Label-Free Chemical and Biological Sensors

    Directory of Open Access Journals (Sweden)

    Simin Mehrabani

    2014-03-01

    Full Text Available Label-free sensors based on electrical, mechanical and optical transduction methods have potential applications in numerous areas of society, ranging from healthcare to environmental monitoring. Initial research in the field focused on the development and optimization of various sensor platforms fabricated from a single material system, such as fiber-based optical sensors and silicon nanowire-based electrical sensors. However, more recent research efforts have explored designing sensors fabricated from multiple materials. For example, synthetic materials and/or biomaterials can also be added to the sensor to improve its response toward analytes of interest. By leveraging the properties of the different material systems, these hybrid sensing devices can have significantly improved performance over their single-material counterparts (better sensitivity, specificity, signal to noise, and/or detection limits. This review will briefly discuss some of the methods for creating these multi-material sensor platforms and the advances enabled by this design approach.

  6. Hybrid integrated label-free chemical and biological sensors.

    Science.gov (United States)

    Mehrabani, Simin; Maker, Ashley J; Armani, Andrea M

    2014-03-26

    Label-free sensors based on electrical, mechanical and optical transduction methods have potential applications in numerous areas of society, ranging from healthcare to environmental monitoring. Initial research in the field focused on the development and optimization of various sensor platforms fabricated from a single material system, such as fiber-based optical sensors and silicon nanowire-based electrical sensors. However, more recent research efforts have explored designing sensors fabricated from multiple materials. For example, synthetic materials and/or biomaterials can also be added to the sensor to improve its response toward analytes of interest. By leveraging the properties of the different material systems, these hybrid sensing devices can have significantly improved performance over their single-material counterparts (better sensitivity, specificity, signal to noise, and/or detection limits). This review will briefly discuss some of the methods for creating these multi-material sensor platforms and the advances enabled by this design approach.

  7. Hybrid Integrated Label-Free Chemical and Biological Sensors

    Science.gov (United States)

    Mehrabani, Simin; Maker, Ashley J.; Armani, Andrea M.

    2014-01-01

    Label-free sensors based on electrical, mechanical and optical transduction methods have potential applications in numerous areas of society, ranging from healthcare to environmental monitoring. Initial research in the field focused on the development and optimization of various sensor platforms fabricated from a single material system, such as fiber-based optical sensors and silicon nanowire-based electrical sensors. However, more recent research efforts have explored designing sensors fabricated from multiple materials. For example, synthetic materials and/or biomaterials can also be added to the sensor to improve its response toward analytes of interest. By leveraging the properties of the different material systems, these hybrid sensing devices can have significantly improved performance over their single-material counterparts (better sensitivity, specificity, signal to noise, and/or detection limits). This review will briefly discuss some of the methods for creating these multi-material sensor platforms and the advances enabled by this design approach. PMID:24675757

  8. Label-free protein detection using a microfluidic Coulter-counter device

    DEFF Research Database (Denmark)

    Rodriguez-Trujíllo, Romén; Ajine, Mohammad Akram; Orzan, A.

    2014-01-01

    A new method for measuring specific protein concentrations in solutions has been developed. The technique makes use of the Coulter effect for detecting and sizing of micro-scaled objects suspended in a buffer fluid. The method is completely label-free as it is only based on the electrical readout...

  9. Label-free isolation of circulating tumor cells in microfluidic devices: Current research and perspectives.

    Science.gov (United States)

    Cima, Igor; Wen Yee, Chay; Iliescu, Florina S; Phyo, Wai Min; Lim, Kiat Hon; Iliescu, Ciprian; Tan, Min Han

    2013-01-01

    This review will cover the recent advances in label-free approaches to isolate and manipulate circulating tumor cells (CTCs). In essence, label-free approaches do not rely on antibodies or biological markers for labeling the cells of interest, but enrich them using the differential physical properties intrinsic to cancer and blood cells. We will discuss technologies that isolate cells based on their biomechanical and electrical properties. Label-free approaches to analyze CTCs have been recently invoked as a valid alternative to "marker-based" techniques, because classical epithelial and tumor markers are lost on some CTC populations and there is no comprehensive phenotypic definition for CTCs. We will highlight the advantages and drawbacks of these technologies and the status on their implementation in the clinics.

  10. Label-free identification of activated T lymphocytes through tridimensional microsensors on chip.

    Science.gov (United States)

    Rollo, Enrica; Tenaglia, Enrico; Genolet, Raphaël; Bianchi, Elena; Harari, Alexandre; Coukos, George; Guiducci, Carlotta

    2017-08-15

    Label-free approaches to assess cell properties ideally suit the requirements of cell-based therapeutics, since they permit to characterize cells with minimal perturbation and manipulation, at the benefit of sample recovery and re-employment for treatment. For this reason, label-free techniques would find sensible application in adoptive T cell-based immunotherapy. In this work, we describe the label-free and single-cell detection of in vitro activated T lymphocytes in flow through an electrical impedance-based setup. We describe a novel platform featuring 3D free-standing microelectrodes presenting passive upstream and downstream extensions and integrated into microfluidic channels. We employ such device to measure the impedance change associated with T cell activation at electrical frequencies maximizing the difference between non-activated and activated T cells. Finally, we harness the impedance signature of unstimulated T cells to set a boundary separating activated and non-activated clones, so to characterize the selectivity and specificity of the system. In conclusion, the strategy here proposed highlights the possible employment of impedance to assess T cell activation in label-free. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Label-free SERS in biological and biomedical applications: Recent progress, current challenges and opportunities.

    Science.gov (United States)

    Zheng, Xiao-Shan; Jahn, Izabella Jolan; Weber, Karina; Cialla-May, Dana; Popp, Jürgen

    2018-05-15

    To achieve an insightful look within biomolecular processes on the cellular level, the development of diseases as well as the reliable detection of metabolites and pathogens, a modern analytical tool is needed that is highly sensitive, molecular-specific and exhibits fast detection. Surface-enhanced Raman spectroscopy (SERS) is known to meet these requirements and, within this review article, the recent progress of label-free SERS in biological and biomedical applications is summarized and discussed. This includes the detection of biomolecules such as metabolites, nucleic acids and proteins. Further, the characterization and identification of microorganisms has been achieved by label-free SERS-based approaches. Eukaryotic cells can be characterized by SERS in order to gain information about the outer cell wall or to detect intracellular molecules and metabolites. The potential of SERS for medically relevant detection schemes is emphasized by the label-free detection of tissue, the investigation of body fluids as well as applications for therapeutic and illicit drug monitoring. The review article is concluded with an evaluation of the recent progress and current challenges in order to highlight the direction of label-free SERS in the future. Copyright © 2018. Published by Elsevier B.V.

  12. Label-free SERS in biological and biomedical applications: Recent progress, current challenges and opportunities

    Science.gov (United States)

    Zheng, Xiao-Shan; Jahn, Izabella Jolan; Weber, Karina; Cialla-May, Dana; Popp, Jürgen

    2018-05-01

    To achieve an insightful look within biomolecular processes on the cellular level, the development of diseases as well as the reliable detection of metabolites and pathogens, a modern analytical tool is needed that is highly sensitive, molecular-specific and exhibits fast detection. Surface-enhanced Raman spectroscopy (SERS) is known to meet these requirements and, within this review article, the recent progress of label-free SERS in biological and biomedical applications is summarized and discussed. This includes the detection of biomolecules such as metabolites, nucleic acids and proteins. Further, the characterization and identification of microorganisms has been achieved by label-free SERS-based approaches. Eukaryotic cells can be characterized by SERS in order to gain information about the outer cell wall or to detect intracellular molecules and metabolites. The potential of SERS for medically relevant detection schemes is emphasized by the label-free detection of tissue, the investigation of body fluids as well as applications for therapeutic and illicit drug monitoring. The review article is concluded with an evaluation of the recent progress and current challenges in order to highlight the direction of label-free SERS in the future.

  13. Label-free all-electronic biosensing in microfluidic systems

    Science.gov (United States)

    Stanton, Michael A.

    Label-free, all-electronic detection techniques offer great promise for advancements in medical and biological analysis. Electrical sensing can be used to measure both interfacial and bulk impedance changes in conducting solutions. Electronic sensors produced using standard microfabrication processes are easily integrated into microfluidic systems. Combined with the sensitivity of radiofrequency electrical measurements, this approach offers significant advantages over competing biological sensing methods. Scalable fabrication methods also provide a means of bypassing the prohibitive costs and infrastructure associated with current technologies. We describe the design, development and use of a radiofrequency reflectometer integrated into a microfluidic system towards the specific detection of biologically relevant materials. We developed a detection protocol based on impedimetric changes caused by the binding of antibody/antigen pairs to the sensing region. Here we report the surface chemistry that forms the necessary capture mechanism. Gold-thiol binding was utilized to create an ordered alkane monolayer on the sensor surface. Exposed functional groups target the N-terminus, affixing a protein to the monolayer. The general applicability of this method lends itself to a wide variety of proteins. To demonstrate specificity, commercially available mouse anti- Streptococcus Pneumoniae monoclonal antibody was used to target the full-length recombinant pneumococcal surface protein A, type 2 strain D39 expressed by Streptococcus Pneumoniae. We demonstrate the RF response of the sensor to both the presence of the surface decoration and bound SPn cells in a 1x phosphate buffered saline solution. The combined microfluidic sensor represents a powerful platform for the analysis and detection of cells and biomolecules.

  14. Label-free virus detection using silicon photonic microring resonators.

    Science.gov (United States)

    McClellan, Melinda S; Domier, Leslie L; Bailey, Ryan C

    2012-01-15

    Viruses represent a continual threat to humans through a number of mechanisms, which include disease, bioterrorism, and destruction of both plant and animal food resources. Many contemporary techniques used for the detection of viruses and viral infections suffer from limitations such as the need for extensive sample preparation or the lengthy window between infection and measurable immune response, for serological methods. In order to develop a method that is fast, cost-effective, and features reduced sample preparation compared to many other virus detection methods, we report the application of silicon photonic microring resonators for the direct, label-free detection of intact viruses in both purified samples as well as in a complex, real-world analytical matrix. As a model system, we demonstrate the quantitative detection of Bean pod mottle virus, a pathogen of great agricultural importance, with a limit of detection of 10 ng/mL. By simply grinding a small amount of leaf sample in buffer with a mortar and pestle, infected leaves can be identified over a healthy control with a total analysis time of less than 45 min. Given the inherent scalability and multiplexing capability of the semiconductor-based technology, we feel that silicon photonic microring resonators are well-positioned as a promising analytical tool for a number of viral detection applications. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Functionalizable low-fouling coatings for label-free biosensing in complex biological media: advances and applications

    Czech Academy of Sciences Publication Activity Database

    Vaisocherová, Hana; Brynda, Eduard; Homola, Jiří

    2015-01-01

    Roč. 407, č. 14 (2015), s. 3927-3953 ISSN 1618-2642 R&D Projects: GA ČR(CZ) GBP205/12/G118 Institutional support: RVO:67985882 ; RVO:61389013 Keywords : Label-free optical biosensors * Polymer brushes * Blood plasma Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 3.125, year: 2015

  16. Label-free detection of C-reactive protein using an electrochemical DNA immunoassay

    OpenAIRE

    Songjaroen, Temsiri; Feeny, Rachel M.; Mensack, Meghan M.; Laiwattanapaisal, Wanida; Henry, Charles S.

    2016-01-01

    A label-free electrochemical immunoassay that combines DNA-directed immobilization (DDI) with electrochemical impedance spectroscopy (EIS) on microwire sensors is reported for the detection of C-reactive protein (CRP). CRP is an acute-phase protein that is strongly correlated with systemic inflammation. Since inflammation plays a role in pathogenesis of cardiovascular diseases, CRP can be used to predict the likelihood of coronary events. To demonstrate the new chemistry, 25-μm Au electrodes ...

  17. Label-free monitoring of diffusion in microfluidics

    DEFF Research Database (Denmark)

    Sørensen, Kristian Tølbøl; Kristensen, Anders

    2017-01-01

    Label-free, real-time detection of concentration gradients is demonstrated in a microfluidic H-filter, using an integrated photonic crystal slab sensor to monitor sample refractive index with spatial resolution. The recorded diffusion profiles reveal root-mean-square diffusion lengths for non...

  18. Deep Learning in Label-free Cell Classification

    Science.gov (United States)

    Chen, Claire Lifan; Mahjoubfar, Ata; Tai, Li-Chia; Blaby, Ian K.; Huang, Allen; Niazi, Kayvan Reza; Jalali, Bahram

    2016-03-01

    Label-free cell analysis is essential to personalized genomics, cancer diagnostics, and drug development as it avoids adverse effects of staining reagents on cellular viability and cell signaling. However, currently available label-free cell assays mostly rely only on a single feature and lack sufficient differentiation. Also, the sample size analyzed by these assays is limited due to their low throughput. Here, we integrate feature extraction and deep learning with high-throughput quantitative imaging enabled by photonic time stretch, achieving record high accuracy in label-free cell classification. Our system captures quantitative optical phase and intensity images and extracts multiple biophysical features of individual cells. These biophysical measurements form a hyperdimensional feature space in which supervised learning is performed for cell classification. We compare various learning algorithms including artificial neural network, support vector machine, logistic regression, and a novel deep learning pipeline, which adopts global optimization of receiver operating characteristics. As a validation of the enhanced sensitivity and specificity of our system, we show classification of white blood T-cells against colon cancer cells, as well as lipid accumulating algal strains for biofuel production. This system opens up a new path to data-driven phenotypic diagnosis and better understanding of the heterogeneous gene expressions in cells.

  19. Mutagenic Potential of Direct Current Electric Fields

    National Research Council Canada - National Science Library

    Obringer, John

    1997-01-01

    .... EMF can be subdivided into either electric fields (E-field) or magnetic fields (B-field). Our research used a reverse mutagenesis bacteriophage T4D model to quantitatively study the effects of direct current E-fields...

  20. Label-free optical detection of biomolecular translocation through nanopore arrays.

    Science.gov (United States)

    Ivankin, Andrey; Henley, Robert Y; Larkin, Joseph; Carson, Spencer; Toscano, Michael L; Wanunu, Meni

    2014-10-28

    In recent years, nanopores have emerged as exceptionally promising single-molecule sensors due to their ability to detect biomolecules at subfemtomole levels in a label-free manner. Development of a high-throughput nanopore-based biosensor requires multiplexing of nanopore measurements. Electrical detection, however, poses a challenge, as each nanopore circuit must be electrically independent, which requires complex nanofluidics and embedded electrodes. Here, we present an optical method for simultaneous measurements of the ionic current across an array of solid-state nanopores, requiring no additional fabrication steps. Proof-of-principle experiments are conducted that show simultaneous optical detection and characterization of ssDNA and dsDNA using an array of pores. Through a comparison with electrical measurements, we show that optical measurements are capable of accessing equivalent transmembrane current information.

  1. Responsive Hydrogels for Label-Free Signal Transduction within Biosensors

    Directory of Open Access Journals (Sweden)

    Kamila Gawel

    2010-04-01

    Full Text Available Hydrogels have found wide application in biosensors due to their versatile nature. This family of materials is applied in biosensing either to increase the loading capacity compared to two-dimensional surfaces, or to support biospecific hydrogel swelling occurring subsequent to specific recognition of an analyte. This review focuses on various principles underpinning the design of biospecific hydrogels acting through various molecular mechanisms in transducing the recognition event of label-free analytes. Towards this end, we describe several promising hydrogel systems that when combined with the appropriate readout platform and quantitative approach could lead to future real-life applications.

  2. Label-Free Raman Imaging to Monitor Breast Tumor Signatures

    Science.gov (United States)

    Ciubuc, John

    Methods built on Raman spectroscopy have shown major potential in describing and discriminating between malignant and benign specimens. Accurate, real-time medical diagnosis benefits in substantial improvements through this vibrational optical method. Not only is acquisition of data possible in milliseconds and analysis in minutes, Raman allows concurrent detection and monitoring of all biological components. Besides validating a significant Raman signature distinction between non-tumorigenic (MCF-10A) and tumorigenic (MCF-7) breast epithelial cells, this study reveals a label-free method to assess overexpression of epidermal growth factor receptors (EGFR) in tumor cells. EGFR overexpression sires Raman features associated with phosphorylated threonine and serine, and modifications of DNA/RNA characteristics. Investigations by gel electrophoresis reveal EGF induction of phosphorylated Akt, agreeing with the Raman results. The analysis presented is a vital step toward Raman-based evaluation of EGF receptors in breast cancer cells. With the goal of clinically applying Raman-guided methods for diagnosis of breast tumors, the current results lay the basis for proving label-free optical alternatives in making prognosis of the disease.

  3. Direct Determination of a Small-Molecule Drug, Valproic Acid, by an Electrically-Detected Microcantilever Biosensor for Personalized Diagnostics

    Directory of Open Access Journals (Sweden)

    Long-Sun Huang

    2015-01-01

    Full Text Available Direct, small-molecule determination of the antiepileptic drug, valproic acid, was investigated by a label-free, nanomechanical biosensor. Valproic acid has long been used as an antiepileptic medication, which is administered through therapeutic drug monitoring and has a narrow therapeutic dosage range of 50–100 μg·mL−1 in blood or serum. Unlike labeled and clinically-used measurement techniques, the label-free, electrical detection microcantilever biosensor can be miniaturized and simplified for use in portable or hand-held point-of-care platforms or personal diagnostic tools. A micromachined microcantilever sensor was packaged into the micro-channel of a fluidic system. The measurement of the antiepileptic drug, valproic acid, in phosphate-buffered saline and serum used a single free-standing, piezoresistive microcantilever biosensor in a thermally-controlled system. The measured surface stresses showed a profile over a concentration range of 50–500 μg·mL−1, which covered the clinically therapeutic range of 50–100 μg·mL−1. The estimated limit of detection (LOD was calculated to be 45 μg·mL−1, and the binding affinity between the drug and the antibody was measured at around 90 ± 21 μg·mL−1. Lastly, the results of the proposed device showed a similar profile in valproic acid drug detection with those of the clinically-used fluorescence polarization immunoassay.

  4. Label-Free Biomedical Imaging Using High-Speed Lock-In Pixel Sensor for Stimulated Raman Scattering.

    Science.gov (United States)

    Mars, Kamel; Lioe, De Xing; Kawahito, Shoji; Yasutomi, Keita; Kagawa, Keiichiro; Yamada, Takahiro; Hashimoto, Mamoru

    2017-11-09

    Raman imaging eliminates the need for staining procedures, providing label-free imaging to study biological samples. Recent developments in stimulated Raman scattering (SRS) have achieved fast acquisition speed and hyperspectral imaging. However, there has been a problem of lack of detectors suitable for MHz modulation rate parallel detection, detecting multiple small SRS signals while eliminating extremely strong offset due to direct laser light. In this paper, we present a complementary metal-oxide semiconductor (CMOS) image sensor using high-speed lock-in pixels for stimulated Raman scattering that is capable of obtaining the difference of Stokes-on and Stokes-off signal at modulation frequency of 20 MHz in the pixel before reading out. The generated small SRS signal is extracted and amplified in a pixel using a high-speed and large area lateral electric field charge modulator (LEFM) employing two-step ion implantation and an in-pixel pair of low-pass filter, a sample and hold circuit and a switched capacitor integrator using a fully differential amplifier. A prototype chip is fabricated using 0.11 μm CMOS image sensor technology process. SRS spectra and images of stearic acid and 3T3-L1 samples are successfully obtained. The outcomes suggest that hyperspectral and multi-focus SRS imaging at video rate is viable after slight modifications to the pixel architecture and the acquisition system.

  5. Label-Free Biomedical Imaging Using High-Speed Lock-In Pixel Sensor for Stimulated Raman Scattering

    Directory of Open Access Journals (Sweden)

    Kamel Mars

    2017-11-01

    Full Text Available Raman imaging eliminates the need for staining procedures, providing label-free imaging to study biological samples. Recent developments in stimulated Raman scattering (SRS have achieved fast acquisition speed and hyperspectral imaging. However, there has been a problem of lack of detectors suitable for MHz modulation rate parallel detection, detecting multiple small SRS signals while eliminating extremely strong offset due to direct laser light. In this paper, we present a complementary metal-oxide semiconductor (CMOS image sensor using high-speed lock-in pixels for stimulated Raman scattering that is capable of obtaining the difference of Stokes-on and Stokes-off signal at modulation frequency of 20 MHz in the pixel before reading out. The generated small SRS signal is extracted and amplified in a pixel using a high-speed and large area lateral electric field charge modulator (LEFM employing two-step ion implantation and an in-pixel pair of low-pass filter, a sample and hold circuit and a switched capacitor integrator using a fully differential amplifier. A prototype chip is fabricated using 0.11 μm CMOS image sensor technology process. SRS spectra and images of stearic acid and 3T3-L1 samples are successfully obtained. The outcomes suggest that hyperspectral and multi-focus SRS imaging at video rate is viable after slight modifications to the pixel architecture and the acquisition system.

  6. Comparison of a Label-Free Quantitative Proteomic Method Based on Peptide Ion Current Area to the Isotope Coded Affinity Tag Method

    Directory of Open Access Journals (Sweden)

    Young Ah Goo

    2008-01-01

    Full Text Available Recently, several research groups have published methods for the determination of proteomic expression profiling by mass spectrometry without the use of exogenously added stable isotopes or stable isotope dilution theory. These so-called label-free, methods have the advantage of allowing data on each sample to be acquired independently from all other samples to which they can later be compared in silico for the purpose of measuring changes in protein expression between various biological states. We developed label free software based on direct measurement of peptide ion current area (PICA and compared it to two other methods, a simpler label free method known as spectral counting and the isotope coded affinity tag (ICAT method. Data analysis by these methods of a standard mixture containing proteins of known, but varying, concentrations showed that they performed similarly with a mean squared error of 0.09. Additionally, complex bacterial protein mixtures spiked with known concentrations of standard proteins were analyzed using the PICA label-free method. These results indicated that the PICA method detected all levels of standard spiked proteins at the 90% confidence level in this complex biological sample. This finding confirms that label-free methods, based on direct measurement of the area under a single ion current trace, performed as well as the standard ICAT method. Given the fact that the label-free methods provide ease in experimental design well beyond pair-wise comparison, label-free methods such as our PICA method are well suited for proteomic expression profiling of large numbers of samples as is needed in clinical analysis.

  7. Comparison of a label-free quantitative proteomic method based on peptide ion current area to the isotope coded affinity tag method.

    Science.gov (United States)

    Ryu, Soyoung; Gallis, Byron; Goo, Young Ah; Shaffer, Scott A; Radulovic, Dragan; Goodlett, David R

    2008-01-01

    Recently, several research groups have published methods for the determination of proteomic expression profiling by mass spectrometry without the use of exogenously added stable isotopes or stable isotope dilution theory. These so-called label-free, methods have the advantage of allowing data on each sample to be acquired independently from all other samples to which they can later be compared in silico for the purpose of measuring changes in protein expression between various biological states. We developed label free software based on direct measurement of peptide ion current area (PICA) and compared it to two other methods, a simpler label free method known as spectral counting and the isotope coded affinity tag (ICAT) method. Data analysis by these methods of a standard mixture containing proteins of known, but varying, concentrations showed that they performed similarly with a mean squared error of 0.09. Additionally, complex bacterial protein mixtures spiked with known concentrations of standard proteins were analyzed using the PICA label-free method. These results indicated that the PICA method detected all levels of standard spiked proteins at the 90% confidence level in this complex biological sample. This finding confirms that label-free methods, based on direct measurement of the area under a single ion current trace, performed as well as the standard ICAT method. Given the fact that the label-free methods provide ease in experimental design well beyond pair-wise comparison, label-free methods such as our PICA method are well suited for proteomic expression profiling of large numbers of samples as is needed in clinical analysis.

  8. Plasmonic biosensor for label-free G-quadruplexes detection

    Science.gov (United States)

    Qiu, Suyan; Zhao, Fusheng; Santos, Greggy M.; Shih, Wei-Chuan

    2016-03-01

    G-quadruplex, readily formed by the G-rich sequence, potentially distributes in over 40 % of all human genes, such as the telomeric DNA with the G-rich sequence found at the end of the chromosome. The G-quadruplex structure is supposed to possess a diverse set of critical functions in the mammalian genome for transcriptional regulation, DNA replication and genome stability. However, most of the currently available methods for G-quadruplex identification are restricted to fluorescence techniques susceptible to poor sensitivity. It is essential to propose methods with higher sensitivity to specifically recognize the G-quadruplexes. In this study, we demonstrate a label-free plasmonic biosensor for G-quadruplex detection by relying on the advantages of nanoporous gold (NPG) disks that provide high-density plasmonic hot spots, suitable for molecular recognition capability without the requirement for labeling processes.

  9. Nanophotonic label-free biosensors for environmental monitoring.

    Science.gov (United States)

    Chocarro-Ruiz, Blanca; Fernández-Gavela, Adrián; Herranz, Sonia; Lechuga, Laura M

    2017-06-01

    The field of environmental monitoring has experienced a substantial progress in the last years but still the on-site control of contaminants is an elusive problem. In addition, the growing number of pollutant sources is accompanied by an increasing need of having efficient early warning systems. Several years ago biosensor devices emerged as promising environmental monitoring tools, but their level of miniaturization and their fully operation outside the laboratory prevented their use on-site. In the last period, nanophotonic biosensors based on evanescent sensing have emerged as an outstanding choice for portable point-of-care diagnosis thanks to their capability, among others, of miniaturization, multiplexing, label-free detection and integration in lab-on-chip platforms. This review covers the most relevant nanophotonic biosensors which have been proposed (including interferometric waveguides, grating-couplers, microcavity resonators, photonic crystals and localized surface plasmon resonance sensors) and their recent application for environmental surveillance. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Label-free electrochemical genosensor based on mesoporous silica thin film.

    Science.gov (United States)

    Saadaoui, Maroua; Fernández, Iñigo; Luna, Gema; Díez, Paula; Campuzano, Susana; Raouafi, Noureddine; Sánchez, Alfredo; Pingarrón, José M; Villalonga, Reynaldo

    2016-10-01

    A novel label-free electrochemical strategy for nucleic acid detection was developed by using gold electrodes coated with mesoporous silica thin films as sensing interface. The biosensing approach relies on the covalent attachment of a capture DNA probe on the surface of the silica nanopores and further hybridization with its complementary target oligonucleotide sequence, causing a diffusion hindering of an Fe(CN)6 (3-/4-) electrochemical probe through the nanochannels of the mesoporous film. This DNA-mesoporous silica thin film-modified electrodes allowed sensitive (91.7 A/M) and rapid (45 min) detection of low nanomolar levels of synthetic target DNA (25 fmol) and were successfully employed to quantify the endogenous content of Escherichia coli 16S ribosomal RNA (rRNA) directly in raw bacterial lysate samples without isolation or purification steps. Moreover, the 1-month stability demonstrated by these biosensing devices enables their advanced preparation and storage, as desired for practical real-life applications. Graphical abstract Mesoporous silica thin films as scaffolds for the development of novel label-free electrochemical genosensors to perform selective, sensitive and rapid detection of target oligonucleotide sequences. Application towards E. coli determination.

  11. Label-free detection of C-reactive protein using an electrochemical DNA immunoassay

    Directory of Open Access Journals (Sweden)

    Temsiri Songjaroen

    2016-05-01

    Full Text Available A label-free electrochemical immunoassay that combines DNA-directed immobilization (DDI with electrochemical impedance spectroscopy (EIS on microwire sensors is reported for the detection of C-reactive protein (CRP. CRP is an acute-phase protein that is strongly correlated with systemic inflammation. Since inflammation plays a role in pathogenesis of cardiovascular diseases, CRP can be used to predict the likelihood of coronary events. To demonstrate the new chemistry, 25-μm Au electrodes were modified with single strand DNA (ssDNA and exposed to a solution containing complementary ssDNA conjugated to monoclonal anti-CRP. The charge-transfer resistance of the [Fe(CN6]3−/4− redox couple was used to determine the CRP concentration after binding. A stepwise increase in the charge transfer resistance was observed using EIS for each modification step, ssDNA, ssDNA-anti-CRP hybridization and the final CRP capture. Cyclic voltammetry (CV was used to verify the EIS results, and showed an increase in peak potential splitting in a similar stepwise manner for each modification step. Finally, fluorescence microscopy was used to confirm the DNA hybridization and CRP binding. Standard addition of CRP revealed that EIS could be used to detect CRP at clinically relevant levels in serum samples. This new form of electrochemical DNA immunoassay (eDI has significant potential as a simple, label-free sensor for proteins in microfluidic devices.

  12. Label free electrochemical aptasensor for ultrasensitive detection of ractopamine.

    Science.gov (United States)

    Yang, Fei; Wang, Peilong; Wang, Ruiguo; Zhou, Ying; Su, Xiaoou; He, Yujian; Shi, Lei; Yao, Dongsheng

    2016-03-15

    A label free electrochemical (EC) aptasensor for ultrasensitive detection of ractopamine (RAC) was developed. A special immobilization media consisting of gold nanoparticles/poly dimethyl diallyl ammonium chloride-graphene composite (AuNPs/PDDA-GN) was utilized to improve conductivity and performance of the biosensor. The RAC aptamer was attached on AuNPs of the composite membrane via Au-S bond. The fabrication process of the EC aptasensor was characterized by electrochemical impedance spectroscopy and cyclic voltammetry. The peak currents obtained by differential pulse voltammetry decreased linearly with the increasing of RAC concentrations and the sensor responds approximately logarithmically over a wide dynamic range of RAC concentration from 1.0 × 10(-12)mol/L to 1.0 × 10(-8)mol/L. The linear correlation coefficient of the developed aptasensor was 0.998, the limit of detection was 5.0 × 10(-13)mol/L. The proposed EC aptasensor displayed good stability, reproducibility and robust operation in animal urine. Particularly, the generality of the fabrication approach of electrochemical aptasensor is highlighted with a further example for illegal drugs detection via the aptamer identification. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Label-free high-throughput imaging flow cytometry

    Science.gov (United States)

    Mahjoubfar, A.; Chen, C.; Niazi, K. R.; Rabizadeh, S.; Jalali, B.

    2014-03-01

    Flow cytometry is an optical method for studying cells based on their individual physical and chemical characteristics. It is widely used in clinical diagnosis, medical research, and biotechnology for analysis of blood cells and other cells in suspension. Conventional flow cytometers aim a laser beam at a stream of cells and measure the elastic scattering of light at forward and side angles. They also perform single-point measurements of fluorescent emissions from labeled cells. However, many reagents used in cell labeling reduce cellular viability or change the behavior of the target cells through the activation of undesired cellular processes or inhibition of normal cellular activity. Therefore, labeled cells are not completely representative of their unaltered form nor are they fully reliable for downstream studies. To remove the requirement of cell labeling in flow cytometry, while still meeting the classification sensitivity and specificity goals, measurement of additional biophysical parameters is essential. Here, we introduce an interferometric imaging flow cytometer based on the world's fastest continuous-time camera. Our system simultaneously measures cellular size, scattering, and protein concentration as supplementary biophysical parameters for label-free cell classification. It exploits the wide bandwidth of ultrafast laser pulses to perform blur-free quantitative phase and intensity imaging at flow speeds as high as 10 meters per second and achieves nanometer-scale optical path length resolution for precise measurements of cellular protein concentration.

  14. Label-Free Biosensor Imaging on Photonic Crystal Surfaces.

    Science.gov (United States)

    Zhuo, Yue; Cunningham, Brian T

    2015-08-28

    We review the development and application of nanostructured photonic crystal surfaces and a hyperspectral reflectance imaging detection instrument which, when used together, represent a new form of optical microscopy that enables label-free, quantitative, and kinetic monitoring of biomaterial interaction with substrate surfaces. Photonic Crystal Enhanced Microscopy (PCEM) has been used to detect broad classes of materials which include dielectric nanoparticles, metal plasmonic nanoparticles, biomolecular layers, and live cells. Because PCEM does not require cytotoxic stains or photobleachable fluorescent dyes, it is especially useful for monitoring the long-term interactions of cells with extracellular matrix surfaces. PCEM is only sensitive to the attachment of cell components within ~200 nm of the photonic crystal surface, which may correspond to the region of most interest for adhesion processes that involve stem cell differentiation, chemotaxis, and metastasis. PCEM has also demonstrated sufficient sensitivity for sensing nanoparticle contrast agents that are roughly the same size as protein molecules, which may enable applications in "digital" diagnostics with single molecule sensing resolution. We will review PCEM's development history, operating principles, nanostructure design, and imaging modalities that enable tracking of optical scatterers, emitters, absorbers, and centers of dielectric permittivity.

  15. Label-free tissue scanner for colorectal cancer screening

    Science.gov (United States)

    Kandel, Mikhail E.; Sridharan, Shamira; Liang, Jon; Luo, Zelun; Han, Kevin; Macias, Virgilia; Shah, Anish; Patel, Roshan; Tangella, Krishnarao; Kajdacsy-Balla, Andre; Guzman, Grace; Popescu, Gabriel

    2017-06-01

    The current practice of surgical pathology relies on external contrast agents to reveal tissue architecture, which is then qualitatively examined by a trained pathologist. The diagnosis is based on the comparison with standardized empirical, qualitative assessments of limited objectivity. We propose an approach to pathology based on interferometric imaging of "unstained" biopsies, which provides unique capabilities for quantitative diagnosis and automation. We developed a label-free tissue scanner based on "quantitative phase imaging," which maps out optical path length at each point in the field of view and, thus, yields images that are sensitive to the "nanoscale" tissue architecture. Unlike analysis of stained tissue, which is qualitative in nature and affected by color balance, staining strength and imaging conditions, optical path length measurements are intrinsically quantitative, i.e., images can be compared across different instruments and clinical sites. These critical features allow us to automate the diagnosis process. We paired our interferometric optical system with highly parallelized, dedicated software algorithms for data acquisition, allowing us to image at a throughput comparable to that of commercial tissue scanners while maintaining the nanoscale sensitivity to morphology. Based on the measured phase information, we implemented software tools for autofocusing during imaging, as well as image archiving and data access. To illustrate the potential of our technology for large volume pathology screening, we established an "intrinsic marker" for colorectal disease that detects tissue with dysplasia or colorectal cancer and flags specific areas for further examination, potentially improving the efficiency of existing pathology workflows.

  16. Label-free analysis of physiological hyaluronan size distribution with a solid-state nanopore sensor.

    Science.gov (United States)

    Rivas, Felipe; Zahid, Osama K; Reesink, Heidi L; Peal, Bridgette T; Nixon, Alan J; DeAngelis, Paul L; Skardal, Aleksander; Rahbar, Elaheh; Hall, Adam R

    2018-03-12

    Hyaluronan (or hyaluronic acid, HA) is a ubiquitous molecule that plays critical roles in numerous physiological functions in vivo, including tissue hydration, inflammation, and joint lubrication. Both the abundance and size distribution of HA in biological fluids are recognized as robust indicators of various pathologies and disease progressions. However, such analyses remain challenging because conventional methods are not sufficiently sensitive, have limited dynamic range, and/or are only semi-quantitative. Here we demonstrate label-free detection and molecular weight discrimination of HA with a solid-state nanopore sensor. We first employ synthetic HA polymers to validate the measurement approach and then use the platform to determine the size distribution of as little as 10 ng of HA extracted directly from synovial fluid in an equine model of osteoarthritis. Our results establish a quantitative method for assessment of a significant molecular biomarker that bridges a gap in the current state of the art.

  17. Improved Label-Free LC-MS Analysis by Wavelet-Based Noise Rejection

    Directory of Open Access Journals (Sweden)

    Salvatore Cappadona

    2010-01-01

    Full Text Available Label-free LC-MS analysis allows determining the differential expression level of proteins in multiple samples, without the use of stable isotopes. This technique is based on the direct comparison of multiple runs, obtained by continuous detection in MS mode. Only differentially expressed peptides are selected for further fragmentation, thus avoiding the bias toward abundant peptides typical of data-dependent tandem MS. The computational framework includes detection, alignment, normalization and matching of peaks across multiple sets, and several software packages are available to address these processing steps. Yet, more care should be taken to improve the quality of the LC-MS maps entering the pipeline, as this parameter severely affects the results of all downstream analyses. In this paper we show how the inclusion of a preprocessing step of background subtraction in a common laboratory pipeline can lead to an enhanced inclusion list of peptides selected for fragmentation and consequently to better protein identification.

  18. Analytical technique for label-free multi-protein detection based on Western blot and surface-enhanced Raman scattering.

    Science.gov (United States)

    Han, Xiao X; Jia, Hui Y; Wang, Yan F; Lu, Zhi C; Wang, Chun X; Xu, Wei Q; Zhao, Bing; Ozaki, Yukihiro

    2008-04-15

    We have developed a new analytical procedure for label-free protein detection designated "Western SERS", consisting of protein electrophoresis, Western blot, colloidal silver staining, and surface-enhanced Raman scattering (SERS) detection. A novel method of silver staining for Western blot that uses a silver colloid, an excellent SERS-active substrate, is first proposed in the present study. During the process of silver staining, interactions between proteins and silver nanoparticles result in the emergence of SERS of proteins. In the present study, we use myoglobin (Mb) and bovine serum albumin (BSA) as model proteins. From different protein bands on a nitrocellulose (NC) membrane, we have observed surface-enhanced resonance Raman scattering (SERRS) spectra of Mb and SERS spectra of BSA. The proposed technique offers dual advantages of simplicity and high sensitivity. On one hand, after the colloidal silver staining, we can detect label-free multi-proteins directly on a NC membrane without digestion, extraction, and other pretreatments. On the other hand, the detection limit of the Western SERS is almost consistent with the detection limit of colloidal silver staining, and the SERRS detection limit of Mb is found to be 4 ng/band. This analytical method, which combines the technique of protein separation with SERS, may be a powerful protocol for label-free protein detection in proteomic research.

  19. Label-Free Electrochemical Immunoassay for C-Reactive Protein

    Directory of Open Access Journals (Sweden)

    Madasamy Thangamuthu

    2018-03-01

    Full Text Available C-reactive protein (CRP is one of the most expressed proteins in blood during acute phase inflammation, and its minute level increase has also been recognized for the clinical diagnosis of cardio vascular diseases. Unfortunately, the available commercial immunoassays are labour intensive, require large sample volumes, and have practical limitations, such as low stability and high production costs. Hence, we have developed a simple, cost effective, and label-free electrochemical immunoassay for the measurement of CRP in a drop of serum sample using an immunosensor strip made up of a screen printed carbon electrode (SPE modified with anti-CRP functionalized gold nanoparticles (AuNPs. The measurement relies on the decrease of the oxidation current of the redox indicator Fe3+/Fe2+, resulting from the immunoreaction between CRP and anti-CRP. Under optimal conditions, the present immunoassay measures CRP in a linear range from 0.4–200 nM (0.047–23.6 µg mL−1, with a detection limit of 0.15 nM (17 ng mL−1, S/N = 3 and sensitivity of 90.7 nA nM−1, in addition to a good reproducibility and storage stability. The analytical applicability of the presented immunoassay is verified by CRP measurements in human blood serum samples. This work provides the basis for a low-priced, safe, and easy-to-use point-of-care immunosensor assay to measure CRP at clinically relevant concentrations.

  20. Label-free probing of genes by time-domain terahertz sensing

    Energy Technology Data Exchange (ETDEWEB)

    Bolivar, P Haring [Institut fuer Halbleitertechnik, RWTH Aachen, Sommerfeldstr. 24, D-52056 Aachen (Germany); Brucherseifer, M [Institut fuer Halbleitertechnik, RWTH Aachen, Sommerfeldstr. 24, D-52056 Aachen (Germany); Nagel, M [Institut fuer Halbleitertechnik, RWTH Aachen, Sommerfeldstr. 24, D-52056 Aachen (Germany); Kurz, H [Institut fuer Halbleitertechnik, RWTH Aachen, Sommerfeldstr. 24, D-52056 Aachen (Germany); Bosserhoff, A [Institut fuer Pathologie, Universitaet Regensburg, Franz-Josef-Strauss-Allee 11, D-93053 Regensburg (Germany); Buettner, R [Institut fuer Pathologie, Universitaetsklinikum Bonn, Sigmund-Freud-Str. 25, D-53127 Bonn (Germany)

    2002-11-07

    A label-free sensing approach for the label-free characterization of genetic material with terahertz (THz) electromagnetic waves is presented. Time-resolved THz analysis of polynucleotides demonstrates a strong dependence of the complex refractive index of DNA molecules in the THz frequency range on their hybridization state. By monitoring THz signals one can thus infer the binding state (hybridized or denatured) of oligo- and polynucleotides, enabling the label-free determination the genetic composition of unknown DNA sequences. A broadband experimental proof-of-principle in a free-space analytic configuration, as well as a higher-sensitivity approach using integrated THz sensors reaching femtomol detection levels and demonstrating the capability to detect single-base mutations, are presented. The potential application for next generation high-throughput label-free genetic analytic systems is discussed.

  1. Photonic Crystal Biosensor Chip for Label-Free Detection of Bacteria

    DEFF Research Database (Denmark)

    Kristensen, Martin; Krüger, Asger Christian; Groothoff, Nathaniel

    Narrow polarization-mixing resonances in planar photonic crystals are studied as candidate components for label-free refractive index sensors for detecting bacteria causing sepsis through the identification of DNA strands....

  2. Label-free screening of single biomolecules through resistive pulse sensing technology for precision medicine applications

    Science.gov (United States)

    Harrer, S.; Kim, S. C.; Schieber, C.; Kannam, S.; Gunn, N.; Moore, S.; Scott, D.; Bathgate, R.; Skafidas, S.; Wagner, J. M.

    2015-05-01

    Employing integrated nano- and microfluidic circuits for detecting and characterizing biological compounds through resistive pulse sensing technology is a vibrant area of research at the interface of biotechnology and nanotechnology. Resistive pulse sensing platforms can be customized to study virtually any particle of choice which can be threaded through a fluidic channel and enable label-free single-particle interrogation with the primary read-out signal being an electric current fingerprint. The ability to perform label-free molecular screening with single-molecule and even single binding site resolution makes resistive pulse sensing technology a powerful tool for analyzing the smallest units of biological systems and how they interact with each other on a molecular level. This task is at the core of experimental systems biology and in particular ‘omics research which in combination with next-generation DNA-sequencing and next-generation drug discovery and design forms the foundation of a novel disruptive medical paradigm commonly referred to as personalized medicine or precision medicine. DNA-sequencing has approached the 1000-Dollar-Genome milestone allowing for decoding a complete human genome with unmatched speed and at low cost. Increased sequencing efficiency yields massive amounts of genomic data. Analyzing this data in combination with medical and biometric health data eventually enables understanding the pathways from individual genes to physiological functions. Access to this information triggers fundamental questions for doctors and patients alike: what are the chances of an outbreak for a specific disease? Can individual risks be managed and if so how? Which drugs are available and how should they be applied? Could a new drug be tailored to an individual’s genetic predisposition fast and in an affordable way? In order to provide answers and real-life value to patients, the rapid evolvement of novel computing approaches for analyzing big data in

  3. Development of an enrofloxacin immunosensor based on label-free electrochemical impedance spectroscopy.

    Science.gov (United States)

    Wu, Ching-Chou; Lin, Chia-Hung; Wang, Way-Shyan

    2009-06-30

    Enrofloxacin is the most widespread antibiotic in the fluoroquinolone family. As such, the development of a rapid and sensitive method for the determination of trace amounts of enrofloxacin is an important issue in the health field. The interaction of the enrofloxacin antigen to a specific antibody (Ab) immobilized on an 11-mercapto-undecanoic acid-coated gold electrode was quantified by electrochemical impedance spectroscopy. Two equivalent circuits were separately used to interpret the obtained impedance spectra. These circuits included one resistor in series with one parallel circuit comprised of a resistor and a capacitor (1R//C), and one resistor in series with two parallel RC circuits (2R//C). The results indicate that the antigen-antibody reaction analyzed using the 1R//C circuit provided a more sensitive resistance increment against the enrofloxacin concentration than that of the 2R//C circuit. However, the 2R//C circuit provided a better fitting for impedance spectra, and therefore supplies more detailed results of the enrofloxacin-antibody interaction, causing the increase of electron transfer resistance selectively to the modified layer, and not the electrical double layer. The antibody-modified electrode allowed for analysis of the dynamic linear range of 1-1000 ng/ml enrofloxacin with a detection limit of 1 ng/ml. The reagentless and label-free impedimetric immunosensors provide a simple and sensitive detection method for the specific determination of enrofloxacin.

  4. Impedimetric Label-Free Immunosensor on Disposable Modified Screen-Printed Electrodes for Ochratoxin A

    Directory of Open Access Journals (Sweden)

    Francesca Malvano

    2016-06-01

    Full Text Available An impedimetric label-free immunosensor on disposable screen-printed carbon electrodes (SPCE for quantitative determination of Ochratoxin A (OTA has been developed. After modification of the SPCE surface with gold nanoparticles (AuNPs, the anti-OTA was immobilized on the working electrode through a cysteamine layer. After each coating step, the modified surfaces were characterized by cyclic voltammetry (CV and electrochemical impedance spectroscopy (EIS. The capacitance was chosen as the best parameter that describes the reproducible change in electrical properties of the electrode surface at different OTA concentrations and it was used to investigate the analytical parameters of the developed immunosensor. Under optimized conditions, the immunosensor showed a linear relationship between 0.3 and 20 ng/mL with a low detection limit of 0.25 ng/mL, making it suitable to control OTA content in many common food products. Lastly, the immunosensor was used to measure OTA in red wine samples and the results were compared with those registered with a competitive ELISA kit. The immunosensor was sensitive to OTA lower than 2 μg/kg, which represents the lower acceptable limit of OTA established by European legislation for common food products.

  5. Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy

    Directory of Open Access Journals (Sweden)

    Dionisia Ortiz-Aguayo

    2018-01-01

    Full Text Available This research develops a label-free aptamer biosensor (aptasensor based on graphite-epoxy composite electrodes (GECs for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN6]3−/[Fe(CN6]4− as redox probe. After recording the frequency response, values were fitted to its electric model using the principle of equivalent circuits. The aptasensor showed a linear response up to 5 µM for lysozyme and a limit of detection of 1.67 µM. The sensitivity of the established method was 0.090 µM−1 in relative charge transfer resistance values. The interference response by main proteins, such as bovine serum albumin and cytochrome c, has been also characterized. To finally verify the performance of the developed aptasensor, it was applied to wine analysis.

  6. Label-free, ultrahigh-speed, 3D observation of bidirectional and correlated intracellular cargo transport by coherent brightfield microscopy.

    Science.gov (United States)

    Huang, Yi-Fan; Zhuo, Guan-Yu; Chou, Chun-Yu; Lin, Cheng-Hao; Hsieh, Chia-Lung

    2017-05-18

    The investigation of intracellular transport at the molecular scale requires measurements at high spatial and temporal resolutions. We demonstrate the label-free, direct imaging and tracking of native cell vesicles in live cells at an ultrahigh spatiotemporal resolution. Using coherent brightfield (COBRI) microscopy, we monitor individual cell vesicles traveling inside the cell with nanometer spatial precision in 3D at 30 000 frames per second. The stepwise directional motion of the vesicle on the cytoskeletal track is clearly resolved. We also observe the repeated switching of the transport direction of the vesicle in a continuous trajectory. Our high-resolution measurement unveils the transient pausing and subtle bidirectional motion of the vesicle, taking place over tens of nanometers in tens of milliseconds. By tracking multiple particles simultaneously, we found strong correlations between the motions of two neighboring vesicles. Our label-free ultrahigh-speed optical imaging provides the opportunity to visualize intracellular cargo transport at the nanoscale in the microsecond timescale with minimal perturbation.

  7. Development of a label-free aptasensor for monitoring the self-association of lysozyme.

    Science.gov (United States)

    Vasilescu, Alina; Gaspar, Szilveszter; Mihai, Iuliana; Tache, Andreia; Litescu, Simona Carmen

    2013-06-21

    A novel aptamer and surface plasmon resonance (SPR)-based sensor was developed for the label-free detection of lysozyme. The aptasensor is characterised by a detection limit of 1 μg mL(-1) and a linear range of 5-50 μg mL(-1). As an application, we examined the usefulness of the aptasensor for monitoring the early stages of the aggregation of lysozyme. It was surprisingly found that, despite a significant decrease in monomer content during aggregation, the response of the aptasensor for protein solutions aged for 12 hours was similar to that for the fresh protein. To correlate the results obtained with the aptasensor with the composition of lysozyme solutions at various time points, we examined them in detail by atomic force microscopy (AFM), thioflavin T fluorescence, size-exclusion chromatography (SEC) and Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectrometry (MALDI-TOF-MS). All methods together indicated that during the initial hours of aggregation, the protein solutions contained small lysozyme oligomers (mainly dimers) and decreasing amounts of monomers. Our results thus suggest that the aptamer also recognizes lysozyme dimers/oligomers. A higher non-specific binding was observed for the aggregated lysozyme at the surface of the aptasensor as compared to the native protein. This was attributed to the hydrophobic patches which are exposed by the unfolded lysozyme and/or oligomer species, allowing for different adsorption and organisation at the surface of the aptasensor. This hypothesis is supported by square wave voltammetry (SWV) studies using solutions of aggregated lysozyme. A higher electrochemical signal due to the direct oxidation of tyrosine/tryptophan residues was observed for aged protein solutions as compared to the fresh solution, indicative of an increased number of such exposed electroactive residues and of overall increased surface hydrophobicity of the protein. Our work presents a label-free lysozyme aptasensor that is

  8. 78 FR 38195 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-06-26

    ... Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT... all General Electric Company (GE) GE90-110B1 and GE90-115B turbofan engines. This emergency AD was...) Applicability General Electric Company (GE) GE90-110B1 and GE90-115B turbofan engines with a transfer gearbox...

  9. 78 FR 72552 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-12-03

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... General Electric Company model GEnx-2B67 and GEnx-2B67B turbofan engines. This AD was prompted by the... certain serial number General Electric Company (GE) model GEnx-2B67 and GEnx-2B67B turbofan engines. The...

  10. Label-free investigation of the effects of lithium niobate polarization on cell adhesion

    Science.gov (United States)

    Mandracchia, B.; Gennari, O.; Paturzo, M.; Grilli, S.; Ferraro, P.

    2017-06-01

    The determination of contact area is pivotal to understand how biomaterials properties influence cell adhesion. In particular, the influence of surface charges is well-known but still controversial, especially when new functional materials and methods are introduced. Here, we use for the first time Holographic Total Internal Reflection Microscopy (HoloTIRM) to study the influence of the spontaneous polarization of ferroelectric lithium niobate (LN) on the adhesion properties of fibroblast cells. The selective illumination of a very thin region directly above the substrate, achieved by Total Internal Reflection, provides high-contrast images of the contact regions. Holographic recording, on the other hand, allows for label-free quantitative phase imaging of the contact areas between cells and LN. Phase signal is more sensitive in the first 100nm and, thus more reliable in order to locate focal contacts. This work shows that cells adhering on negatively polarized LN present a significant increase of the contact area in comparison with cells adhering on the positively polarized LN substrate, as well as an intensification of contact vicinity. This confirms the potential of LN as a platform for investigating the role of charges on cellular processes. The similarity of cell adhesion behavior on negatively polarized LN and glass control also confirms the possibility to use LN as an active substrate without impairing cell behavior.

  11. Label-free potentiometry for detecting DNA hybridization using peptide nucleic acid and DNA probes.

    Science.gov (United States)

    Goda, Tatsuro; Singi, Ankit Balram; Maeda, Yasuhiro; Matsumoto, Akira; Torimura, Masaki; Aoki, Hiroshi; Miyahara, Yuji

    2013-02-07

    Peptide nucleic acid (PNA) has outstanding affinity over DNA for complementary nucleic acid sequences by forming a PNA-DNA heterodimer upon hybridization via Watson-Crick base-pairing. To verify whether PNA probes on an electrode surface enhance sensitivity for potentiometric DNA detection or not, we conducted a comparative study on the hybridization of PNA and DNA probes on the surface of a 10-channel gold electrodes microarray. Changes in the charge density as a result of hybridization at the solution/electrode interface on the self-assembled monolayer (SAM)-formed microelectrodes were directly transformed into potentiometric signals using a high input impedance electrometer. The charge readout allows label-free, reagent-less, and multi-parallel detection of target oligonucleotides without any optical assistance. The differences in the probe lengths between 15- to 22-mer dramatically influenced on the sensitivity of the PNA and DNA sensors. Molecular type of the capturing probe did not affect the degree of potential shift. Theoretical model for charged rod-like duplex using the Gouy-Chapman equation indicates the dominant effect of electrostatic attractive forces between anionic DNA and underlying electrode at the electrolyte/electrode interface in the potentiometry.

  12. Label-Free Potentiometry for Detecting DNA Hybridization Using Peptide Nucleic Acid and DNA Probes

    Science.gov (United States)

    Goda, Tatsuro; Singi, Ankit Balram; Maeda, Yasuhiro; Matsumoto, Akira; Torimura, Masaki; Aoki, Hiroshi; Miyahara, Yuji

    2013-01-01

    Peptide nucleic acid (PNA) has outstanding affinity over DNA for complementary nucleic acid sequences by forming a PNA-DNA heterodimer upon hybridization via Watson-Crick base-pairing. To verify whether PNA probes on an electrode surface enhance sensitivity for potentiometric DNA detection or not, we conducted a comparative study on the hybridization of PNA and DNA probes on the surface of a 10-channel gold electrodes microarray. Changes in the charge density as a result of hybridization at the solution/electrode interface on the self-assembled monolayer (SAM)-formed microelectrodes were directly transformed into potentiometric signals using a high input impedance electrometer. The charge readout allows label-free, reagent-less, and multi-parallel detection of target oligonucleotides without any optical assistance. The differences in the probe lengths between 15- to 22-mer dramatically influenced on the sensitivity of the PNA and DNA sensors. Molecular type of the capturing probe did not affect the degree of potential shift. Theoretical model for charged rod-like duplex using the Gouy-Chapman equation indicates the dominant effect of electrostatic attractive forces between anionic DNA and underlying electrode at the electrolyte/electrode interface in the potentiometry. PMID:23435052

  13. Ultrasensitive, label-free detection of cardiac biomarkers with optical SIS sensor.

    Science.gov (United States)

    Diware, Mangesh S; Cho, Hyun Mo; Chegal, Won; Cho, Yong Jai; Kim, Dong Soo; O, Sang Won; Kim, Kyeong-Suk; Paek, Se-Hwan

    2017-01-15

    Acute myocardial infarction (MI) is the leading cause of high mortality and morbidity rate worldwide, early and accurate diagnosis can increase the chances of survival. In this work, we report a simple, ultrasensitive, label-free, and high-throughput solution immersed silicon (SIS) immunosensor based on non-reflection condition (NRC) for p-polarized wave for early diagnosis of MI. SIS sensor chips are just a thin dielectric polymer layer on the silicon surface, which can be functionalized for specific application. At NRC, SIS sensors are extremely sensitive to the growing thickness of a bio-layer on the sensor surface while independent of refractive index change of the surrounding medium. Therefore, SIS signal is free from thermal noise, unlike surface plasmon resonance based sensor. Also, there is no need of reference signal which facilitates fast and accurate interaction measurement. Here, SIS technology is applied to tackle two issues in MI diagnosis: high sensitivity with the direct assay and the ability to measure in human serum. Myoglobin, creatine kinase-MB, and cardiac troponin I (cTnI) proteins were used as the MI biomarkers. We were able to measure over a broad concentration range with the detection limit of 5 and 10pg/ml for cTnI in PBS and blood serum, respectively. The response time is about 5min. This novel technique is a suitable candidate for cost effective point-of-care application. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Label-free carbon particulates detection in bio (medical) settings (Conference Presentation)

    Science.gov (United States)

    Steuwe, Christian; Bové, Hannelore; vandeVen, Martin J.; Ameloot, Marcel; Roeffaers, Maarten B. J.

    2017-02-01

    The adverse health effects of particulate matter exposure are a generally accepted concern. Dramatic statistical figures suggest that fine dust is a main environmental risk in Europe and can be held accountable for hundreds of thousands of deaths per year [1]. Locating and tracking these nanometer sized particles, however, is not straight forward: In epidemiological and toxicology research only measurements based on labels [2] such as radionuclide markers have been applied. In this paper we present a direct, label-free optical contrast mechanism to detect carbon nanoparticles immersed in aqueous environments [3]. The virtue of this technique is its ability to perform in body fluids such as urine but also in cells and tissues. The mechanism is based on white light (WL) generation upon illumination with femtosecond pulsed near-infrared and is therefore non-incandescence related. We demonstrate the technique in various biological settings with dry and suspended carbon black particles (CB), a widely used model compound for soot [4]. Our approach allows for the unequivocal localization of CB alongside of common fluorophores and markers and can be performed on multiphoton laser-scanning microscopy platforms, a system commonly available in research laboratories. [1] European Environment Agency (2015). Press release. [2] Kong et al. Int. J. Mol. Sci. 2013, 14, (11), 22529-22543 [3] Bové and Steuwe et al. Nano letters, 2016, (16) , pages 3173-3178 [4] Arnal et al. Combust. Sci. Technol. 2012, 184, (7-8), 1191-1206.

  15. Label-free recognition of drug resistance via impedimetric screening of breast cancer cells.

    Directory of Open Access Journals (Sweden)

    Bilge Eker

    Full Text Available We present a novel study on label-free recognition and distinction of drug resistant breast cancer cells (MCF-7 DOX from their parental cells (MCF-7 WT via impedimetric measurements. Drug resistant cells exhibited significant differences in their dielectric properties compared to wild-type cells, exerting much higher extracellular resistance (Rextra . Immunostaining revealed that MCF-7 DOX cells gained a much denser F-actin network upon acquiring drug resistance indicating that remodeling of actin cytoskeleton is probably the reason behind higher Rextra , providing stronger cell architecture. Moreover, having exposed both cell types to doxorubicin, we were able to distinguish these two phenotypes based on their substantially different drug response. Interestingly, impedimetric measurements identified a concentration-dependent and reversible increase in cell stiffness in the presence of low non-lethal drug doses. Combined with a profound frequency analysis, these findings enabled distinguishing distinct cellular responses during drug exposure within four concentration ranges without using any labeling. Overall, this study highlights the possibility to differentiate drug resistant phenotypes from their parental cells and to assess their drug response by using microelectrodes, offering direct, real-time and noninvasive measurements of cell dependent parameters under drug exposure, hence providing a promising step for personalized medicine applications such as evaluation of the disease progress and optimization of the drug treatment of a patient during chemotherapy.

  16. Label-Free QCM Immunosensor for the Detection of Ochratoxin A.

    Science.gov (United States)

    Pirinçci, Şerife Şeyda; Ertekin, Özlem; Laguna, Duygu Ercan; Özen, Fehime Şeyma; Öztürk, Zafer Ziya; Öztürk, Selma

    2018-04-11

    Ochratoxin A (OTA) is a potent mycotoxin that poses a risk in food and feed moieties and subject to worldwide regulation. Laboratory-based analytical methods are traditionally employed for reliable OTA quantification, but these methods cannot provide rapid and on-site analysis, where biosensors fill this gap. In this study a label-free quartz crystal microbalance (QCM)-based immunosensor for the detection of OTA, which is one of the most important small molecule contaminants, was developed by direct immobilization of OTA to amine-bearing sensor surfaces using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-Hydroxysuccinimide (NHS) chemistry. The protein-free sensor surface enabled regeneration of sensor surface with 50 mM NaOH and 1% SDS up to 13 times without loss of performance, which would disrupt a protein-containing sensor surface. We developed a QCM immunosensor using the developed sensor surface with a 17.2-200 ng/mL detection range which can be used for on-site detection of feedstuffs.

  17. Direct Conversion of Radioisotope Energy to Electricity

    Energy Technology Data Exchange (ETDEWEB)

    Marks Prelas; Alexey Spitsyn; Alejandro Suarez; Eric Stienfelds; Dickerson Moreno; Bia-Ling Hsu; Tushar Ghosh; Robert Tompson; Sudarshan Loyalka; Dabir Viswanath

    2003-09-09

    A new chemical reactor has been tested for Field Enhanced Diffusion by Optical Activation doping and purification of SiC, GaN and AlN films. Different conditions have been used on SiC, GaN and AlN samples including temperature variation, electrical field variation, variations in electrical current and optical activation. A 5mW (630-680) nm laser was used for optical activation. It was observed that optical activation has a major effect on ion drift rates. It was also observed that the magnitude of the electrical current also enhanced ion drift rates by a postulated current drag mechanism. I-V characteristic curves were measured to verify changes in the electrical properties of the samples SIMS was used to analyze the concentrations of impurities in the film samples before and after treatment. It has been demonstrated that the field-enhanced diffusion by optical activation method can dope and purify the films. As a result, the electrical properties of the wafers have been significantly improved during treatment especially in cases where a laser is used.

  18. 77 FR 74125 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2012-12-13

    ...-1288; Directorate Identifier 2012-NE-37-AD] RIN 2120-AA64 Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of proposed rulemaking (NPRM). SUMMARY: We propose to adopt a new airworthiness directive (AD) for all General Electric...

  19. Extracting histones for the specific purpose of label-free MS.

    Science.gov (United States)

    Govaert, Elisabeth; Van Steendam, Katleen; Scheerlinck, Ellen; Vossaert, Liesbeth; Meert, Paulien; Stella, Martina; Willems, Sander; De Clerck, Laura; Dhaenens, Maarten; Deforce, Dieter

    2016-12-01

    Extracting histones from cells is the first step in studies that aim to characterize histones and their post-translational modifications (hPTMs) with MS. In the last decade, label-free quantification is more frequently being used for MS-based histone characterization. However, many histone extraction protocols were not specifically designed for label-free MS. While label-free quantification has its advantages, it is also very susceptible to technical variation. Here, we adjust an established histone extraction protocol according to general label-free MS guidelines with a specific focus on minimizing sample handling. These protocols are first evaluated using SDS-PAGE. Hereafter, a selection of extraction protocols was used in a complete histone workflow for label-free MS. All protocols display nearly identical relative quantification of hPTMs. We thus show that, depending on the cell type under investigation and at the cost of some additional contaminating proteins, minimizing sample handling can be done during histone isolation. This allows analyzing bigger sample batches, leads to reduced technical variation and minimizes the chance of in vitro alterations to the hPTM snapshot. Overall, these results allow researchers to determine the best protocol depending on the resources and goal of their specific study. Data are available via ProteomeXchange with identifier PXD002885. © 2016 The Authors. Proteomics Published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Progress of new label-free techniques for biosensors: a review.

    Science.gov (United States)

    Sang, Shengbo; Wang, Yajun; Feng, Qiliang; Wei, Ye; Ji, Jianlong; Zhang, Wendong

    2016-01-01

    The detection techniques used in biosensors can be broadly classified into label-based and label-free. Label-based detection relies on the specific properties of labels for detecting a particular target. In contrast, label-free detection is suitable for the target molecules that are not labeled or the screening of analytes which are not easy to tag. Also, more types of label-free biosensors have emerged with developments in biotechnology. The latest developed techniques in label-free biosensors, such as field-effect transistors-based biosensors including carbon nanotube field-effect transistor biosensors, graphene field-effect transistor biosensors and silicon nanowire field-effect transistor biosensors, magnetoelastic biosensors, optical-based biosensors, surface stress-based biosensors and other type of biosensors based on the nanotechnology are discussed. The sensing principles, configurations, sensing performance, applications, advantages and restriction of different label-free based biosensors are considered and discussed in this review. Most concepts included in this survey could certainly be applied to the development of this kind of biosensor in the future.

  1. Calculating Electrical Requirements for Direct Current Electric Actuators

    Science.gov (United States)

    2017-11-29

    windings and internal wiring. The inductance is created from the turns of the wire that make up the windings. The voltage source is a result of the back...as the energy charges the bus capacitance. If the voltage rises too high, it could exceed voltage ratings of components and cause damage. To...watts) Q Electric charge (coulombs)  Efficiency (ratio less than one) Η Corrected efficiency (represents  or 1/ depending on state) r Speed

  2. A low-cost, label-free DNA detection method in lab-on-chip format based on electrohydrodynamic instabilities, with application to long-range PCR.

    Science.gov (United States)

    Diakité, Mohamed Lemine Youba; Champ, Jerôme; Descroix, Stephanie; Malaquin, Laurent; Amblard, François; Viovy, Jean-Louis

    2012-11-21

    In order to evolve from a "chip in the lab" to a "lab on a chip" paradigm, there is still a strong demand for low-cost, portable detection technologies, notably for analytes at low concentrations. Here we report a new label-free DNA detection method with direct electronic read, and apply it to long-range PCR. This method uses a nonlinear electrohydrodynamic phenomenon: when subjected to high electric fields (typically above 100 V cm(-1)), suspensions of large polyelectrolytes, such as long DNA molecules, create "giant" dynamic concentration fluctuations. These fluctuations are associated with large conductivity inhomogeneities, and we use here a contact-mode local conductivity detector to detect these fluctuations. In order to decouple the detection electronics from the high voltage excitation one, an original "doubly symmetric" floating mode battery-operated detection scheme was developed. A wavelet analysis was then applied, to unravel from the chaotic character of the electohydrodynamic instabilities a scalar signal robustly reflecting the amplification of DNA. As a first proof of concept, we measured the products of the off-chip amplification of 10 kbp DNA from lambda phage DNA, achieving a sensitivity better than 100 fg DNA in the original 50 μl sample. This corresponds to the amplification products of less than 100 initial copies of target DNA. The companion enabling technologies developed to implement this new concept, i.e. the doubly symmetric contact conductivity detection and wavelet analysis, may also find various other applications in lab-on-chips.

  3. Label-free aptamer-based sensor for specific detection of malathion residues by surface-enhanced Raman scattering

    Science.gov (United States)

    Nie, Yonghui; Teng, Yuanjie; Li, Pan; Liu, Wenhan; Shi, Qianwei; Zhang, Yuchao

    2018-02-01

    A novel label-free aptamer surface-enhanced Raman scattering (SERS) sensor for trace malathion residue detection was proposed. In this process, the binding of malathion molecule with aptamer is identified directly. The silver nanoparticles modified with positively charged spermine served as enhancing and capture reagents for the negatively charged aptamer. Then, the silver nanoparticles modified by aptamer were used to specifically capture the malathion. The SERS background spectra of spermine, aptamer, and malathion were recorded and distinguished with the spectrum of malathion-aptamer. To enhance the characteristic peak signal of malathion captured by the aptamer, the aggregate reagents (NaCl, KCl, MgCl2) were compared and selected. The selectivity of this method was verified in the mixed-pesticide standard solution, which included malathion, phosmet, chlorpyrifos-methyl, and fethion. Results show that malathion can be specifically identified when the mixed-pesticide interferences existed. The standard curve was established, presenting a good linear range of 5 × 10- 7 to 1 × 10- 5 mol·L- 1. The spiked experiments for tap water show good recoveries from 87.4% to 110.5% with a relative standard deviation of less than 4.22%. Therefore, the proposed label-free aptamer SERS sensor is convenient, specifically detects trace malathion residues, and can be applied for qualitative and quantitative analysis of other pesticides.

  4. Label and label-free based surface-enhanced Raman scattering for pathogen bacteria detection: A review.

    Science.gov (United States)

    Liu, Yu; Zhou, Haibo; Hu, Ziwei; Yu, Guangxia; Yang, Danting; Zhao, Jinshun

    2017-08-15

    Rapid, accurate detection of pathogen bacteria is a highly topical research area for the sake of food safety and public health. Surface-enhanced Raman scattering (SERS) is being considered as a powerful and attractive technique for pathogen bacteria detection, due to its sensitivity, high speed, comparatively low cost, multiplexing ability and portability. This contribution aims to give a comprehensive overview of SERS as a technique for rapid detection of pathogen bacteria based on label and label-free strategies. A brief tutorial on SERS is given first of all. Then we summarize the recent trends and developments of label and label-free based SERS applied to detection of pathogen bacteria, including the relatively complete interpretation of SERS spectra. In addition, multifunctional SERS platforms for pathogen bacteria in matrix are discussed as well. Furthermore, an outlook of the work done and a perspective on the future directions of SERS as a reliable tool for real-time pathogen bacteria detection are given. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. A SIMULTANEOUS MULTI-PROBE DETECTION LABEL-FREE OPTICAL-RESOLUTION PHOTOACOUSTIC MICROSCOPY TECHNIQUE BASED ON MICROCAVITY TRANSDUCER

    Directory of Open Access Journals (Sweden)

    YONGBO WU

    2013-07-01

    Full Text Available We demonstrate the feasibility of simultaneous multi-probe detection for an optical-resolution photoacoustic microscopy (OR-PAM system. OR-PAM has elicited the attention of biomedical imaging researchers because of its optical absorption contrast and high spatial resolution with great imaging depth. OR-PAM allows label-free and noninvasive imaging by maximizing the optical absorption of endogenous biomolecules. However, given the inadequate absorption of some biomolecules, detection sensitivity at the same incident intensity requires improvement. In this study, a modulated continuous wave with power density less than 3 mW/cm2 (1/4 of the ANSI safety limit excited the weak photoacoustic (PA signals of biological cells. A microcavity transducer is developed based on the bulk modulus of gas five orders of magnitude lower than that of solid; air pressure variation is inversely proportional to cavity volume at the same temperature increase. Considering that a PA wave expands in various directions, detecting PA signals from different positions and adding them together can increase detection sensitivity and signal-to-noise ratio. Therefore, we employ four detectors to acquire tiny PA signals simultaneously. Experimental results show that the developed OR-PAM system allows the label-free imaging of cells with weak optical absorption.

  6. Label-free SRM-based relative quantification of antibiotic resistance mechanisms in Pseudomonas aeruginosa clinical isolates

    Directory of Open Access Journals (Sweden)

    Yannick eCharretier

    2015-02-01

    Full Text Available Both acquired and intrinsic mechanisms play a crucial role in Pseudomonas aeruginosa antibiotic resistance. Many clinically relevant resistance mechanisms result from changes in gene expression, namely multidrug efflux pump overproduction, AmpC beta-lactamase induction or derepression, and inactivation or repression of the carbapenem-specific porin OprD. Changes in gene expression are usually assessed using reverse-transcription quantitative real-time PCR (RT-qPCR assays. Here, we evaluated label-free Selected Reaction Monitoring (SRM-based mass spectrometry to directly quantify proteins involved in antibiotic resistance. We evaluated the label-free SRM using a defined set of P. aeruginosa isolates with known resistance mechanisms and compared it against RT-qPCR. Referring to efflux systems, we found a more robust relative quantification of antibiotic resistance mechanisms by SRM than RT-qPCR. The SRM-based approach was applied to a set of clinical P. aeruginosa isolates to detect antibiotic resistance proteins. This multiplexed SRM-based approach is a rapid and reliable method for the simultaneous detection and quantification of resistance mechanisms and we demonstrate its relevance for antibiotic resistance prediction.

  7. Label-free SRM-based relative quantification of antibiotic resistance mechanisms in Pseudomonas aeruginosa clinical isolates.

    Science.gov (United States)

    Charretier, Yannick; Köhler, Thilo; Cecchini, Tiphaine; Bardet, Chloé; Cherkaoui, Abdessalam; Llanes, Catherine; Bogaerts, Pierre; Chatellier, Sonia; Charrier, Jean-Philippe; Schrenzel, Jacques

    2015-01-01

    Both acquired and intrinsic mechanisms play a crucial role in Pseudomonas aeruginosa antibiotic resistance. Many clinically relevant resistance mechanisms result from changes in gene expression, namely multidrug efflux pump overproduction, AmpC β-lactamase induction or derepression, and inactivation or repression of the carbapenem-specific porin OprD. Changes in gene expression are usually assessed using reverse-transcription quantitative real-time PCR (RT-qPCR) assays. Here, we evaluated label-free Selected Reaction Monitoring (SRM)-based mass spectrometry to directly quantify proteins involved in antibiotic resistance. We evaluated the label-free SRM using a defined set of P. aeruginosa isolates with known resistance mechanisms and compared it with RT-qPCR. Referring to efflux systems, we found a more robust relative quantification of antibiotic resistance mechanisms by SRM than RT-qPCR. The SRM-based approach was applied to a set of clinical P. aeruginosa isolates to detect antibiotic resistance proteins. This multiplexed SRM-based approach is a rapid and reliable method for the simultaneous detection and quantification of resistance mechanisms and we demonstrate its relevance for antibiotic resistance prediction.

  8. Label-free measurements of reaction kinetics using a droplet-based optofluidic device.

    Science.gov (United States)

    Mao, Zhangming; Guo, Feng; Xie, Yuliang; Zhao, Yanhui; Lapsley, Michael Ian; Wang, Lin; Mai, John D; Costanzo, Francesco; Huang, Tony Jun

    2015-02-01

    Label-free measurements of the reaction kinetics of a small sample volume are essential for efficient drug discovery, requiring methods and systems that are rapid, accurate, and cost-effective. Herein, we present an integrated optofluidic system for label-free characterization of reactions in a nanoliter reagent volume. This system contains a droplet-based microfluidic sampling section and an optical fiber-based spectroscopy detection section. By manipulating droplets containing reagents at certain concentrations at different times, quantifiable measurements via absorption spectroscopy can be made in a simple, sensitive, and high-throughput manner. We have demonstrated our system's capability by performing potency (IC50) assays of an inhibitor in a TEM-1 β-lactamase (enzyme) and nitrocefin (substrate) system. This integrated platform can potentially provide an automated, label-free, and low-cost method for many other assays of reaction kinetics. © 2014 Society for Laboratory Automation and Screening.

  9. An adaptive alignment algorithm for quality-controlled label-free LC-MS.

    Science.gov (United States)

    Sandin, Marianne; Ali, Ashfaq; Hansson, Karin; Månsson, Olle; Andreasson, Erik; Resjö, Svante; Levander, Fredrik

    2013-05-01

    Label-free quantification using precursor-based intensities is a versatile workflow for large-scale proteomics studies. The method however requires extensive computational analysis and is therefore in need of robust quality control during the data mining stage. We present a new label-free data analysis workflow integrated into a multiuser software platform. A novel adaptive alignment algorithm has been developed to minimize the possible systematic bias introduced into the analysis. Parameters are estimated on the fly from the data at hand, producing a user-friendly analysis suite. Quality metrics are output in every step of the analysis as well as actively incorporated into the parameter estimation. We furthermore show the improvement of this system by comprehensive comparison to classical label-free analysis methodology as well as current state-of-the-art software.

  10. Atomic Force Microscopy Images Label-Free, Drug Encapsulated Nanoparticles In Vivo and Detects Difference in Tissue Mechanical Properties of Treated and Untreated: A Tip for Nanotoxicology

    Science.gov (United States)

    Lamprou, Dimitrios A.; Venkatpurwar, Vinod; Kumar, M. N. V. Ravi

    2013-01-01

    Overcoming the intractable challenge of imaging of label-free, drug encapsulated nanoparticles in tissues in vivo would directly address associated regulatory concerns over 'nanotoxicology'. Here we demonstrate the utility of Atomic Force Microscopy (AFM) for visualising label-free, drug encapsulated polyester particles of ∼280 nm distributed within tissues following their intravenous or peroral administration to rodents. A surprising phenomenon, in which the tissues' mechanical stiffness was directly measured (also by AFM) and related to the number of embedded nanoparticles, was utilised to generate quantitative data sets for nanoparticles localisation. By coupling the normal determination of a drug's pharmacokinetics/pharmacodynamics with post-sacrifice measurement of nanoparticle localisation and number, we present for the first time an experimental design in which a single in vivo study relates the PK/PD of a nanomedicine to its toxicokinetics. PMID:23724054

  11. iTRAQ-Based and Label-Free Proteomics Approaches for Studies of Human Adenovirus Infections

    OpenAIRE

    Trinh, Hung V.; Grossmann, Jonas; Gehrig, Peter; Roschitzki, Bernd; Schlapbach, Ralph; Greber, Urs F.; Hemmi, Silvio

    2013-01-01

    Both isobaric tags for relative and absolute quantitation (iTRAQ) and label-free methods are widely used for quantitative proteomics. Here, we provide a detailed evaluation of these proteomics approaches based on large datasets from biological samples. iTRAQ-label-based and label-free quantitations were compared using protein lysate samples from noninfected human lung epithelial A549 cells and from cells infected for 24 h with human adenovirus type 3 or type 5. Either iTRAQ-label-based or lab...

  12. High Torque, Direct Drive Electric Motor Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Bear Engineering proposes to advance the development of an innovative high torque, low speed, direct drive motor in order to meet NASA's requirements for such...

  13. New directions in electric power financing

    International Nuclear Information System (INIS)

    Jechoutek, K.G.; Lamech, Ranjit

    1995-01-01

    This paper argues that it is necessary to raise the eyes from the current focus on independent power projects, buttressed by guarantees, to the longer horizon of electric power financing in open markets. Transitional strategies will need to move beyond the commonly seen IPP activity that occurs without fundamental sector reform, and demand-side incentives that introduce further market distortions. These efforts will have to focus on macroeconomic stabilization, removal of price distortions, as well as sector and corporate reform. Mobilization of domestic capital will be essential for sustainable sector financing. Although guarantees to encourage power sector investment can be designed to selectively cover risks, their elimination through fundamental sector reform should be the ultimate goal. Over the longer-term traditional corporate finance should become a more common financing strategy than project finance. Innovations in performance risk management and consumer credit will be crucial to the financing of energy efficiency. (author)

  14. 78 FR 72567 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-12-03

    ...) Definition For the purpose of this AD, a shop visit is the induction of an engine into the shop for... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... General Electric Company (GE) GE90-110B1 and -115B turbofan engines. This AD was prompted by multiple...

  15. 77 FR 3088 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2012-01-23

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... Electric Company (GE) CF34-10E series turbofan engines. This AD was prompted by a report of heavy wear... removing from service all CVD support assemblies and any fan drive shaft on the affected engines if wear is...

  16. 78 FR 19628 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-04-02

    ... Electric Company (GE) GE90-76B, -85B, -90B, -94B, - 110B1, and -115B turbofan engines. This proposed AD was...-0186; Directorate Identifier 2013-NE-11-AD] RIN 2120-AA64 Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of proposed...

  17. 78 FR 56594 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-09-13

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... General Electric Company (GE) GE90-76B, -85B, -90B, -94B, -110B1, and - 115B turbofan engines. This AD was... to accelerated corrosion and oxidation, including one engine in- flight shutdown (IFSD) caused by...

  18. 78 FR 76045 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-12-16

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... (AD) for General Electric Company (GE) GE90-110B1 and GE90-115B turbofan engines with certain high... rotating engine part, which could result in an uncontained engine failure and damage to the airplane. DATES...

  19. 78 FR 19983 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-04-03

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... Electric Company (GE) CF34-8C and CF34-8E turbofan engines with certain part numbers (P/N) of operability... the OBV. Two of those events led to an engine fire. This AD requires the affected OBVs be removed from...

  20. Multiplex surface plasmon resonance imaging platform for label-free detection of foodborne pathogens

    Science.gov (United States)

    Salmonellae are among the leading causes of foodborne outbreaks in the United States, and more rapid and efficient detection methods are needed. Surface plasmon resonance imaging (SPRi) is an emerging optical technique, which allows for rapid and label-free screening of multiple targets simultaneous...

  1. Surface plasmon resonance imaging for label-free detection of foodborne pathogens and toxins

    Science.gov (United States)

    More rapid and efficient detection methods for foodborne pathogenic bacteria and toxins are needed to address the long assay time and limitations in multiplex capacity. Surface plasmon resonance imaging (SPRi) is an emerging optical technique, which allows for rapid and label-free screening of multi...

  2. Toward microfluidic sperm refinement: continuous flow label-free analysis and sorting of sperm cells

    NARCIS (Netherlands)

    de Wagenaar, B.; Dekker, Stefan; van den Berg, Albert; Segerink, Loes Irene

    2015-01-01

    This manuscript reports upon the development of a microfluidic setup to detect and sort sperm cells from polystyrene beads label-free and non-invasively. Detection is performed by impedance analysis. When sperm cells passed the microelectrodes, the recorded impedance (19.6 ± 5.7 Ω) was higher

  3. High throughput label-free platform for statistical bio-molecular sensing

    DEFF Research Database (Denmark)

    Bosco, Filippo; Hwu, En-Te; Chen, Ching-Hsiu

    2011-01-01

    Sensors are crucial in many daily operations including security, environmental control, human diagnostics and patient monitoring. Screening and online monitoring require reliable and high-throughput sensing. We report on the demonstration of a high-throughput label-free sensor platform utilizing ...

  4. Label-Free and Real-Time Detection of Protein Ubiquitination with a Biological Nanopore

    NARCIS (Netherlands)

    Wloka, Carsten; Van Meervelt, Veerle; van Gelder, Dewi; Danda, Natasha; Jager, Nienke; Williams, Chris P; Maglia, Giovanni

    The covalent addition of ubiquitin to target proteins is a key post-translational modification that is linked to a myriad of biological processes. Here, we report a fast, single-molecule, and label-free method to probe the ubiquitination of proteins employing an engineered Cytolysin A (ClyA)

  5. Label-free biochips for rapid detection of soybean allergen Glym Bd ...

    African Journals Online (AJOL)

    Label-free biochips for rapid detection of soybean allergen GlymBd 30K (P34) in foods. Wei Wang, Xudong Zhu, Shuang Teng, Qiaojun Fan, Hang Qian. Abstract. Purpose: To develop an innovative method for detection of soybean allergen, Gly mBd 30K (P34) in foods using a biosensor based on high spatial imaging ...

  6. 78 FR 47534 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-08-06

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... directive (AD) 2013-14-51 for General Electric Company (GE) GE90-110B1 and GE90-115B turbofan engines with... all known U.S. owners and operators of GE90-110B1 and GE90-115B turbofan engines. AD 2013-14-51...

  7. Label-free imaging of the native, living cellular nanoarchitecture using partial-wave spectroscopic microscopy

    Science.gov (United States)

    Almassalha, Luay M.; Bauer, Greta M.; Chandler, John E.; Gladstein, Scott; Cherkezyan, Lusik; Stypula-Cyrus, Yolanda; Weinberg, Samuel; Zhang, Di; Thusgaard Ruhoff, Peder; Roy, Hemant K.; Subramanian, Hariharan; Chandel, Navdeep S.; Szleifer, Igal; Backman, Vadim

    2016-01-01

    The organization of chromatin is a regulator of molecular processes including transcription, replication, and DNA repair. The structures within chromatin that regulate these processes span from the nucleosomal (10-nm) to the chromosomal (>200-nm) levels, with little known about the dynamics of chromatin structure between these scales due to a lack of quantitative imaging technique in live cells. Previous work using partial-wave spectroscopic (PWS) microscopy, a quantitative imaging technique with sensitivity to macromolecular organization between 20 and 200 nm, has shown that transformation of chromatin at these length scales is a fundamental event during carcinogenesis. As the dynamics of chromatin likely play a critical regulatory role in cellular function, it is critical to develop live-cell imaging techniques that can probe the real-time temporal behavior of the chromatin nanoarchitecture. Therefore, we developed a live-cell PWS technique that allows high-throughput, label-free study of the causal relationship between nanoscale organization and molecular function in real time. In this work, we use live-cell PWS to study the change in chromatin structure due to DNA damage and expand on the link between metabolic function and the structure of higher-order chromatin. In particular, we studied the temporal changes to chromatin during UV light exposure, show that live-cell DNA-binding dyes induce damage to chromatin within seconds, and demonstrate a direct link between higher-order chromatin structure and mitochondrial membrane potential. Because biological function is tightly paired with structure, live-cell PWS is a powerful tool to study the nanoscale structure–function relationship in live cells. PMID:27702891

  8. A light-sheet microscope compatible with mobile devices for label-free intracellular imaging and biosensing.

    Science.gov (United States)

    Wu, Tsung-Feng; Yen, Tony Minghung; Han, Yuanyuan; Chiu, Yu-Jui; Lin, Eason Y-S; Lo, Yu-Hwa

    2014-09-07

    The inner structure, especially the nuclear structure, of cells carries valuable information about disease and health conditions of a person. Here we demonstrate a label-free technique to enable direct observations and measurements of the size, shape and morphology of the cell nucleus. With a microfabricated lens and a commercial CMOS imager, we form a scanning light-sheet microscope to produce a dark-field optical scattering image of the cell nucleus that overlays with the bright-field image produced in a separate regime of the same CMOS sensor. We have used the device to detect nuclear features that characterize the life cycle of cells and have used the nucleus volume as a new parameter for cell classification. The device can be developed into a portable, low-cost, point-of-care device leveraging the capabilities of the CMOS imagers to be pervasive in mobile electronics.

  9. New Detection Modality for Label-Free Quantification of DNA in Biological Samples via Superparamagnetic Bead Aggregation

    Science.gov (United States)

    Leslie, Daniel C.; Li, Jingyi; Strachan, Briony C.; Begley, Matthew R.; Finkler, David; Bazydlo, Lindsay L.; Barker, N. Scott; Haverstick, Doris; Utz, Marcel; Landers, James P.

    2012-01-01

    Combining DNA and superparamagnetic beads in a rotating magnetic field produces multiparticle aggregates that are visually striking, and enables label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonucleotides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques. PMID:22423674

  10. Direct electroplating of plastic for advanced electrical applications

    DEFF Research Database (Denmark)

    Islam, Aminul; Hansen, Hans Nørgaard; Tang, Peter Torben

    2017-01-01

    for direct electroplating of plastic eliminating the need for slow and expensive processes like electroless metal deposition, PVD coating, painting with conductive inks etc. The results obtained from the test demonstrate the potential of direct electroplating of plastic to enhance the electrical conductivity...

  11. Alternatives to electrical cogeneration: The direct application of steam engines

    International Nuclear Information System (INIS)

    Phillips, W.C.

    1993-01-01

    Although small to medium sized industrial facilities are aware of electrical cogeneration, often they are too small for it to be economically justifiable. The direct application of steam turbine power to equipment formerly powered by electric motors, can allow them to use steam capacity to reduce electrical demand and consumption, bypassing cogeneration. Cogeneration converts the heat energy of steam into circular mechanical motion and then converts the circular mechanical motion into electricity. Each conversion entails a loss of energy due to friction and other conversion losses. A substantial amount of the generated electricity is then converted back into circular motion with electric motors, again incurring energy losses. Directly applying the mechanical motion of turbines eliminates both the motion-to-electricity (generator) and the electricity-to-motion (motor) conversion losses. Excess steam capacity during the summer is not unusual for facilities that use steam to provide winter heating. Similarly, most of these facilities experience a large electrical demand peak during the cooling season due to the electricity needed to operate centrifugal chillers. Steam capacity via a turbine to power the chillers can allow the boilers to operate at a higher loading while reducing electrical consumption and demand precisely those periods when demand reduction is most needed. In facilities where the steam generating capacity is sufficient, air compressors provide an appropriate year-round application for turbine power. This paper is the result of an on-going project by the Energy Division, State of North Carolina, Department of Economic and Community Development, in conjunction with the University of North Carolina at Charlotte. The objective of this project is to educate the operating engineers and managers of small to medium sized manufacturing facilities on the technical application and economic justification of steam turbine power

  12. Determinants of the electric field during transcranial direct current stimulation

    DEFF Research Database (Denmark)

    Opitz, Alexander; Paulus, Walter; Will, Susanne

    2015-01-01

    Transcranial direct current stimulation (tDCS) causes a complex spatial distribution of the electric current flow in the head which hampers the accurate localization of the stimulated brain areas. In this study we show how various anatomical features systematically shape the electric field...... over the motor cortex in small steps to examine the resulting changes of the electric field distribution in the underlying cortex. We examined the effect of skull thickness and composition on the passing currents showing that thinner skull regions lead to higher electric field strengths. This effect...... fluid and the skull, the gyral depth and the distance to the anode and cathode. These factors account for up to 50% of the spatial variation of the electric field strength. Further, we demonstrate that individual anatomical factors can lead to stimulation "hotspots" which are partly resistant...

  13. 77 FR 58471 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2012-09-21

    ... Airworthiness Directives; General Electric Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA... (AD) for General Electric Company GEnx-1B54, GEnx-1B58, GEnx-1B64, GEnx-1B67, GEnx-1B70, GEnx-1B54/P1.../P1, GEnx-1B75/P1, GEnx- 2B67, and GEnx-2B67B turbofan engines. This AD requires initial and...

  14. Label-free identification of individual bacteria using Fourier transform light scattering.

    Science.gov (United States)

    Jo, YoungJu; Jung, JaeHwang; Kim, Min-Hyeok; Park, HyunJoo; Kang, Suk-Jo; Park, YongKeun

    2015-06-15

    Rapid identification of bacterial species is crucial in medicine and food hygiene. In order to achieve rapid and label-free identification of bacterial species at the single bacterium level, we propose and experimentally demonstrate an optical method based on Fourier transform light scattering (FTLS) measurements and statistical classification. For individual rod-shaped bacteria belonging to four bacterial species (Listeria monocytogenes, Escherichia coli, Lactobacillus casei, and Bacillus subtilis), two-dimensional angle-resolved light scattering maps are precisely measured using FTLS technique. The scattering maps are then systematically analyzed, employing statistical classification in order to extract the unique fingerprint patterns for each species, so that a new unidentified bacterium can be identified by a single light scattering measurement. The single-bacterial and label-free nature of our method suggests wide applicability for rapid point-of-care bacterial diagnosis.

  15. Artificial intelligence in label-free microscopy biological cell classification by time stretch

    CERN Document Server

    Mahjoubfar, Ata; Jalali, Bahram

    2017-01-01

    This book introduces time-stretch quantitative phase imaging (TS-QPI), a high-throughput label-free imaging flow cytometer developed for big data acquisition and analysis in phenotypic screening. TS-QPI is able to capture quantitative optical phase and intensity images simultaneously, enabling high-content cell analysis, cancer diagnostics, personalized genomics, and drug development. The authors also demonstrate a complete machine learning pipeline that performs optical phase measurement, image processing, feature extraction, and classification, enabling high-throughput quantitative imaging that achieves record high accuracy in label -free cellular phenotypic screening and opens up a new path to data-driven diagnosis. • Demonstrates how machine learning is used in high-speed microscopy imaging to facilitate medical diagnosis; • Provides a systematic and comprehensive illustration of time stretch technology; • Enables multidisciplinary application, including industrial, biomedical, and artificial intell...

  16. Improving Probe Immobilization for Label-Free Capacitive Detection of DNA Hybridization on Microfabricated Gold Electrodes

    Directory of Open Access Journals (Sweden)

    Sandro Carrara

    2008-02-01

    Full Text Available Alternative approaches to labeled optical detection for DNA arrays are actively investigated for low-cost point-of-care applications. In this domain, label-free capacitive detection is one of the most intensely studied techniques. It is based on the idea to detect the Helmholtz ion layer displacements when molecular recognition occurs at the electrodes/solution interface. The sensing layer is usually prepared by using thiols terminated DNA single-strength oligonucleotide probes on top of the sensor electrodes. However, published data shows evident time drift, which greatly complicates signal conditioning and processing and ultimately increases the uncertainty in DNA recognition sensing. The aim of this work is to show that newly developed ethylene-glycol functionalized alkanethiols greatly reduce time drift, thereby significantly improving capacitance based label-free detection of DNA.

  17. Label free imaging of cell-substrate contacts by holographic total internal reflection microscopy.

    Science.gov (United States)

    Mandracchia, Biagio; Gennari, Oriella; Marchesano, Valentina; Paturzo, Melania; Ferraro, Pietro

    2017-09-01

    The study of cell adhesion contacts is pivotal to understand cell mechanics and interaction at substrates or chemical and physical stimuli. We designed and built a HoloTIR microscope for label-free quantitative phase imaging of total internal reflection. Here we show for the first time that HoloTIR is a good choice for label-free study of focal contacts and of cell/substrate interaction as its sensitivity is enhanced in comparison with standard TIR microscopy. Finally, the simplicity of implementation and relative low cost, due to the requirement of less optical components, make HoloTIR a reasonable alternative, or even an addition, to TIRF microscopy for mapping cell/substratum topography. As a proof of concept, we studied the formation of focal contacts of fibroblasts on three substrates with different levels of affinity for cell adhesion. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Label-free proteomic methodology for the analysis of human kidney stone matrix composition

    DEFF Research Database (Denmark)

    Witzmann, Frank A; Evan, Andrew P; Coe, Fredric L

    2016-01-01

    to generate a comprehensive profile of their proteomes, including physicochemical and bioinformatic analysis.`. RESULTS: We identified and quantified 1,059 unique protein database entries in the two human kidney stone samples, revealing a more complex proteome than previously reported. Protein composition...... demonstrate relevant constitutive differences between stones. We have analyzed the matrices of two unique human calcium oxalate stones (CaOx-Ia and CaOx-Id) using a simple but effective chaotropic reducing solution for extraction/solubilization combined with label-free quantitative mass spectrometry...... powders combined with label-free quantitative mass spectrometry has yielded the most comprehensive list to date of the proteins that constitute the human kidney stone proteome....

  19. Optimization of High-Q Coupled Nanobeam Cavity for Label-Free Sensing

    OpenAIRE

    Yaseen, Mohammad; Yang, Yi-Chun; Shih, Min-Hsiung; Chang, Yia-Chung

    2015-01-01

    We numerically and experimentally investigated the lateral coupling between photonic crystal (PhC) nanobeam (NB) cavities, pursuing high sensitivity and figure of merit (FOM) label-free biosensor. We numerically carried out 3D finite-difference time-domain (3D-FDTD) and the finite element method (FEM) simulations. We showed that when two PhC NB cavities separated by a small gap are evanescently coupled, the variation in the gap width significantly changes the coupling efficiency between the ...

  20. A Fast Colourimetric Assay for Lead Detection Using Label-Free Gold Nanoparticles (AuNPs

    Directory of Open Access Journals (Sweden)

    Guowei Zhong

    2015-04-01

    Full Text Available A sensitive colourimetric method for lead (PbII detection is reported in this paper using a common tripeptide, glutathione (GSH, and label-free gold nanoparticles (AuNPs. A limit of detection of 6.0 ppb in water was achieved and the dynamic linear range was up to 500 ppb. Selectivity over fourteen potential interfering metal ions was tested and most of these metal ions do not interfere with the method.

  1. Label-free histamine detection with nanofluidic diodes through metal ion displacement mechanism

    OpenAIRE

    Ali, Mubarak; Ramirez Hoyos, Patricio; Duznovic, Ivana; Nasir, Saima; Mafe, Salvador; Ensinger, Wolfgang

    2017-01-01

    [EN] We design and characterize a nanofluidic device for the label-free specific detection of histamine neurotransmitter based on a metal ion displacement mechanism. The sensor consists of an asymmetric polymer nanopore fabricated via ion track-etching technique. The nanopore sensor surface having metal-nitrilotriacetic (NTA-Ni2+) chelates is obtained by covalent coupling of native carboxylic acid groups with N-alpha,N-alpha-bis(carboxymethyl)-L-lysine (BCML), followed by exposure to Ni2+ ion...

  2. Accurate label-free reaction kinetics determination using initial rate heat measurements

    Science.gov (United States)

    Ebrahimi, Kourosh Honarmand; Hagedoorn, Peter-Leon; Jacobs, Denise; Hagen, Wilfred R.

    2015-01-01

    Accurate label-free methods or assays to obtain the initial reaction rates have significant importance in fundamental studies of enzymes and in application-oriented high throughput screening of enzyme activity. Here we introduce a label-free approach for obtaining initial rates of enzyme activity from heat measurements, which we name initial rate calorimetry (IrCal). This approach is based on our new finding that the data recorded by isothermal titration calorimetry for the early stages of a reaction, which have been widely ignored, are correlated to the initial rates. Application of the IrCal approach to various enzymes led to accurate enzyme kinetics parameters as compared to spectroscopic methods and enabled enzyme kinetic studies with natural substrate, e.g. proteases with protein substrates. Because heat is a label-free property of almost all reactions, the IrCal approach holds promise in fundamental studies of various enzymes and in use of calorimetry for high throughput screening of enzyme activity. PMID:26574737

  3. Multimodal nonlinear microscopy: A powerful label-free method for supporting standard diagnostics on biological tissues

    Directory of Open Access Journals (Sweden)

    Riccardo Cicchi

    2014-09-01

    Full Text Available The large use of nonlinear laser scanning microscopy in the past decade paved the way for potential clinical application of this imaging technique. Modern nonlinear microscopy techniques offer promising label-free solutions to improve diagnostic performances on tissues. In particular, the combination of multiple nonlinear imaging techniques in the same microscope allows integrating morphological with functional information in a morpho-functional scheme. Such approach provides a high-resolution label-free alternative to both histological and immunohistochemical examination of tissues and is becoming increasingly popular among the clinical community. Nevertheless, several technical improvements, including automatic scanning and image analysis, are required before the technique represents a standard diagnostic method. In this review paper, we highlight the capabilities of multimodal nonlinear microscopy for tissue imaging, by providing various examples on colon, arterial and skin tissues. The comparison between images acquired using multimodal nonlinear microscopy and histology shows a good agreement between the two methods. The results demonstrate that multimodal nonlinear microscopy is a powerful label-free alternative to standard histopathological methods and has the potential to find a stable place in the clinical setting in the near future.

  4. freeQuant: A Mass Spectrometry Label-Free Quantification Software Tool for Complex Proteome Analysis.

    Science.gov (United States)

    Deng, Ning; Li, Zhenye; Pan, Chao; Duan, Huilong

    2015-01-01

    Study of complex proteome brings forward higher request for the quantification method using mass spectrometry technology. In this paper, we present a mass spectrometry label-free quantification tool for complex proteomes, called freeQuant, which integrated quantification with functional analysis effectively. freeQuant consists of two well-integrated modules: label-free quantification and functional analysis with biomedical knowledge. freeQuant supports label-free quantitative analysis which makes full use of tandem mass spectrometry (MS/MS) spectral count, protein sequence length, shared peptides, and ion intensity. It adopts spectral count for quantitative analysis and builds a new method for shared peptides to accurately evaluate abundance of isoforms. For proteins with low abundance, MS/MS total ion count coupled with spectral count is included to ensure accurate protein quantification. Furthermore, freeQuant supports the large-scale functional annotations for complex proteomes. Mitochondrial proteomes from the mouse heart, the mouse liver, and the human heart were used to evaluate the usability and performance of freeQuant. The evaluation showed that the quantitative algorithms implemented in freeQuant can improve accuracy of quantification with better dynamic range.

  5. Label-free quantitative proteomics of CD133-positive liver cancer stem cells

    Directory of Open Access Journals (Sweden)

    Tsai Sheng-Ta

    2012-11-01

    Full Text Available Abstract Background CD133-positive liver cancer stem cells, which are characterized by their resistance to conventional chemotherapy and their tumor initiation ability at limited dilutions, have been recognized as a critical target in liver cancer therapeutics. In the current work, we developed a label-free quantitative method to investigate the proteome of CD133-positive liver cancer stem cells for the purpose of identifying unique biomarkers that can be utilized for targeting liver cancer stem cells. Label-free quantitation was performed in combination with ID-based Elution time Alignment by Linear regression Quantitation (IDEAL-Q and MaxQuant. Results Initially, IDEAL-Q analysis revealed that 151 proteins were differentially expressed in the CD133-positive hepatoma cells when compared with CD133-negative cells. We then analyzed these 151 differentially expressed proteins by MaxQuant software and identified 10 significantly up-regulated proteins. The results were further validated by RT-PCR, western blot, flow cytometry or immunofluorescent staining which revealed that prominin-1, annexin A1, annexin A3, transgelin, creatine kinase B, vimentin, and EpCAM were indeed highly expressed in the CD133-positive hepatoma cells. Conclusions These findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.

  6. Reproducible E. coli detection based on label-free SERS and mapping.

    Science.gov (United States)

    Yang, Danting; Zhou, Haibo; Haisch, Christoph; Niessner, Reinhard; Ying, Yibin

    2016-01-01

    The biosensing for rapid detection of bacteria based on surface-enhanced Raman scattering (SERS) has been widely explored for recent years. It is still a challenge to achieve a high sensitive, reproducible label free detection method for bacteria. In this work, a label-free SERS detection method of Escherichia coli based on incubation with silver colloid was reported. Optimized incubation conditions including shaking speed, time and temperature were used to help construct a rapid SERS method for E. coli analysis. It was found that the enhancement of the Raman signal of E. coli could be achieved to 1.8×10(4) cps (counts per second) with high reproducibility. Three strains of E. coli DSM 1116/498/5695 could be successfully discriminated using such SERS method combining discriminant analysis. Finally, the lowest concentration of E. coli at 1×10(5) cell/mL can be detected by SERS mapping. Thus, our detection method offers higher sensitivity and reproducibility compared to previously reported label free simple-mixing methods, opening an avenue for developing various SERS-based biosensor. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Label-free capture of breast cancer cells spiked in buffy coats using carbon nanotube antibody micro-arrays

    Science.gov (United States)

    Khosravi, Farhad; Trainor, Patrick; Rai, Shesh N.; Kloecker, Goetz; Wickstrom, Eric; Panchapakesan, Balaji

    2016-04-01

    We demonstrate the rapid and label-free capture of breast cancer cells spiked in buffy coats using nanotube-antibody micro-arrays. Single wall carbon nanotube arrays were manufactured using photo-lithography, metal deposition, and etching techniques. Anti-epithelial cell adhesion molecule (EpCAM) antibodies were functionalized to the surface of the nanotube devices using 1-pyrene-butanoic acid succinimidyl ester functionalization method. Following functionalization, plain buffy coat and MCF7 cell spiked buffy coats were adsorbed on to the nanotube device and electrical signatures were recorded for differences in interaction between samples. A statistical classifier for the ‘liquid biopsy’ was developed to create a predictive model based on dynamic time warping to classify device electrical signals that corresponded to plain (control) or spiked buffy coats (case). In training test, the device electrical signals originating from buffy versus spiked buffy samples were classified with ˜100% sensitivity, ˜91% specificity and ˜96% accuracy. In the blinded test, the signals were classified with ˜91% sensitivity, ˜82% specificity and ˜86% accuracy. A heatmap was generated to visually capture the relationship between electrical signatures and the sample condition. Confocal microscopic analysis of devices that were classified as spiked buffy coats based on their electrical signatures confirmed the presence of cancer cells, their attachment to the device and overexpression of EpCAM receptors. The cell numbers were counted to be ˜1-17 cells per 5 μl per device suggesting single cell sensitivity in spiked buffy coats that is scalable to higher volumes using the micro-arrays.

  8. Electric vehicle drive train with direct coupling transmission

    Science.gov (United States)

    Tankersley, Jerome B.; Boothe, Richard W.; Konrad, Charles E.

    1995-01-01

    An electric vehicle drive train includes an electric motor and an associated speed sensor, a transmission operable in a speed reduction mode or a direct coupled mode, and a controller responsive to the speed sensor for operating the transmission in the speed reduction mode when the motor is below a predetermined value, and for operating the motor in the direct coupled mode when the motor speed is above a predetermined value. The controller reduces the speed of the motor, such as by regeneratively braking the motor, when changing from the speed reduction mode to the direct coupled mode. The motor speed may be increased when changing from the direct coupled mode to the speed reduction mode. The transmission is preferably a single stage planetary gearbox.

  9. Electric field-induced astrocyte alignment directs neurite outgrowth.

    Science.gov (United States)

    Alexander, John K; Fuss, Babette; Colello, Raymond J

    2006-05-01

    The extension and directionality of neurite outgrowth are key to achieving successful target connections during both CNS development and during the re-establishment of connections lost after neural trauma. The degree of axonal elongation depends, in large part, on the spatial arrangement of astrocytic processes rich in growth-promoting proteins. Because astrocytes in culture align their processes on exposure to an electrical field of physiological strength, we sought to determine the extent to which aligned astrocytes affect neurite outgrowth. To this end, dorsal root ganglia cells were seeded onto cultured rat astrocytes that were pre-aligned by exposure to an electric field of physiological strength (500 mV mm(-1)). Using confocal microscopy and digital image analysis, we found that neurite outgrowth at 24 hours and at 48 hours is enhanced significantly and directed consistently along the aligned astrocyte processes. Moreover, this directed neurite outgrowth is maintained when grown on fixed, aligned astrocytes. Collectively, these results indicate that endogenous electric fields present within the developing CNS might act to align astrocyte processes, which can promote and direct neurite growth. Furthermore, these results demonstrate a simple method to produce an aligned cellular substrate, which might be used to direct regenerating neurites.

  10. Design optimization of structural parameters for highly sensitive photonic crystal label-free biosensors.

    Science.gov (United States)

    Ju, Jonghyun; Han, Yun-ah; Kim, Seok-min

    2013-03-07

    The effects of structural design parameters on the performance of nano-replicated photonic crystal (PC) label-free biosensors were examined by the analysis of simulated reflection spectra of PC structures. The grating pitch, duty, scaled grating height and scaled TiO2 layer thickness were selected as the design factors to optimize the PC structure. The peak wavelength value (PWV), full width at half maximum of the peak, figure of merit for the bulk and surface sensitivities, and surface/bulk sensitivity ratio were also selected as the responses to optimize the PC label-free biosensor performance. A parametric study showed that the grating pitch was the dominant factor for PWV, and that it had low interaction effects with other scaled design factors. Therefore, we can isolate the effect of grating pitch using scaled design factors. For the design of PC-label free biosensor, one should consider that: (1) the PWV can be measured by the reflection peak measurement instruments, (2) the grating pitch and duty can be manufactured using conventional lithography systems, and (3) the optimum design is less sensitive to the grating height and TiO2 layer thickness variations in the fabrication process. In this paper, we suggested a design guide for highly sensitive PC biosensor in which one select the grating pitch and duty based on the limitations of the lithography and measurement system, and conduct a multi objective optimization of the grating height and TiO2 layer thickness for maximizing performance and minimizing the influence of parameter variation. Through multi-objective optimization of a PC structure with a fixed grating height of 550 nm and a duty of 50%, we obtained a surface FOM of 66.18 RIU-1 and an S/B ratio of 34.8%, with a grating height of 117 nm and TiO2 height of 210 nm.

  11. Design Optimization of Structural Parameters for Highly Sensitive Photonic Crystal Label-Free Biosensors

    Directory of Open Access Journals (Sweden)

    Yun-ah Han

    2013-03-01

    Full Text Available The effects of structural design parameters on the performance of nano-replicated photonic crystal (PC label-free biosensors were examined by the analysis of simulated reflection spectra of PC structures. The grating pitch, duty, scaled grating height and scaled TiO2 layer thickness were selected as the design factors to optimize the PC structure. The peak wavelength value (PWV, full width at half maximum of the peak, figure of merit for the bulk and surface sensitivities, and surface/bulk sensitivity ratio were also selected as the responses to optimize the PC label-free biosensor performance. A parametric study showed that the grating pitch was the dominant factor for PWV, and that it had low interaction effects with other scaled design factors. Therefore, we can isolate the effect of grating pitch using scaled design factors. For the design of PC-label free biosensor, one should consider that: (1 the PWV can be measured by the reflection peak measurement instruments, (2 the grating pitch and duty can be manufactured using conventional lithography systems, and (3 the optimum design is less sensitive to the grating height and TiO2 layer thickness variations in the fabrication process. In this paper, we suggested a design guide for highly sensitive PC biosensor in which one select the grating pitch and duty based on the limitations of the lithography and measurement system, and conduct a multi objective optimization of the grating height and TiO2 layer thickness for maximizing performance and minimizing the influence of parameter variation. Through multi-objective optimization of a PC structure with a fixed grating height of 550 nm and a duty of 50%, we obtained a surface FOM of 66.18 RIU−1 and an S/B ratio of 34.8%, with a grating height of 117 nm and TiO2 height of 210 nm.

  12. Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing

    OpenAIRE

    J. Riba; T. Gleichmann; S. Zimmermann; R. Zengerle; P. Koltay

    2016-01-01

    The isolation and analysis of single prokaryotic cells down to 1??m and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35?pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20??m in size....

  13. Comparison of serum fractionation methods by data independent label-free proteomics

    Directory of Open Access Journals (Sweden)

    D. Baiwir

    2015-12-01

    Full Text Available Off-line sample prefractionations applied prior to biomarker discovery proteomics are options to enable more protein identifications and detect low-abundance proteins. This work compared five commercial methods efficiency to raw serum analysis using label-free proteomics. The variability of the protein quantities determined for each process was similar to the unprefractionated serum. A 49% increase in protein identifications and 12.2% of reliable quantification were obtained. A 61 times lower limit of protein quantitation was reached compared to protein concentrations observed in raw serum. The concentrations of detected proteins were confronted to estimated reference values.

  14. Quantitative Label-Free Cell Proliferation Tracking with a Versatile Electrochemical Impedance Detection Platform

    DEFF Research Database (Denmark)

    Caviglia, Claudia; Carminati, M; Heiskanen, Arto

    2012-01-01

    Since the use of impedance measurements for label-free monitoring of cells has become widespread but still the choice of sensing configuration is not unique though crucial for a quantitative interpretation of data, we demonstrate the application of a novel custom multipotentiostat platform to study...... optimal detection strategies. Electrochemical Impedance Spectroscopy (EIS) has been used to monitor and compare adhesion of different cell lines. HeLa cells and 3T3 fibroblasts have been cultured for 12 hours on interdigitated electrode arrays integrated into a tailor-made cell culture platform. Both...... vertical and coplanar interdigitated sensing configuration approaches have been used and compared on the same cell populations....

  15. Label-free three-dimensional imaging of cell nucleus using third-harmonic generation microscopy

    Science.gov (United States)

    Lin, Jian; Zheng, Wei; Wang, Zi; Huang, Zhiwei

    2014-09-01

    We report the implementation of the combined third-harmonic generation (THG) and two-photon excited fluorescence (TPEF) microscopy for label-free three-dimensional (3-D) imaging of cell nucleus morphological changes in liver tissue. THG imaging shows regular spherical shapes of normal hepatocytes nuclei with inner chromatin structures while revealing the condensation of chromatins and nuclear fragmentations in hepatocytes of diseased liver tissue. Colocalized THG and TPEF imaging provides complementary information of cell nuclei and cytoplasm in tissue. This work suggests that 3-D THG microscopy has the potential for quantitative analysis of nuclear morphology in cells at a submicron-resolution without the need for DNA staining.

  16. Label-free three-dimensional imaging of cell nucleus using third-harmonic generation microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Jian; Zheng, Wei; Wang, Zi; Huang, Zhiwei, E-mail: biehzw@nus.edu.sg [Optical Bioimaging Laboratory, Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore, Singapore 117576 (Singapore)

    2014-09-08

    We report the implementation of the combined third-harmonic generation (THG) and two-photon excited fluorescence (TPEF) microscopy for label-free three-dimensional (3-D) imaging of cell nucleus morphological changes in liver tissue. THG imaging shows regular spherical shapes of normal hepatocytes nuclei with inner chromatin structures while revealing the condensation of chromatins and nuclear fragmentations in hepatocytes of diseased liver tissue. Colocalized THG and TPEF imaging provides complementary information of cell nuclei and cytoplasm in tissue. This work suggests that 3-D THG microscopy has the potential for quantitative analysis of nuclear morphology in cells at a submicron-resolution without the need for DNA staining.

  17. A label free aptamer-based LPG sensor for detection of mercury in aquatic solutions

    Science.gov (United States)

    Nikbakht, Hamed; Latifi, Hamid; Ziaee, Farzaneh

    2015-09-01

    We demonstrate a label free fiber optic sensor for detection of mercury ions in aquatic solutions. This sensor utilizes aptamers as bio-recognition element which traps mercury ions and cause a refractive index change in the vicinity of the sensor. Refractive index variations lead to a change in the transmission spectrum that can be used to calculate the concentration of mercury ions in that solution. The concentration of 1 nM mercury ions was detected which is below the specific amount determined by the US environmental protection agency as the maximum authorized contaminant level of Hg2+ ions in drinking water.

  18. A comparative study of fat storage quantitation in nematode Caenorhabditis elegans using label and label-free methods.

    Directory of Open Access Journals (Sweden)

    Kelvin Yen

    Full Text Available The nematode Caenorhabditis elegans has been employed as a model organism to study human obesity due to the conservation of the pathways that regulate energy metabolism. To assay for fat storage in C. elegans, a number of fat-soluble dyes have been employed including BODIPY, Nile Red, Oil Red O, and Sudan Black. However, dye-labeled assays produce results that often do not correlate with fat stores in C. elegans. An alternative label-free approach to analyze fat storage in C. elegans has recently been described with coherent anti-Stokes Raman scattering (CARS microscopy. Here, we compare the performance of CARS microscopy with standard dye-labeled techniques and biochemical quantification to analyze fat storage in wild type C. elegans and with genetic mutations in the insulin/IGF-1 signaling pathway including the genes daf-2 (insulin/IGF-1 receptor, rict-1 (rictor and sgk-1 (serum glucocorticoid kinase. CARS imaging provides a direct measure of fat storage with unprecedented details including total fat stores as well as the size, number, and lipid-chain unsaturation of individual lipid droplets. In addition, CARS/TPEF imaging reveals a neutral lipid species that resides in both the hypodermis and the intestinal cells and an autofluorescent organelle that resides exclusively in the intestinal cells. Importantly, coherent addition of the CARS fields from the C-H abundant neutral lipid permits selective CARS imaging of the fat store, and further coupling of spontaneous Raman analysis provides unprecedented details including lipid-chain unsaturation of individual lipid droplets. We observe that although daf-2, rict-1, and sgk-1 mutants affect insulin/IGF-1 signaling, they exhibit vastly different phenotypes in terms of neutral lipid and autofluorescent species. We find that CARS imaging gives quantification similar to standard biochemical triglyceride quantification. Further, we independently confirm that feeding worms with vital dyes does not lead

  19. A Comparative Study of Fat Storage Quantitation in Nematode Caenorhabditis elegans Using Label and Label-Free Methods

    Science.gov (United States)

    Bansal, Ankita; Narasimhan, Sri Devi; Cheng, Ji-Xin; Tissenbaum, Heidi A.

    2010-01-01

    The nematode Caenorhabditis elegans has been employed as a model organism to study human obesity due to the conservation of the pathways that regulate energy metabolism. To assay for fat storage in C. elegans, a number of fat-soluble dyes have been employed including BODIPY, Nile Red, Oil Red O, and Sudan Black. However, dye-labeled assays produce results that often do not correlate with fat stores in C. elegans. An alternative label-free approach to analyze fat storage in C. elegans has recently been described with coherent anti-Stokes Raman scattering (CARS) microscopy. Here, we compare the performance of CARS microscopy with standard dye-labeled techniques and biochemical quantification to analyze fat storage in wild type C. elegans and with genetic mutations in the insulin/IGF-1 signaling pathway including the genes daf-2 (insulin/IGF-1 receptor), rict-1 (rictor) and sgk-1 (serum glucocorticoid kinase). CARS imaging provides a direct measure of fat storage with unprecedented details including total fat stores as well as the size, number, and lipid-chain unsaturation of individual lipid droplets. In addition, CARS/TPEF imaging reveals a neutral lipid species that resides in both the hypodermis and the intestinal cells and an autofluorescent organelle that resides exclusively in the intestinal cells. Importantly, coherent addition of the CARS fields from the C-H abundant neutral lipid permits selective CARS imaging of the fat store, and further coupling of spontaneous Raman analysis provides unprecedented details including lipid-chain unsaturation of individual lipid droplets. We observe that although daf-2, rict-1, and sgk-1 mutants affect insulin/IGF-1 signaling, they exhibit vastly different phenotypes in terms of neutral lipid and autofluorescent species. We find that CARS imaging gives quantification similar to standard biochemical triglyceride quantification. Further, we independently confirm that feeding worms with vital dyes does not lead to the staining

  20. Electric field-induced astrocyte alignment directs neurite outgrowth

    OpenAIRE

    ALEXANDER, JOHN K.; FUSS, BABETTE; COLELLO, RAYMOND J.

    2006-01-01

    The extension and directionality of neurite outgrowth are key to achieving successful target connections during both CNS development and during the re-establishment of connections lost after neural trauma. The degree of axonal elongation depends, in large part, on the spatial arrangement of astrocytic processes rich in growth-promoting proteins. Because astrocytes in culture align their processes on exposure to an electrical field of physiological strength, we sought to determine the extent t...

  1. Label-free DNA-based detection of Mycobacterium tuberculosis and rifampicin resistance through hydration induced stress in microcantilevers.

    Science.gov (United States)

    Domínguez, Carmen M; Kosaka, Priscila M; Sotillo, Alma; Mingorance, Jesús; Tamayo, Javier; Calleja, Montserrat

    2015-02-03

    We have developed a label-free assay for the genomic detection of Mycobacterium tuberculosis and rifampicin resistance. The method relies on the quantification of the hydration induced stress on microcantilever biosensors functionalized with oligonucleotide probes, before and after hybridization with specific targets. We have found a limit of detection of 10 fg/mL for PCR amplified products of 122 bp. Furthermore, the technique can successfully target genomic DNA (gDNA) fragments of length >500 bp, and it can successfully discriminate single mismatches. We have used both loci IS6110 and rpoB as targets to detect the mycobacteria and the rifampicin resistance from gDNA directly extracted from bacterial culture and without PCR amplification. We have been able to detect 2 pg/mL target concentration in samples with an excess of interfering DNA and in a total analysis time of 1 h and 30 min. The detection limit found demonstrates the capability to develop direct assays without the need for long culture steps or PCR amplification. The methodology can be easily translated to different microbial targets, and it is suitable for further development of miniaturized devices and multiplexed detection.

  2. Label-free detection of aflatoxin M1 with electrochemical Fe{sub 3}O{sub 4}/polyaniline-based aptasensor

    Energy Technology Data Exchange (ETDEWEB)

    Nguyen, Binh Hai [Institute of Material Science, Vietnam Academy of Science and Technology, 18, Hoang Quoc Viet Road, Cau Giay, Hanoi (Viet Nam); Tran, Lam Dai, E-mail: lamtd@ims.vast.ac.vn [Institute of Material Science, Vietnam Academy of Science and Technology, 18, Hoang Quoc Viet Road, Cau Giay, Hanoi (Viet Nam); Do, Quan Phuc [Research Center for Environmental Technology and Sustainable Development, Hanoi University of Science, Hanoi (Viet Nam); Nguyen, Huy Le [School of Chemical Engineering, Hanoi University of Science and Technology, 1, Dai Co Viet Road, Hanoi (Viet Nam); Tran, Ngoc Huan [Department of Chemistry, Hanyang University, Seoul 133-791 (Korea, Republic of); Nguyen, Phuc Xuan [Institute of Material Science, Vietnam Academy of Science and Technology, 18, Hoang Quoc Viet Road, Cau Giay, Hanoi (Viet Nam)

    2013-05-01

    The selective detection of ultratrace amounts of aflatoxin M1 (AFM1) is extremely important for food safety since it is the most toxic mycotoxin class that is allowed to be present on cow milk with strictly low regulatory levels. In this work, Fe{sub 3}O{sub 4} incorporated polyaniline (Fe{sub 3}O{sub 4}/PANi) film has been polymerized on interdigitated electrode (IDE) as sensitive film for AFM1 electrochemical biosensor. The immobilized aptamers as an affinity capture reagent and magnetic nanoparticles for signal amplification element have been employed in the sensing platform. Label-free and direct detection of the aptamer-AFM1 on Fe{sub 3}O{sub 4}/PANi interface were performed via electrochemical signal change, acquired by cyclic and square wave voltammetries. With a simplified strategy, this electrochemical aptasensor shows a good sensitivity to AFM1 in the range of 6–60 ng·L{sup −1}, with the detection limit of 1.98 ng·L{sup −1}. The results open up the path for designing cost effective aptasensors for other biomedical applications. Highlights: ► Label-free detection of the aptamer-AFM1 on Fe{sub 3}O{sub 4}/PANi interface ► Electrochemical aptasensor with a good sensitivity to AFM1 in the range of 6–60 ng·L{sup −1}, with detection limit of 1.98 ng·L{sup −1} ► Advantages over other analytical techniques in terms of label free format, sensitivity, stability and analysis time.

  3. Label-free determination of hemodynamic parameters in the microcirculaton with third harmonic generation microscopy.

    Directory of Open Access Journals (Sweden)

    Steffen Dietzel

    Full Text Available Determination of blood flow velocity and related hemodynamic parameters is an important aspect of physiological studies which in many settings requires fluorescent labeling. Here we show that Third Harmonic Generation (THG microscopy is a suitable tool for label-free intravital investigations of the microcirculation in widely-used physiological model systems. THG microscopy is a non-fluorescent multi-photon scanning technique combining the advantages of label-free imaging with restriction of signal generation to a focal spot. Blood flow was visualized and its velocity was measured in adult mouse cremaster muscle vessels, non-invasively in mouse ear vessels and in Xenopus tadpoles. In arterioles, THG line scanning allowed determination of the flow pulse velocity curve and hence the heart rate. By relocating the scan line we obtained velocity profiles through vessel diameters, allowing shear rate calculations. The cell free layer containing the glycocalyx was also visualized. Comparison of the current microscopic resolution with theoretical, diffraction limited resolution let us conclude that an about sixty-fold THG signal intensity increase may be possible with future improved optics, optimized for 1200-1300 nm excitation. THG microscopy is compatible with simultaneous two-photon excited fluorescence detection. It thus also provides the opportunity to determine important hemodynamic parameters in parallel to common fluorescent observations without additional label.

  4. A single-layer, planar, optofluidic Mach–Zehnder interferometer for label-free detection†

    Science.gov (United States)

    Lapsley, Michael Ian; Chiang, I.-Kao; Zheng, Yue Bing; Ding, Xiaoyun; Mao, Xiaole

    2014-01-01

    We have developed a planar, optofluidic Mach–Zehnder interferometer for the label-free detection of liquid samples. In contrast to most on-chip interferometers which require complex fabrication, our design was realized via a simple, single-layer soft lithography fabrication process. In addition, a single-wavelength laser source and a silicon photodetector were the only optical equipment used for data collection. The device was calibrated using published data for the refractive index of calcium chloride (CaCl2) in solution, and the biosensing capabilities of the device were tested by detecting bovine serum albumin (BSA). Our design enables a refractometer with a low limit of detection (1.24 × 10−4 refractive index units (RIU)), low variability (1 × 10−4 RIU), and high sensitivity (927.88 oscillations per RIU). This performance is comparable to state-of-the-art optofluidic refractometers that involve complex fabrication processes and/or expensive, bulky optics. The advantages of our device (i.e. simple fabrication process, straightforward optical equipment, low cost, and high detection sensitivity) make it a promising candidate for future mass-producible, inexpensive, highly sensitive, label-free optical detection systems. PMID:21479332

  5. A single-layer, planar, optofluidic Mach-Zehnder interferometer for label-free detection.

    Science.gov (United States)

    Lapsley, Michael Ian; Chiang, I-Kao; Zheng, Yue Bing; Ding, Xiaoyun; Mao, Xiaole; Huang, Tony Jun

    2011-05-21

    We have developed a planar, optofluidic Mach-Zehnder interferometer for the label-free detection of liquid samples. In contrast to most on-chip interferometers which require complex fabrication, our design was realized via a simple, single-layer soft lithography fabrication process. In addition, a single-wavelength laser source and a silicon photodetector were the only optical equipment used for data collection. The device was calibrated using published data for the refractive index of calcium chloride (CaCl(2)) in solution, and the biosensing capabilities of the device were tested by detecting bovine serum albumin (BSA). Our design enables a refractometer with a low limit of detection (1.24 × 10(-4) refractive index units (RIU)), low variability (1 × 10(-4) RIU), and high sensitivity (927.88 oscillations per RIU). This performance is comparable to state-of-the-art optofluidic refractometers that involve complex fabrication processes and/or expensive, bulky optics. The advantages of our device (i.e. simple fabrication process, straightforward optical equipment, low cost, and high detection sensitivity) make it a promising candidate for future mass-producible, inexpensive, highly sensitive, label-free optical detection systems. © The Royal Society of Chemistry 2011

  6. A combinatorial approach to the peptide feature matching problem for label-free quantification.

    Science.gov (United States)

    Lin, Hao; He, Lin; Ma, Bin

    2013-07-15

    Label-free quantification is an important approach to identify biomarkers, as it measures the quantity change of peptides across different biological samples. One of the fundamental steps for label-free quantification is to match the peptide features that are detected in two datasets to each other. Although ad hoc software tools exist for the feature matching, the definition of a combinatorial model for this problem is still not available. A combinatorial model is proposed in this article. Each peptide feature contains a mass value and a retention time value, which are used to calculate a matching weight between a pair of features. The feature matching is to find the maximum-weighted matching between the two sets of features, after applying a to-be-computed time alignment function to all the retention time values of one set of the features. This is similar to the maximum matching problem in a bipartite graph. But we show that the requirement of time alignment makes the problem NP-hard. Practical algorithms are also provided. Experiments on real data show that the algorithm compares favorably with other existing methods. binma@uwaterloo.ca Supplementary data are available at Bioinformatics online.

  7. A label-free electrochemiluminescent sensor for ATP detection based on ATP-dependent ligation.

    Science.gov (United States)

    Zhao, Tingting; Lin, Chunshui; Yao, Qiuhong; Chen, Xi

    2016-07-01

    In this work, we describe a new label-free, sensitive and highly selective strategy for the electrochemiluminescent (ECL) detection of ATP at the picomolar level via ATP-induced ligation. The molecular-beacon like DNA probes (P12 complex) are self-assembled on a gold electrode. The presence of ATP leads to the ligation of P12 complex which blocks the digestion by Exonuclease III (Exo III). The protected P12 complex causes the intercalation of numerous ECL indicators (Ru(phen)3(2+)) into the duplex DNA grooves, resulting in significantly amplified ECL signal output. Since the ligating site of T4 DNA ligase and the nicking site of Exo III are the same, it involves no long time of incubation for conformation change. The proposed strategy combines the amplification power of enzyme and the inherent high sensitivity of the ECL technique and enables picomolar detection of ATP. The developed strategy also shows high selectivity against ATP analogs, which makes our new label-free and highly sensitive ligation-based method a useful addition to the amplified ATP detection arena. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Label-free amino acid detection based on nanocomposites of graphene oxide hybridized with gold nanoparticles.

    Science.gov (United States)

    Zhang, Qian; Zhang, Diming; Lu, Yanli; Xu, Gang; Yao, Yao; Li, Shuang; Liu, Qingjun

    2016-03-15

    Nanocomposites of graphene oxide and gold nanoparticles (GO/GNPs) were synthesized for label-free detections of amino acids. Interactions between the composites and amino acids were investigated by both naked-eye observation and optical absorption spectroscopy. The GO/GNPs composites displayed apparent color changes and absorption spectra changes in presences of amino acids including glutamate, aspartate, and cysteine. The interaction mechanisms of the composites and amino acids were discussed and explored with sulfhydryl groups and non-α-carboxylic groups on the amino acids. Sensing properties of the composites were tested, while pure gold particles were used as the control. The results suggested that the GO/GNPs composites had better linearity and stability in dose-dependent responses to the amino acids than those of the particles, especially in detections for acidic amino acids. Therefore, the nanocomposites platform can provide a convenient and efficient approach for label-free optical detections of important molecules such as amino acids. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Spiked proteomic standard dataset for testing label-free quantitative software and statistical methods.

    Science.gov (United States)

    Ramus, Claire; Hovasse, Agnès; Marcellin, Marlène; Hesse, Anne-Marie; Mouton-Barbosa, Emmanuelle; Bouyssié, David; Vaca, Sebastian; Carapito, Christine; Chaoui, Karima; Bruley, Christophe; Garin, Jérôme; Cianférani, Sarah; Ferro, Myriam; Dorssaeler, Alain Van; Burlet-Schiltz, Odile; Schaeffer, Christine; Couté, Yohann; Gonzalez de Peredo, Anne

    2016-03-01

    This data article describes a controlled, spiked proteomic dataset for which the "ground truth" of variant proteins is known. It is based on the LC-MS analysis of samples composed of a fixed background of yeast lysate and different spiked amounts of the UPS1 mixture of 48 recombinant proteins. It can be used to objectively evaluate bioinformatic pipelines for label-free quantitative analysis, and their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. More specifically, it can be useful for tuning software tools parameters, but also testing new algorithms for label-free quantitative analysis, or for evaluation of downstream statistical methods. The raw MS files can be downloaded from ProteomeXchange with identifier PXD001819. Starting from some raw files of this dataset, we also provide here some processed data obtained through various bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold) in different workflows, to exemplify the use of such data in the context of software benchmarking, as discussed in details in the accompanying manuscript [1]. The experimental design used here for data processing takes advantage of the different spike levels introduced in the samples composing the dataset, and processed data are merged in a single file to facilitate the evaluation and illustration of software tools results for the detection of variant proteins with different absolute expression levels and fold change values.

  10. Whispering Gallery Mode Resonators for Rapid Label-Free Biosensing in Small Volume Droplets

    Science.gov (United States)

    Wildgen, Sarah M.; Dunn, Robert C.

    2015-01-01

    Rapid biosensing requires fast mass transport of the analyte to the surface of the sensing element. To optimize analysis times, both mass transport in solution and the geometry and size of the sensing element need to be considered. Small dielectric spheres, tens of microns in diameter, can act as label-free biosensors using whispering gallery mode (WGM) resonances. WGM resonances are sensitive to the effective refractive index, which changes upon analyte binding to recognition sites on functionalized resonators. The spherical geometry and tens of microns diameter of these resonators provides an efficient target for sensing while their compact size enables detection in limited volumes. Here, we explore conditions leading to rapid analyte detection using WGM resonators as label-free sensors in 10 μL sample droplets. Droplet evaporation leads to potentially useful convective mixing, but also limits the time over which analysis can be completed. We show that active droplet mixing combined with initial binding rate measurements is required for accurate nanomolar protein quantification within the first minute following injection. PMID:25806835

  11. Compact 3D printed module for fluorescence and label-free imaging using evanescent excitation

    Science.gov (United States)

    Pandey, Vikas; Gupta, Shalini; Elangovan, Ravikrishnan

    2018-01-01

    Total internal reflection fluorescence (TIRF) microscopy is widely used for selective excitation and high-resolution imaging of fluorophores, and more recently label-free nanosized objects, with high vertical confinement near a liquid–solid interface. Traditionally, high numerical aperture objectives (>1.4) are used to simultaneously generate evanescent waves and collect fluorescence emission signals which limits their use to small area imaging (3D module called cTIRF that can generate evanescent waves in microscope glass slides via a planar waveguide illumination. The module can be attached as a fixture to any existing optical microscope, converting it into a TIRF and enabling high signal-to-noise ratio (SNR) fluorescence imaging using any magnification objective. As the incidence optics is perpendicular to the detector, label-free evanescent scattering-based imaging of submicron objects can also be performed without using emission filters. SNR is significantly enhanced in this case as compared to cTIRF alone, as seen through our model experiments performed on latex beads and mammalian cells. Extreme flexibility and the low cost of our approach makes it scalable for limited resource settings.

  12. Automated selected reaction monitoring software for accurate label-free protein quantification.

    Science.gov (United States)

    Teleman, Johan; Karlsson, Christofer; Waldemarson, Sofia; Hansson, Karin; James, Peter; Malmström, Johan; Levander, Fredrik

    2012-07-06

    Selected reaction monitoring (SRM) is a mass spectrometry method with documented ability to quantify proteins accurately and reproducibly using labeled reference peptides. However, the use of labeled reference peptides becomes impractical if large numbers of peptides are targeted and when high flexibility is desired when selecting peptides. We have developed a label-free quantitative SRM workflow that relies on a new automated algorithm, Anubis, for accurate peak detection. Anubis efficiently removes interfering signals from contaminating peptides to estimate the true signal of the targeted peptides. We evaluated the algorithm on a published multisite data set and achieved results in line with manual data analysis. In complex peptide mixtures from whole proteome digests of Streptococcus pyogenes we achieved a technical variability across the entire proteome abundance range of 6.5-19.2%, which was considerably below the total variation across biological samples. Our results show that the label-free SRM workflow with automated data analysis is feasible for large-scale biological studies, opening up new possibilities for quantitative proteomics and systems biology.

  13. Label-free in vivo imaging of human leukocytes using two-photon excited endogenous fluorescence

    Science.gov (United States)

    Zeng, Yan; Yan, Bo; Sun, Qiqi; Teh, Seng Khoon; Zhang, Wei; Wen, Zilong; Qu, Jianan Y.

    2013-04-01

    We demonstrate that two-photon excited endogenous fluorescence enables label-free morphological and functional imaging of various human blood cells. Specifically, we achieved distinctive morphological contrast to visualize morphology of important leukocytes, such as polymorphonuclear structure of granulocyte and mononuclear feature of agranulocyte, through the employment of the reduced nicotinamide adenine dinucleotide (NADH) fluorescence signals. In addition, NADH fluorescence images clearly reveal the morphological transformation process of neutrophils during disease-causing bacterial infection. Our findings also show that time-resolved NADH fluorescence can be potentially used for functional imaging of the phagocytosis of pathogens by leukocytes (neutrophils) in vivo. In particular, we found that free-to-bound NADH ratios measured in infected neutrophils increased significantly, which is consistent with a previous study that the energy consumed in the phagocytosis of neutrophils is mainly generated through the glycolysis pathway that leads to the accumulation of free NADH. Future work will focus on further developing and applying label-free imaging technology to investigate leukocyte-related diseases and disorders.

  14. Label-free electrochemical DNA sensor using "click"-functionalized PEDOT electrodes.

    Science.gov (United States)

    Galán, Teresa; Prieto-Simón, Beatriz; Alvira, Margarita; Eritja, Ramón; Götz, Günther; Bäuerle, Peter; Samitier, Josep

    2015-12-15

    Here we describe a label-free electrochemical DNA sensor based on poly(3,4-ethylenedioxythiophene)-modified (PEDOT-modified) electrodes. An acetylene-terminated DNA probe, complementary to a specific "Hepatitis C" virus sequence, was immobilized onto azido-derivatized conducting PEDOT electrodes using "click" chemistry. DNA hybridization was then detected by differential pulse voltammetry, evaluating the changes in the electrochemical properties of the polymer produced by the recognition event. A limit of detection of 0.13 nM was achieved using this highly selective PEDOT-based genosensor, without the need for labeling techniques or microelectrode fabrication processes. These results are promising for the development of label-free and reagentless DNA hybridization sensors based on conducting polymeric substrates. Biosensors can be easily prepared using any DNA sequence containing an alkyne moiety. The data presented here reveal the potential of this DNA sensor for diagnostic applications in the screening of diseases, such as "Hepatitis C", and genetic mutations. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. 3D label-free prostate specific antigen (PSA) immunosensor based on graphene-gold composites.

    Science.gov (United States)

    Jang, Hee Dong; Kim, Sun Kyung; Chang, Hankwon; Choi, Jeong-Woo

    2015-01-15

    Highly sensitive and label-free detection of the prostate specific antigen (PSA) remains a challenge in the diagnosis of prostate cancer. Here, a novel three-dimensional (3D) electrochemical immunosensor capable of sensitive and label-free detection of PSA is reported. This unique immunosensor is equipped with a highly conductive graphene (GR)-based gold (Au) composite modified electrode. The GR-based Au composite is prepared using aerosol spray pyrolysis and the morphology of the composite is the shape of a crumpled GR ball decorated with Au nanoparticles. Unlike the previous research, this novel 3D immunosensor functions very well over a broad linear range of 0-10 ng/mL with a low detection limit of 0.59 ng/mL; furthermore, it exhibits a significantly increased electron transfer and high sensitivity toward PSA. The highest rate of current change with respect to the PSA concentration is 5 μA/(ng/mL). Satisfactory selectivity, reproducibility, and stability of the 3D immunosensor are also exhibited. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Label-free in situ imaging of oil body dynamics and chemistry in germination.

    Science.gov (United States)

    Waschatko, Gustav; Billecke, Nils; Schwendy, Sascha; Jaurich, Henriette; Bonn, Mischa; Vilgis, Thomas A; Parekh, Sapun H

    2016-10-01

    Plant oleosomes are uniquely emulsified lipid reservoirs that serve as the primary energy source during seed germination. These oil bodies undergo significant changes regarding their size, composition and structure during normal seedling development; however, a detailed characterization of these oil body dynamics, which critically affect oil body extractability and nutritional value, has remained challenging because of a limited ability to monitor oil body location and composition during germination in situ Here, we demonstrate via in situ, label-free imaging that oil bodies are highly dynamic intracellular organelles that are morphologically and biochemically remodelled extensively during germination. Label-free, coherent Raman microscopy (CRM) combined with bulk biochemical measurements revealed the temporal and spatial regulation of oil bodies in native soya bean cotyledons during the first eight days of germination. Oil bodies undergo a cycle of growth and shrinkage that is paralleled by lipid and protein compositional changes. Specifically, the total protein concentration associated with oil bodies increases in the first phase of germination and subsequently decreases. Lipids contained within the oil bodies change in saturation and chain length during germination. Our results show that CRM is a well-suited platform to monitor in situ lipid dynamics and local chemistry and that oil bodies are actively remodelled during germination. This underscores the dynamic role of lipid reservoirs in plant development. © 2016 The Authors.

  17. Optical Microfibre Based Photonic Components and Their Applications in Label-Free Biosensing.

    Science.gov (United States)

    Wang, Pengfei; Bo, Lin; Semenova, Yuliya; Farrell, Gerald; Brambilla, Gilberto

    2015-07-22

    Optical microfibre photonic components offer a variety of enabling properties, including large evanescent fields, flexibility, configurability, high confinement, robustness and compactness. These unique features have been exploited in a range of applications such as telecommunication, sensing, optical manipulation and high Q resonators. Optical microfibre biosensors, as a class of fibre optic biosensors which rely on small geometries to expose the evanescent field to interact with samples, have been widely investigated. Due to their unique properties, such as fast response, functionalization, strong confinement, configurability, flexibility, compact size, low cost, robustness, ease of miniaturization, large evanescent field and label-free operation, optical microfibres based biosensors seem a promising alternative to traditional immunological methods for biomolecule measurements. Unlabeled DNA and protein targets can be detected by monitoring the changes of various optical transduction mechanisms, such as refractive index, absorption and surface plasmon resonance, since a target molecule is capable of binding to an immobilized optical microfibre. In this review, we critically summarize accomplishments of past optical microfibre label-free biosensors, identify areas for future research and provide a detailed account of the studies conducted to date for biomolecules detection using optical microfibres.

  18. Rapid and label-free separation of Burkitt's lymphoma cells from red blood cells by optically-induced electrokinetics.

    Directory of Open Access Journals (Sweden)

    Wenfeng Liang

    Full Text Available Early stage detection of lymphoma cells is invaluable for providing reliable prognosis to patients. However, the purity of lymphoma cells in extracted samples from human patients' marrow is typically low. To address this issue, we report here our work on using optically-induced dielectrophoresis (ODEP force to rapidly purify Raji cells' (a type of Burkitt's lymphoma cell sample from red blood cells (RBCs with a label-free process. This method utilizes dynamically moving virtual electrodes to induce negative ODEP force of varying magnitudes on the Raji cells and RBCs in an optically-induced electrokinetics (OEK chip. Polarization models for the two types of cells that reflect their discriminate electrical properties were established. Then, the cells' differential velocities caused by a specific ODEP force field were obtained by a finite element simulation model, thereby established the theoretical basis that the two types of cells could be separated using an ODEP force field. To ensure that the ODEP force dominated the separation process, a comparison of the ODEP force with other significant electrokinetics forces was conducted using numerical results. Furthermore, the performance of the ODEP-based approach for separating Raji cells from RBCs was experimentally investigated. The results showed that these two types of cells, with different concentration ratios, could be separated rapidly using externally-applied electrical field at a driven frequency of 50 kHz at 20 Vpp. In addition, we have found that in order to facilitate ODEP-based cell separation, Raji cells' adhesion to the OEK chip's substrate should be minimized. This paper also presents our experimental results of finding the appropriate bovine serum albumin concentration in an isotonic solution to reduce cell adhesion, while maintaining suitable medium conductivity for electrokinetics-based cell separation. In short, we have demonstrated that OEK technology could be a promising tool for

  19. Raman-Activated Droplet Sorting (RADS) for Label-Free High-Throughput Screening of Microalgal Single-Cells.

    Science.gov (United States)

    Wang, Xixian; Ren, Lihui; Su, Yetian; Ji, Yuetong; Liu, Yaoping; Li, Chunyu; Li, Xunrong; Zhang, Yi; Wang, Wei; Hu, Qiang; Han, Danxiang; Xu, Jian; Ma, Bo

    2017-11-21

    Raman-activated cell sorting (RACS) has attracted increasing interest, yet throughput remains one major factor limiting its broader application. Here we present an integrated Raman-activated droplet sorting (RADS) microfluidic system for functional screening of live cells in a label-free and high-throughput manner, by employing AXT-synthetic industrial microalga Haematococcus pluvialis (H. pluvialis) as a model. Raman microspectroscopy analysis of individual cells is carried out prior to their microdroplet encapsulation, which is then directly coupled to DEP-based droplet sorting. To validate the system, H. pluvialis cells containing different levels of AXT were mixed and underwent RADS. Those AXT-hyperproducing cells were sorted with an accuracy of 98.3%, an enrichment ratio of eight folds, and a throughput of ∼260 cells/min. Of the RADS-sorted cells, 92.7% remained alive and able to proliferate, which is equivalent to the unsorted cells. Thus, the RADS achieves a much higher throughput than existing RACS systems, preserves the vitality of cells, and facilitates seamless coupling with downstream manipulations such as single-cell sequencing and cultivation.

  20. Label-free electrochemical immunosensor based on Nile blue A-reduced graphene oxide nanocomposites for carcinoembryonic antigen detection.

    Science.gov (United States)

    Gao, Yan-Sha; Zhu, Xiao-Fei; Xu, Jing-Kun; Lu, Li-Min; Wang, Wen-Min; Yang, Tao-Tao; Xing, Hua-Kun; Yu, Yong-Fang

    2016-05-01

    In this article, a novel, label-free, and inherent electroactive redox immunosensor for carcinoembryonic antigen (CEA) based on gold nanoparticles (AuNPs) and Nile blue A (NB) hybridized electrochemically reduced graphene oxide (NB-ERGO) is proposed. The composite of NB-graphene oxide (NB-GO) was prepared by π-π stacking interaction. Then, chronoamperometry was adopted to simultaneously reduce HAuCl4 and nanocomposites of NB-GO for synthesizing AuNPs/NB-ERGO. The immunosensor was fabricated by capturing CEA antibody (anti-CEA) at this nanocomposite modified electrode. The immunosensor determination was based on the fact that, due to the formation of antigen-antibody immunocomplex, the decreased response currents of NB were directly proportional to the concentrations of CEA. Under optimal conditions, the linear range of the proposed immunosensor was estimated to be from 0.001 to 40 ng ml(-1) and the detection limit was estimated to be 0.00045 ng ml(-1). The proposed immunosensor was used to determine CEA in clinical serum samples with satisfactory results. The proposed method may provide promising potential application in clinical immunoassays with the properties of facile procedure, stability, high sensitivity, and selectivity. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Label-free detection of bovine serum albumin based on an in-fiber Mach-Zehnder interferometric biosensor.

    Science.gov (United States)

    Li, Zhengyong; Liao, Changrui; Chen, Danni; Song, Jun; Jin, Wei; Peng, Gang-Ding; Zhu, Feng; Wang, Ying; He, Jun; Wang, Yiping

    2017-07-24

    We propose and experimentally verify an innovative label-free optical fiber biosensor based on a Mach-Zehnder interferometer for bovine serum albumin (BSA) concentration detection. The proposed fiber biosensor utilized a micro-cavity within a single-mode fiber to induce Mach-Zehnder interference. A remarkable feature of this biosensor is that external media can directly interact with the fiber core signal through microfluidic channels connected to the micro-cavity and sensor surface. The device was fabricated by means of femtosecond laser micromachining and chemical etching. A fiber interferometer of this type exhibits an ultrahigh refractive index sensitivity of -10,055 nm/RIU and a detection limit of 3.5 × 10 -5 RIU. Different concentrations of BSA with an infinitesimally small refractive index difference can be clearly differentiated in situ by the interferential spectra of the structure. Experiments demonstrated the biosensor exhibited a BSA solution concentration sensitivity of -38.9 nm/(mg/mL) and a detection limit of 2.57 × 10 -4 mg/mL, respectively. Moreover, this biosensor is a sub-microliter dose and ultrasensitive at the low concentrations detected in BSA, which make it a promising for biochemical applications such as DNA hybridization, cancer screenings, medicine examination and environmental engineering, etc.

  2. Label-free tracking of single extracellular vesicles in a nano-fluidic optical fiber (Conference Presentation)

    Science.gov (United States)

    van der Pol, Edwin; Weidlich, Stefan; Lahini, Yoav; Coumans, Frank A. W.; Sturk, Auguste; Nieuwland, Rienk; Schmidt, Markus A.; Faez, Sanli; van Leeuwen, Ton G.

    2016-03-01

    Background: Extracellular vesicles, such as exosomes, are abundantly present in human body fluids. Since the size, concentration and composition of these vesicles change during disease, vesicles have promising clinical applications, including cancer diagnosis. However, since ~70% of the vesicles have a diameter vesicles remains challenging. Thus far, vesicles vesicles to be adhered to a surface. Consequently, the majority of vesicles have never been studied in their physiological environment. We present a novel label-free optical technique to track single vesicles vesicles were contained within a single-mode light-guiding silica fiber containing a 600 nm nano-fluidic channel. Light from a diode laser (660 nm wavelength) was coupled to the fiber, resulting in a strongly confined optical mode in the nano-fluidic channel, which continuously illuminated the freely diffusing vesicles inside the channel. The elastic light scattering from the vesicles, in the direction orthogonal to the fiber axis, was collected using a microscope objective (NA=0.95) and imaged with a home-built microscope. Results: We have tracked single urinary vesicles as small as 35 nm by elastic light scattering. Please note that vesicles are low-refractive index (nvesicles vesicle-based clinical applications.

  3. Direct observation of DNA translocation influenced by electrically gated nanopores

    Science.gov (United States)

    Ando, Genki; Moriya, Hiroki; Tsukahira, Kenta; Yano, Satoshi; Mitsui, Toshiyuki

    2012-02-01

    One of remarkable recent developments in the solid state nanopore based DNA analysis is adding the ability to control electric potential near nanopore as a gate electrode by patterning metal in or on nanopore. In this approach, better control of DNA translocations for example, slowing down the translocation speed might be expected. We have fabricated insulator-metal-insulator nanopores of rather large 100 nm pore in diameter. The 100 nm diameter pores allow us to observe the translocation of lambda-DNA molecules directly by means of fluorescence microscopy without heavy clogging of the DNA molecules into the pores. By controlling ?gate voltage? on metal relative to the cis and trans voltages, the translocation rates of DNA are able to change. Interestingly, applying pulse voltage to the gate metal near 100 ms to reverse the direction of the electric field near the cis side of nanopore reverses the direction of the DNA translocation instantaneously. This in fact provides us a new way to repeat translocation of the same DNA molecule. Furthermore, repeating the pulse tends to clear off the clogged DNA molecules in nanopore. We will present more details of these phenomena caused by the gate voltages.

  4. Carbon nanotubes-based label-free affinity sensors for environmental monitoring.

    Science.gov (United States)

    Sarkar, Tapan; Gao, Yingning; Mulchandani, Ashok

    2013-07-01

    Nanostructures, such as nanowires, nanobelts, nanosprings, and nanotubes, are receiving growing interest as transducer elements of bio/chemical sensors as they provide high sensitivity, multiplexing, small size, and portability. Single-walled carbon nanotubes (SWNTs) are one such class of nanostructure materials that exhibit superior sensing behavior due to its large-surface carbon atoms that are highly responsive to surface adsorption events. Further, their compatibility with modern microfabrication technologies and facile functionalization with molecular recognition elements make them promising candidates for bio/chemical sensors applications. Here, we review recent results on nanosensors based on SWNTs modified with biological receptors such as aptamers, antibodies, and binding proteins, to develop highly sensitive, selective, rapid, and cost-effective label-free chemiresistor/field-effect transistor nanobiosensors for applications in environmental monitoring.

  5. Label-free in vivo imaging of peripheral nerve by multispectral photoacoustic tomography.

    Science.gov (United States)

    Li, Rui; Phillips, Evan; Wang, Pu; Goergen, Craig J; Cheng, Ji-Xin

    2016-01-01

    Unintentional surgical damage to nerves is mainly due to poor visualization of nerve tissue relative to adjacent structures. Multispectral photoacoustic tomography can provide chemical information with specificity and ultrasonic spatial resolution with centimeter imaging depth, making it a potential tool for noninvasive neural imaging. To implement this label-free imaging approach, a multispectral photoacoustic tomography platform was built. Imaging depth and spatial resolution were characterized. In vivo imaging of the femoral nerve that is 2 mm deep in a nude mouse was performed. Through multivariate curve resolution analysis, the femoral nerve was discriminated from the femoral artery and chemical maps of their spatial distributions were generated. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Label-Free Imaging and Biochemical Characterization of Bovine Sperm Cells

    Directory of Open Access Journals (Sweden)

    Maria Antonietta Ferrara

    2015-04-01

    Full Text Available A full label-free morphological and biochemical characterization is desirable to select spermatozoa during preparation for artificial insemination. In order to study these fundamental parameters, we take advantage of two attractive techniques: digital holography (DH and Raman spectroscopy (RS. DH presents new opportunities for studying morphological aspect of cells and tissues non-invasively, quantitatively and without the need for staining or tagging, while RS is a very specific technique allowing the biochemical analysis of cellular components with a spatial resolution in the sub-micrometer range. In this paper, morphological and biochemical bovine sperm cell alterations were studied using these techniques. In addition, a complementary DH and RS study was performed to identify X- and Y-chromosome-bearing sperm cells. We demonstrate that the two techniques together are a powerful and highly efficient tool elucidating some important criterions for sperm morphological selection and sex-identification, overcoming many of the limitations associated with existing protocols.

  7. Design of a Label-Free, Distributed Bragg Grating Resonator Based Dielectric Waveguide Biosensor

    Directory of Open Access Journals (Sweden)

    Florian Kehl

    2015-01-01

    Full Text Available In this work, we present a resonant, dielectric waveguide device based on distributed Bragg gratings for label-free biosensing applications. The refractive index sensitive optical transducer aims at improving the performance of planar waveguide grating sensor systems with limited Q-factor and dynamic range by combing the advantages of resonant cavities, such as a multitude of resonance peaks with high finesse, with the manageable complexity of waveguide grating couplers. The general sensor concept is introduced and supported by theoretical considerations as well as numerical simulations based on Coupled Mode Theory. In contrast to a single Bragg grating reflector, the presented Fabry-Pérot type distributed Bragg resonator exhibits an extended measurement range as well as relaxed fabrication tolerances. The resulting, relatively simple sensor structure can be fabricated with standard lithographic means and is independent of expensive light-sources and/or detectors, making an affordable but sensitive device, potentially suitable for point-of-care applications.

  8. Nanostructured Electrochemical Biosensors for Label-Free Detection of Water- and Food-Borne Pathogens.

    Science.gov (United States)

    Reta, Nekane; Saint, Christopher P; Michelmore, Andrew; Prieto-Simon, Beatriz; Voelcker, Nicolas H

    2018-02-21

    The emergence of nanostructured materials has opened new horizons in the development of next generation biosensors. Being able to control the design of the electrode interface at the nanoscale combined with the intrinsic characteristics of the nanomaterials engenders novel biosensing platforms with improved capabilities. The purpose of this review is to provide a comprehensive and critical overview of the latest trends in emerging nanostructured electrochemical biosensors. A detailed description and discussion of recent approaches to construct label-free electrochemical nanostructured electrodes is given with special focus on pathogen detection for environmental monitoring and food safety. This includes the use of nanoscale materials such as nanotubes, nanowires, nanoparticles, and nanosheets as well as porous nanostructured materials including nanoporous anodic alumina, mesoporous silica, porous silicon, and polystyrene nanochannels. These platforms may pave the way toward the development of point-of-care portable electronic devices for applications ranging from environmental analysis to biomedical diagnostics.

  9. Label-free brainwide visualization of senile plaque using cryo-micro-optical sectioning tomography.

    Science.gov (United States)

    Luo, Yilin; Wang, Anle; Liu, Mengmeng; Lei, Tian; Zhang, Xiaochuan; Gao, Zhaobing; Jiang, Hualiang; Gong, Hui; Yuan, Jing

    2017-11-01

    Optical visualization of pathological changes in Alzheimer's disease (AD) can facilitate exploration of disease mechanisms and treatments. However, existing optical imaging methods have limitations on mapping pathological evolution in the whole mouse brain. Previous research indicated endogenous fluorescence contrast of senile plaques. Therefore, we develop cryo-micro-optical sectioning tomography (cryo-MOST) to capture intrinsic fluorescence distribution of senile plaques at a micrometer-level resolution in the whole brain. Validation using immunofluorescence demonstrates the capacity of cryo-MOST to visualize and distinguish senile plaques with competent sensitivity and spatial resolution. Compared with imaging in room temperature, cryo-MOST provides better signal intensity and signal-to-noise ratio. Using cryo-MOST, we obtained whole-brain coronal distribution of senile plaques in a transgenic mouse without exogenous dye. Capable of label-free brainwide visualization of Alzheimer's pathology, cryo-MOST may be potentially useful for understanding neurodegenerative disease mechanisms and evaluating drug efficacy.

  10. Asynchronous Magnetic Bead Rotation (AMBR Microviscometer for Label-Free DNA Analysis

    Directory of Open Access Journals (Sweden)

    Yunzi Li

    2014-03-01

    Full Text Available We have developed a label-free viscosity-based DNA detection system, using paramagnetic beads as an asynchronous magnetic bead rotation (AMBR microviscometer. We have demonstrated experimentally that the bead rotation period is linearly proportional to the viscosity of a DNA solution surrounding the paramagnetic bead, as expected theoretically. Simple optical measurement of asynchronous microbead motion determines solution viscosity precisely in microscale volumes, thus allowing an estimate of DNA concentration or average fragment length. The response of the AMBR microviscometer yields reproducible measurement of DNA solutions, enzymatic digestion reactions, and PCR systems at template concentrations across a 5000-fold range. The results demonstrate the feasibility of viscosity-based DNA detection using AMBR in microscale aqueous volumes.

  11. Biosensor for label-free DNA quantification based on functionalized LPGs.

    Science.gov (United States)

    Gonçalves, Helena M R; Moreira, Luis; Pereira, Leonor; Jorge, Pedro; Gouveia, Carlos; Martins-Lopes, Paula; Fernandes, José R A

    2016-10-15

    A label-free fiber optic biosensor based on a long period grating (LPG) and a basic optical interrogation scheme using off the shelf components is used for the detection of in-situ DNA hybridization. A new methodology is proposed for the determination of the spectral position of the LPG mode resonance. The experimental limit of detection obtained for the DNA was 62±2nM and the limit of quantification was 209±7nM. The sample specificity was experimentally demonstrated using DNA targets with different base mismatches relatively to the probe and was found that the system has a single base mismatch selectivity. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Hydrodynamic and label-free sorting of circulating tumor cells from whole blood

    Science.gov (United States)

    Geislinger, Thomas M.; Stamp, Melanie E. M.; Wixforth, Achim; Franke, Thomas

    2015-11-01

    We demonstrate continuous, passive, and label-free sorting of different in vitro cancer cell lines (MV3, MCF7, and HEPG2) as model systems for circulating tumor cells (CTCs) from undiluted whole blood employing the non-inertial lift effect as driving force. This purely viscous, repulsive cell-wall interaction is sensitive to cell size and deformability differences and yields highly efficient cell separation and high enrichment factors. We show that the performance of the device is robust over a large range of blood cell concentrations and flow rates as well as for the different cell lines. The collected samples usually contain more than 90% of the initially injected CTCs and exhibit average enrichment factors of more than 20 for sorting from whole blood samples.

  13. Label-free Electrochemical Impedance Detection of C-reactive Protein

    Science.gov (United States)

    Whited, Allison; Singh, K.. V.; Solanki, Raj

    2009-05-01

    C-reactive protein, CRP, is a marker present in human serum indicating inflammation and infection. By measuring the amount present in serum, it is possible to monitor the effectiveness of a treatment or roughly gauge the risk of heart disease. Using a double antibody capture system immobilized on an interdigitated electrode array, a label-free device was developed to detect the presence of CRP present in buffer solution and various concentrations of human serum. Electrochemical impedance spectroscopy was used to measure the end point data of the binding signal as the assay was exposed to varying amounts of CRP in the presence of a constant concentration of anti-CRP. The sensor is able to achieve linear detection in both buffer solution and human serum spiked with CRP in the range of 1ng/ml to 1ug/ml. The sensor developed can be integrated into a portable microfluidic device.

  14. Label-free, multiplexed detection of bacterial tmRNA using silicon photonic microring resonators.

    Science.gov (United States)

    Scheler, Ott; Kindt, Jared T; Qavi, Abraham J; Kaplinski, Lauris; Glynn, Barry; Barry, Thomas; Kurg, Ants; Bailey, Ryan C

    2012-01-01

    A label-free biosensing method for the sensitive detection and identification of bacterial transfer-messenger RNA (tmRNA) is presented employing arrays of silicon photonic microring resonators. Species specific tmRNA molecules are targeted by complementary DNA capture probes that are covalently attached to the sensor surface. Specific hybridization is monitored in near real-time by observing the resonance wavelength shift of each individual microring. The sensitivity of the biosensing platform allowed for detection down to 53 fmol of Streptococcus pneumoniae tmRNA, equivalent to approximately 3.16×10(7) CFU of bacteria. The simplicity and scalability of this biosensing approach makes it a promising tool for the rapid identification of different bacteria via tmRNA profiling. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Portable microfluidic raman system for rapid, label-free early disease signature detection

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Meiye [Sandia National Laboratories (SNL-CA), Livermore, CA (United States); Davis, Ryan Wesley [Sandia National Laboratories (SNL-CA), Livermore, CA (United States); Hatch, Anson [Sandia National Laboratories (SNL-CA), Livermore, CA (United States)

    2015-09-01

    In the early stages of infection, patients develop non-specific or no symptoms at all. While waiting for identification of the infectious agent, precious window of opportunity for early intervention is lost. The standard diagnostics require affinity reagents and sufficient pathogen titers to reach the limit of detection. In the event of a disease outbreak, triaging the at-risk population rapidly and reliably for quarantine and countermeasure is more important than the identification of the pathogen by name. To expand Sandia's portfolio of Biological threat management capabilities, we will utilize Raman spectrometry to analyze immune subsets in whole blood to rapidly distinguish infected from non-infected, and bacterial from viral infection, for the purpose of triage during an emergency outbreak. The goal of this one year LDRD is to determine whether Raman spectroscopy can provide label-free detection of early disease signatures, and define a miniaturized Raman detection system meeting requirements for low- resource settings.

  16. Improvements in low-cost label-free QPI microscope for live cell imaging

    Science.gov (United States)

    Seniya, C.; Towers, C. E.; Towers, D. P.

    2017-07-01

    This paper reports an improvement in the development of a low-cost QPI microscope offering new capabilities in term of phase measurement accuracy for label-free live samples in the longer term (i.e., hours to days). The spatially separated scattered and non-scattered image light fields are reshaped in the Fourier plane and modulated to form an interference image at a CCD camera. The apertures that enable these two beams to be generated have been optimised by means of laser-cut apertures placed on the mirrors of a Michelson interferometer and has improved the phase measuring and reconstruction capability of the QPI microscope. The microscope was tested with transparent onion cells as an object of interest.

  17. Development and Bioanalytical Applications of a White Light Reflectance Spectroscopy Label-Free Sensing Platform

    Directory of Open Access Journals (Sweden)

    Georgios Koukouvinos

    2017-10-01

    Full Text Available The development of a sensing platform based on white light reflectance spectroscopy (WLRS is presented. The evolution of the system, from polymer film characterization and sensing of volatile organic compounds to biosensor for the label-free determination of either high (e.g., proteins or low molecular weight analytes (e.g., pesticides, is described. At the same time, the passage from single to multi-analyte determinations, and from a laboratory prototype set-up to a compact device appropriate for on-site determination, is outlined. The improvements made on both the sensor and the optical set-up, and the concomitant advances in the analytical characteristics and the robustness of the assays performed with the different layouts, are also presented. Finally, the future perspectives of the system, aiming for the creation of a standalone instrument to be used by non-experts, will be discussed.

  18. Label free detection of 5′ hydroxymethylcytosine within CpG Islands using optical sensors

    Science.gov (United States)

    Hawk, Rasheeda M.; Armani, Andrea M.

    2014-01-01

    Significant research has been invested in correlating genetic variations with different disease probabilities. Recently, it has become apparent that other DNA modifications, such as the addition of a methyl or hydroxymethyl group to cytosine, can also play a role. While these modifications do not change the sequence, they can negatively impact the function. Therefore, it is critical to be able to both read the genetic code and identify these modifications. Currently, the detection of hydroxymethylated cytosine (5′hmC) and the two closely related variants, cytosine (C) and 5′methylcytosine (5′mC), relies on a combination of nucleotide modification steps, followed by PCR and gene sequencing. However, this approach is not ideal because transcription errors which are inherent to the PCR process can be misinterpreted as fluctuations in the relative C:5′mC:5′hmC concentrations. As such, an alternative method which does not rely on PCR or nucleotide modification is desirable. One approach is based on label-free optical resonant cavity sensors. In the present work, toroidal resonant cavity sensors are functionalized with antibodies to enable label-free detection and discrimination between C, 5′mC, and 5′hmC in real-time without PCR. Specifically, epoxide chemistry is used to covalently attach the 5′hmC antibody to the surface of the cavity. Subsequently, to thoroughly characterize the sensor platform, detection of C, 5′mC, and 5′hmC is performed over a concentration range from pM to nM. At low (pM) concentrations, the hydroxymethylated cytosine produces a significantly larger signal than the structurally similar epigenetic markers; thus demonstrating the applicability of this platform. PMID:25461158

  19. Novel label-free biosensing technology for monitoring of aqueous solutions (Conference Presentation)

    Science.gov (United States)

    Kehl, Florian; Bielecki, Robert; Follonier, Stephane; Dorokhin, Denis

    2016-03-01

    Waste water, drinking water and other industrial water sources are more and more/increasingly polluted with a large variety of contaminants, such as pesticides or residuals of pharmaceuticals. These compounds can impact human and animal organisms and lead to serious health issues. Today, in order to analyze the presence and quantity of the abovementioned micropollutants, samples are typically sent to specialized centralized laboratories and their processing may take up to several days. In order to meet the demand for continuous and consistent monitoring of aqueous solutions we propose a novel label-free technology system comprising proprietary chip and reader device designs. The core of the system is constituted by a planar-grated-waveguide (PGW) chip. Label-free biosensors, based on PGWs are sensitive to effective refractive index changes caused by the adsorption of biomolecules (micropollutants) onto the sensor surface or due to refractive index changes of the bulk solution. The presented reader device operates with a novel readout concept based on a scanning MEMS mirror for the angular interrogation of input grating couplers at a high repetition rate. The reader has fully integrated optics, electronics and fluidics and at the same time consumes limited energy (portable, field use ready). In the recent experiments, the effectiveness of the technology has been demonstrated with various liquids and bioassays showing (i) an excellent refractometric sensitivity with a limit of detection towards effective refractive index changes of ▵neff < 2 x 10-7, and (ii) the capability to perform affinity measurements for large (<150 kDa) and small (<250 Da) molecules.

  20. Old tree with new shoots: silver nanoparticles for label-free and colorimetric mercury ions detection

    Energy Technology Data Exchange (ETDEWEB)

    Gao Shuyan, E-mail: shuyangao@htu.cn; Jia Xiaoxia; Chen Yanli [Henan Normal University, College of Chemistry and Environmental Science (China)

    2013-01-15

    Mercury in the environment from global mercury emissions as well as various forms of contamination poses severe threats to both human health and the environment. Long-term exposure to high levels of Hg-based toxins results in serious and irreversible damage of the central nervous system and other organs. Therefore, the development of effective sensing systems for mercury detection becomes an increasing demand. In this article, a yogurt-mediated silver nanostructure is reported to be unprecedentedly used in the naked-eye and label-free detection of mercury. The method relies on the redox reaction resulting from the electrode potential difference between Ag{sup +}/Ag (0.7996 V) and Hg{sup 2+}/Hg{sub 2}{sup 2+} (0.920 V) that makes colorless Hg{sup 2+} ions which oxidize colored silver nanoparticle (AgNP) to colorless Ag+. The labor-intensive modification of AgNPs and expensive labeling are avoided, and the traditional AuNPs are substituted by AgNPs in this Hg{sup 2+} ions sensing platform, which makes it facile, low-cost, and particularly useful for home, clinic, or field applications as well as resource-limited conditions. This sensing system achieves a detection limit as low as 10 nM, lower than the toxicity level of Hg{sup 2+} ions in drinking water (30 nM) defined by World Health Organization, and exhibits excellent selectivity, largely free from the matrix effect of the real water samples. This visual label-free Hg{sup 2+} ions sensing motif shows great promise for sensing Hg{sup 2+} ions in terms of sensitivity, selectivity, cost, and maneuverability. It is also a good example for the organic combination of green chemistry and functional materials, which may trigger interest in furthering biosystems for environmental science applications.

  1. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    Energy Technology Data Exchange (ETDEWEB)

    Tam, Phuong Dinh, E-mail: phuongdinhtam@gmail.com; Thang, Cao Xuan, E-mail: thang.caoxuan@hust.edu.vn

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO{sub 2} nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO{sub 2} nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) {sub 6}] {sup 3−/4−} as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL{sup −1}. Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10{sup 2} CFU/mL to 1.0 × 10{sup 4} CFU/mL. • The detection limit of the immunosensor was 1.0 × 10{sup 2} CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL{sup −1}.

  2. Label-free electrochemical immunosensor based on cerium oxide nanowires for Vibrio cholerae O1 detection

    International Nuclear Information System (INIS)

    Tam, Phuong Dinh; Thang, Cao Xuan

    2016-01-01

    This paper developed a label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application. The CeO 2 nanowires were synthesized by hydrothermal reaction. The immobilization of Anti-V. cholerae O1 onto CeO 2 nanowire-deposited sensor was performed via an amino ester, which was created by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and sulfo-N-hydroxysuccinimide. The electrochemical responses of the immunosensor were studied by electrochemical impedance spectroscopy with [Fe (CN) 6 ] 3−/4− as redox probe. A linear response in electron transfer resistance for cell of V. cholerae O1 concentration was found in the range of 1.0 × 10 2 CFU/mL to 1.0 × 10 4 CFU/mL. The detection limit of the immunosensor was 1.0 × 10 2 CFU/mL. The immunosensor sensitivity was 56.82 Ω/CFU·mL −1 . Furthermore, the parameters affecting immunosensor response were also investigated, as follows: pH value, immunoreaction time, incubation temperature, and anti-V. cholerae O1 concentration. - Highlights: • A label-free immunosensor based on cerium oxide nanowire for Vibrio cholerae O1 detection application was developed. • A linear response was found in the range of 1.0 × 10 2 CFU/mL to 1.0 × 10 4 CFU/mL. • The detection limit of the immunosensor was 1.0 × 10 2 CFU/mL. • The immunosensor sensitivity was 56.82 Ω/CFU.mL −1 .

  3. A comprehensive evaluation of popular proteomics software workflows for label-free proteome quantification and imputation.

    Science.gov (United States)

    Välikangas, Tommi; Suomi, Tomi; Elo, Laura L

    2017-05-31

    Label-free mass spectrometry (MS) has developed into an important tool applied in various fields of biological and life sciences. Several software exist to process the raw MS data into quantified protein abundances, including open source and commercial solutions. Each software includes a set of unique algorithms for different tasks of the MS data processing workflow. While many of these algorithms have been compared separately, a thorough and systematic evaluation of their overall performance is missing. Moreover, systematic information is lacking about the amount of missing values produced by the different proteomics software and the capabilities of different data imputation methods to account for them.In this study, we evaluated the performance of five popular quantitative label-free proteomics software workflows using four different spike-in data sets. Our extensive testing included the number of proteins quantified and the number of missing values produced by each workflow, the accuracy of detecting differential expression and logarithmic fold change and the effect of different imputation and filtering methods on the differential expression results. We found that the Progenesis software performed consistently well in the differential expression analysis and produced few missing values. The missing values produced by the other software decreased their performance, but this difference could be mitigated using proper data filtering or imputation methods. Among the imputation methods, we found that the local least squares (lls) regression imputation consistently increased the performance of the software in the differential expression analysis, and a combination of both data filtering and local least squares imputation increased performance the most in the tested data sets. © The Author 2017. Published by Oxford University Press.

  4. Label-free SnO2 nanowire FET biosensor for protein detection

    Science.gov (United States)

    Jakob, Markus H.; Dong, Bo; Gutsch, Sebastian; Chatelle, Claire; Krishnaraja, Abinaya; Weber, Wilfried; Zacharias, Margit

    2017-06-01

    Novel tin oxide field-effect-transistors (SnO2 NW-FET) for pH and protein detection applicable in the healthcare sector are reported. With a SnO2 NW-FET the proof-of-concept of a bio-sensing device is demonstrated using the carrier transport control of the FET channel by a (bio-) liquid modulated gate. Ultra-thin Al2O3 fabricated by a low temperature atomic layer deposition (ALD) process represents a sensitive layer to H+ ions safeguarding the nanowire at the same time. Successful pH sensitivity is demonstrated for pH ranging from 3 to 10. For protein detection, the SnO2 NW-FET is functionalized with a receptor molecule which specifically interacts with the protein of interest to be detected. The feasibility of this approach is demonstrated via the detection of a biotinylated protein using a NW-FET functionalized with streptavidin. An immediate label-free electronic read-out of the signal is shown. The well-established Enzyme-Linked Immunosorbent Assay (ELISA) method is used to determine the optimal experimental procedure which would enable molecular binding events to occur while being compatible with a final label-free electronic read-out on a NW-FET. Integration of the bottom-up fabricated SnO2 NW-FET pH- and biosensor into a microfluidic system (lab-on-a-chip) allows the automated analysis of small volumes in the 400 μl range as would be desired in portable on-site point-of-care (POC) devices for medical diagnosis.

  5. Old tree with new shoots: silver nanoparticles for label-free and colorimetric mercury ions detection

    Science.gov (United States)

    Gao, Shuyan; Jia, Xiaoxia; Chen, Yanli

    2013-01-01

    Mercury in the environment from global mercury emissions as well as various forms of contamination poses severe threats to both human health and the environment. Long-term exposure to high levels of Hg-based toxins results in serious and irreversible damage of the central nervous system and other organs. Therefore, the development of effective sensing systems for mercury detection becomes an increasing demand. In this article, a yogurt-mediated silver nanostructure is reported to be unprecedentedly used in the naked-eye and label-free detection of mercury. The method relies on the redox reaction resulting from the electrode potential difference between Ag+/Ag (0.7996 V) and Hg2+/Hg2 2+ (0.920 V) that makes colorless Hg2+ ions which oxidize colored silver nanoparticle (AgNP) to colorless Ag+. The labor-intensive modification of AgNPs and expensive labeling are avoided, and the traditional AuNPs are substituted by AgNPs in this Hg2+ ions sensing platform, which makes it facile, low-cost, and particularly useful for home, clinic, or field applications as well as resource-limited conditions. This sensing system achieves a detection limit as low as 10 nM, lower than the toxicity level of Hg2+ ions in drinking water (30 nM) defined by World Health Organization, and exhibits excellent selectivity, largely free from the matrix effect of the real water samples. This visual label-free Hg2+ ions sensing motif shows great promise for sensing Hg2+ ions in terms of sensitivity, selectivity, cost, and maneuverability. It is also a good example for the organic combination of green chemistry and functional materials, which may trigger interest in furthering biosystems for environmental science applications.

  6. Old tree with new shoots: silver nanoparticles for label-free and colorimetric mercury ions detection

    International Nuclear Information System (INIS)

    Gao Shuyan; Jia Xiaoxia; Chen Yanli

    2013-01-01

    Mercury in the environment from global mercury emissions as well as various forms of contamination poses severe threats to both human health and the environment. Long-term exposure to high levels of Hg-based toxins results in serious and irreversible damage of the central nervous system and other organs. Therefore, the development of effective sensing systems for mercury detection becomes an increasing demand. In this article, a yogurt-mediated silver nanostructure is reported to be unprecedentedly used in the naked-eye and label-free detection of mercury. The method relies on the redox reaction resulting from the electrode potential difference between Ag + /Ag (0.7996 V) and Hg 2+ /Hg 2 2+ (0.920 V) that makes colorless Hg 2+ ions which oxidize colored silver nanoparticle (AgNP) to colorless Ag+. The labor-intensive modification of AgNPs and expensive labeling are avoided, and the traditional AuNPs are substituted by AgNPs in this Hg 2+ ions sensing platform, which makes it facile, low-cost, and particularly useful for home, clinic, or field applications as well as resource-limited conditions. This sensing system achieves a detection limit as low as 10 nM, lower than the toxicity level of Hg 2+ ions in drinking water (30 nM) defined by World Health Organization, and exhibits excellent selectivity, largely free from the matrix effect of the real water samples. This visual label-free Hg 2+ ions sensing motif shows great promise for sensing Hg 2+ ions in terms of sensitivity, selectivity, cost, and maneuverability. It is also a good example for the organic combination of green chemistry and functional materials, which may trigger interest in furthering biosystems for environmental science applications.

  7. Resistive graphene humidity sensors with rapid and direct electrical readout

    Science.gov (United States)

    Smith, Anderson D.; Elgammal, Karim; Niklaus, Frank; Delin, Anna; Fischer, Andreas C.; Vaziri, Sam; Forsberg, Fredrik; Råsander, Mikael; Hugosson, Håkan; Bergqvist, Lars; Schröder, Stephan; Kataria, Satender; Östling, Mikael; Lemme, Max C.

    2015-11-01

    We demonstrate humidity sensing using a change of the electrical resistance of single-layer chemical vapor deposited (CVD) graphene that is placed on top of a SiO2 layer on a Si wafer. To investigate the selectivity of the sensor towards the most common constituents in air, its signal response was characterized individually for water vapor (H2O), nitrogen (N2), oxygen (O2), and argon (Ar). In order to assess the humidity sensing effect for a range from 1% relative humidity (RH) to 96% RH, the devices were characterized both in a vacuum chamber and in a humidity chamber at atmospheric pressure. The measured response and recovery times of the graphene humidity sensors are on the order of several hundred milliseconds. Density functional theory simulations are employed to further investigate the sensitivity of the graphene devices towards water vapor. The interaction between the electrostatic dipole moment of the water and the impurity bands in the SiO2 substrate leads to electrostatic doping of the graphene layer. The proposed graphene sensor provides rapid response direct electrical readout and is compatible with back end of the line (BEOL) integration on top of CMOS-based integrated circuits.We demonstrate humidity sensing using a change of the electrical resistance of single-layer chemical vapor deposited (CVD) graphene that is placed on top of a SiO2 layer on a Si wafer. To investigate the selectivity of the sensor towards the most common constituents in air, its signal response was characterized individually for water vapor (H2O), nitrogen (N2), oxygen (O2), and argon (Ar). In order to assess the humidity sensing effect for a range from 1% relative humidity (RH) to 96% RH, the devices were characterized both in a vacuum chamber and in a humidity chamber at atmospheric pressure. The measured response and recovery times of the graphene humidity sensors are on the order of several hundred milliseconds. Density functional theory simulations are employed to further

  8. Direct Electrical Stimulation in the Human Brain Disrupts Melody Processing.

    Science.gov (United States)

    Garcea, Frank E; Chernoff, Benjamin L; Diamond, Bram; Lewis, Wesley; Sims, Maxwell H; Tomlinson, Samuel B; Teghipco, Alexander; Belkhir, Raouf; Gannon, Sarah B; Erickson, Steve; Smith, Susan O; Stone, Jonathan; Liu, Lynn; Tollefson, Trenton; Langfitt, John; Marvin, Elizabeth; Pilcher, Webster H; Mahon, Bradford Z

    2017-09-11

    Prior research using functional magnetic resonance imaging (fMRI) [1-4] and behavioral studies of patients with acquired or congenital amusia [5-8] suggest that the right posterior superior temporal gyrus (STG) in the human brain is specialized for aspects of music processing (for review, see [9-12]). Intracranial electrical brain stimulation in awake neurosurgery patients is a powerful means to determine the computations supported by specific brain regions and networks [13-21] because it provides reversible causal evidence with high spatial resolution (for review, see [22, 23]). Prior intracranial stimulation or cortical cooling studies have investigated musical abilities related to reading music scores [13, 14] and singing familiar songs [24, 25]. However, individuals with amusia (congenitally, or from a brain injury) have difficulty humming melodies but can be spared for singing familiar songs with familiar lyrics [26]. Here we report a detailed study of a musician with a low-grade tumor in the right temporal lobe. Functional MRI was used pre-operatively to localize music processing to the right STG, and the patient subsequently underwent awake intraoperative mapping using direct electrical stimulation during a melody repetition task. Stimulation of the right STG induced "music arrest" and errors in pitch but did not affect language processing. These findings provide causal evidence for the functional segregation of music and language processing in the human brain and confirm a specific role of the right STG in melody processing. VIDEO ABSTRACT. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. The green electricity market model. Proposal for an optional, cost-neutral direct marketing model for supplying electricity customers

    International Nuclear Information System (INIS)

    Heinemann, Ronald

    2014-01-01

    One of the main goals of the Renewable Energy Law (EEG) is the market integration of renewable energy resources. For this purpose it has introduced compulsory direct marketing on the basis of a moving market premium. At the same time the green electricity privilege, a regulation which made it possible for customers to be supplied with electricity from EEG plants, has been abolished without substitution with effect from 1 August 2014. This means that, aside from other direct marketing channels, which will not be economically viable save for in a few exceptional cases, it will no longer be possible in future to sell electricity from EEG plants to electricity customers under the designation ''electricity from renewable energy''. The reason for this is that electricity sold under the market premium model can no longer justifiably be said to originate from renewable energy. As a consequence, almost all green electricity products sold in Germany carry a foreign green electricity certificate.

  10. Active colloids as mobile microelectrodes for unified label-free selective cargo transport.

    Science.gov (United States)

    Boymelgreen, Alicia M; Balli, Tov; Miloh, Touvia; Yossifon, Gilad

    2018-02-22

    Utilization of active colloids to transport both biological and inorganic cargo has been widely examined in the context of applications ranging from targeted drug delivery to sample analysis. In general, carriers are customized to load one specific target via a mechanism distinct from that driving the transport. Here we unify these tasks and extend loading capabilities to include on-demand selection of multiple nano/micro-sized targets without the need for pre-labelling or surface functionalization. An externally applied electric field is singularly used to drive the active cargo carrier and transform it into a mobile floating electrode that can attract (trap) or repel specific targets from its surface by dielectrophoresis, enabling dynamic control of target selection, loading and rate of transport via the electric field parameters. In the future, dynamic selectivity could be combined with directed motion to develop building blocks for bottom-up fabrication in applications such as additive manufacturing and soft robotics.

  11. 78 FR 47235 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-08-05

    ...We propose to adopt a new airworthiness directive (AD) for certain General Electric Company (GE) model GEnx-2B67 and GEnx-2B67B turbofan engines. This proposed AD was prompted by the original equipment manufacturer's disclosure that certain critical rotating life-limited parts (LLPs) used in Boeing 747-8 flight tests had consumed more cyclic life than they would have in revenue flight cycles. These parts were then installed into engines and introduced into revenue service without adjustment to remaining cyclic life. This proposed AD would require a one-time adjustment to the cycle counts of those LLPs to account for the additional low cycle fatigue (LCF) life consumed during flight tests. We are proposing this AD to prevent the failure of critical rotating LLPs, uncontained engine failure, and damage to the airplane.

  12. Possibility of direct electricity production from waste canola oil

    Science.gov (United States)

    Włodarczyk, Paweł P.; Włodarczyk, Barbara; Kalinichenko, Antonina

    2017-10-01

    Powering high-efficiency devices, such as fuel cells, with waste products will allow for a broader development of renewable energy sources and utilisation of by- products. This publication presents the possibility of electrooxidation of the emulsion of waste rapeseed oil, prepared on the basis of the detergent Syntanol DS-10. The process of electrooxidation was carried out on platinum electrode in alkaline (KOH) and acidic (H2SO4) electrolyte, in the temperature range of 293-333 K. In each analysed case the process of electrooxidation took place. The maximum current density obtained was 7 mA cm-2. Thus, it has been shown that it is possible to generate electricity directly from the emulsion of the waste rapeseed oil.

  13. Possibility of direct electricity production from waste canola oil

    Directory of Open Access Journals (Sweden)

    Włodarczyk Paweł P.

    2017-01-01

    Full Text Available Powering high-efficiency devices, such as fuel cells, with waste products will allow for a broader development of renewable energy sources and utilisation of by- products. This publication presents the possibility of electrooxidation of the emulsion of waste rapeseed oil, prepared on the basis of the detergent Syntanol DS-10. The process of electrooxidation was carried out on platinum electrode in alkaline (KOH and acidic (H2SO4 electrolyte, in the temperature range of 293-333 K. In each analysed case the process of electrooxidation took place. The maximum current density obtained was 7 mA cm-2. Thus, it has been shown that it is possible to generate electricity directly from the emulsion of the waste rapeseed oil.

  14. The beauty of being (label)-free: sample preparation methods for SWATH-MS and next-generation targeted proteomics

    Science.gov (United States)

    Campbell, Kate; Deery, Michael J.; Lilley, Kathryn S.; Ralser, Markus

    2014-01-01

    The combination of qualitative analysis with label-free quantification has greatly facilitated the throughput and flexibility of novel proteomic techniques. However, such methods rely heavily on robust and reproducible sample preparation procedures. Here, we benchmark a selection of in gel, on filter, and in solution digestion workflows for their application in label-free proteomics. Each procedure was associated with differing advantages and disadvantages. The in gel methods interrogated were cost effective, but were limited in throughput and digest efficiency. Filter-aided sample preparations facilitated reasonable processing times and yielded a balanced representation of membrane proteins, but led to a high signal variation in quantification experiments. Two in solution digest protocols, however, gave optimal performance for label-free proteomics. A protocol based on the detergent RapiGest led to the highest number of detected proteins at second-best signal stability, while a protocol based on acetonitrile-digestion, RapidACN, scored best in throughput and signal stability but came second in protein identification. In addition, we compared label-free data dependent (DDA) and data independent (SWATH) acquisition on a TripleTOF 5600 instrument. While largely similar in protein detection, SWATH outperformed DDA in quantification, reducing signal variation and markedly increasing the number of precisely quantified peptides. PMID:24741437

  15. Data processing pipelines for comprehensive profiling of proteomics samples by label-free LC MS for biomarker discovery

    NARCIS (Netherlands)

    Christin, Christin; Bischoff, Rainer; Horvatovich, Peter

    2011-01-01

    Label-free quantitative LC-MS profiling of complex body fluids has become an important analytical tool for biomarker and biological knowledge discovery in the past decade. Accurate processing, statistical analysis and validation of acquired data diversified by the different types of mass

  16. Applying label-free dynamic mass redistribution assay for studying endogenous FPR1 receptor signalling in human neutrophils

    DEFF Research Database (Denmark)

    Christensen, Hanna B; Gloriam, David E; Pedersen, Daniel Sejer

    2017-01-01

    potencies. RESULTS: The potencies (pEC50) of fMLF, WKYMVm and compound 43, determined on human neutrophils using the label-free DMR assay were 8.63, 7.76 and 5.92, respectively. The DMR response to fMLF, but not WKYMVm and compound 43 could be blocked by the FPR1-specific antagonist cyclosporin H...

  17. Suppression of resonance Raman scattering via ground state depletion towards sub-diffraction-limited label-free microscopy

    NARCIS (Netherlands)

    Rieger, S.; Fischedick, M.; Boller, Klaus J.; Fallnich, Carsten

    2016-01-01

    We report on the first experimental demonstration of the suppression of spontaneous Raman scattering via ground state depletion. The concept of Raman suppression can be used to achieve sub-diffraction-limited resolution in label-free microscopy by exploiting spatially selective signal suppression

  18. Universal chitosan-assisted synthesis of Ag-including heterostructured nanocrystals for label-free in situ SERS monitoring.

    Science.gov (United States)

    Cai, Kai; Xiao, Xiaoyan; Zhang, Huan; Lu, Zhicheng; Liu, Jiawei; Li, Qin; Liu, Chen; Foda, Mohamed F; Han, Heyou

    2015-12-07

    A universal chitosan-assisted method was developed to synthesize various Ag-including heterostructured nanocrystals, in which chelation probably plays a vital role. The as-prepared Ag/Pd heterostructured nanocrystals show outstanding properties when used as bifunctional nanocomposites in label-free in situ SERS monitoring of Pd-catalyzed reaction.

  19. Selective label-free electrochemical impedance measurement of glycated haemoglobin on 3-aminophenylboronic acid-modified eggshell membranes

    DEFF Research Database (Denmark)

    Boonyasit, Yuwadee; Heiskanen, Arto; Chailapakul, Orawan

    2015-01-01

    We propose a novel alternative approach to long-term glycaemic monitoring using eggshell membranes (ESMs) as a new immobilising platform for the selective label-free electrochemical sensing of glycated haemoglobin (HbA1c), a vital clinical index of the glycaemic status in diabetic individuals. Due...

  20. DNA-hosted Hoechst dyes: application for label-free fluorescent monitoring of endonuclease activity and inhibition.

    Science.gov (United States)

    Jiang, Xiao-Qin; Guo, Su-Miao; Zhang, Min; Zhou, Ming; Ye, Bang-Ce

    2014-11-21

    A simple and facile approach was developed for monitoring EcoRI endonuclease activity and inhibition, in which a hairpin-like DNA containing restriction cutting site for EcoRI endonuclease acts as the sensing element and Hoechst dyes as the signal indicator in a label-free format.

  1. Electrokinetic label-free screening chip: a marriage of multiplexing and high throughput analysis using surface plasmon resonance imaging

    NARCIS (Netherlands)

    Krishnamoorthy, G.; Carlen, Edwin; Bomer, Johan G.; Wijnperle, Daniël; de Boer, Hans L.; van den Berg, Albert; Schasfoort, Richardus B.M.

    2010-01-01

    We present an electrokinetic label-free biomolecular screening chip (Glass/PDMS) to screen up to 10 samples simultaneously using surface plasmon resonance imaging (iSPR). This approach reduces the duration of an experiment when compared to conventional experimental methods. This new device offers a

  2. The goat (Capra hircus) mammary gland secretory tissue proteome as influenced by weight loss: A study using label free proteomics

    DEFF Research Database (Denmark)

    Hernandez Castellano, Lorenzo E; Ferreira, Ana M.; Nanni, Paolo

    2016-01-01

    Seasonal weight loss (SWL) is a significant limitation to animal production. Breeds that have evolved in harsh climates have acquired tolerance to SWL through selection. Herein, label free proteomics was used to characterize the effects of SWL in two goat breeds with different levels of adaptatio...

  3. Benchmarking quantitative label-free LC-MS data processing workflows using a complex spiked proteomic standard dataset.

    Science.gov (United States)

    Ramus, Claire; Hovasse, Agnès; Marcellin, Marlène; Hesse, Anne-Marie; Mouton-Barbosa, Emmanuelle; Bouyssié, David; Vaca, Sebastian; Carapito, Christine; Chaoui, Karima; Bruley, Christophe; Garin, Jérôme; Cianférani, Sarah; Ferro, Myriam; Van Dorssaeler, Alain; Burlet-Schiltz, Odile; Schaeffer, Christine; Couté, Yohann; Gonzalez de Peredo, Anne

    2016-01-30

    Proteomic workflows based on nanoLC-MS/MS data-dependent-acquisition analysis have progressed tremendously in recent years. High-resolution and fast sequencing instruments have enabled the use of label-free quantitative methods, based either on spectral counting or on MS signal analysis, which appear as an attractive way to analyze differential protein expression in complex biological samples. However, the computational processing of the data for label-free quantification still remains a challenge. Here, we used a proteomic standard composed of an equimolar mixture of 48 human proteins (Sigma UPS1) spiked at different concentrations into a background of yeast cell lysate to benchmark several label-free quantitative workflows, involving different software packages developed in recent years. This experimental design allowed to finely assess their performances in terms of sensitivity and false discovery rate, by measuring the number of true and false-positive (respectively UPS1 or yeast background proteins found as differential). The spiked standard dataset has been deposited to the ProteomeXchange repository with the identifier PXD001819 and can be used to benchmark other label-free workflows, adjust software parameter settings, improve algorithms for extraction of the quantitative metrics from raw MS data, or evaluate downstream statistical methods. Bioinformatic pipelines for label-free quantitative analysis must be objectively evaluated in their ability to detect variant proteins with good sensitivity and low false discovery rate in large-scale proteomic studies. This can be done through the use of complex spiked samples, for which the "ground truth" of variant proteins is known, allowing a statistical evaluation of the performances of the data processing workflow. We provide here such a controlled standard dataset and used it to evaluate the performances of several label-free bioinformatics tools (including MaxQuant, Skyline, MFPaQ, IRMa-hEIDI and Scaffold) in

  4. Supported binary liposome vesicle-gold nanoparticle for enhanced label free DNA and protein sensing.

    Science.gov (United States)

    Divya, Karutha Pandian; Dharuman, Venkataraman

    2017-09-15

    Supported binary liposome mixture of cationic liposome N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium propane (DOTAP) and the zwitterionic liposome 1,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine (DOPE) were tethered on thiol monolayers in the absence and presence of gold nanoparticle to enhance sensor stability and sensitivity for label free DNA and protein sensing for the first time. Cysteamine hydrochloride (Cyst), 3-Mercaptopropionic acid (MPA), 11-Mercaptoundecanoic acid (MUDA) and 11-amino-1-undecane thiol (AUT) monolayers were used as tethers on gold surfaces. Electrochemical studies in the presence of [Fe(CN) 6 ] 3-/4- indicate that the presence of both DOPE and AuNP decreases the electrostatic interaction between DOTAP and MPA layer during the formation of DOPE-DOTAP-AuNP (DDA) whereas they enhance the repulsive force on the Cyst and AUT monolayers. In the thiol monolayer supported DDA, the gelation of neutral lipid DOPE by the AuNP is disfavored which inturn promotes stability of vesicle structure. The membrane protein melittin's interaction with the DDA indicates the presence of intact vesicle by showing decreased charge transfer for the MUDA and AUT in the presence of [Fe(CN) 6 ] 3-/4- . On the contrary, the presence of the bilayer and semi circled DDA on the MPA and cysteamine layers were confirmed by the increased redox reaction. Atomic Force Microscopic (AFM) and Transmission Electron Microscopic (TEM) images support the presence of an array like semi circled DDA on the MPA and well separated DDA vesicles on the MUDA with variable sizes. Dynamic Light Scattering (DLS) and Fourier Transform Infrared spectroscopy (FTIR) suggest effective coordination between DOPE, DOTAP and AuNP. Label free DNA hybridization sensing in presence of the negatively charged [Fe(CN) 6 ] 3-/4- indicates the lowest DNA detection limit of 1×10 -14 M with linearity range 1×10 -13 to 1×10 -9 M. Similarly, streptavidin sensing shows the lowest detection of 1ngml -1 with a

  5. A regenerative label-free fiber optic sensor using surface plasmon resonance for clinical diagnosis of fibrinogen

    Directory of Open Access Journals (Sweden)

    Nguyen TT

    2015-08-01

    Full Text Available Tan Tai Nguyen,1 Sun Oh Bea,1 Dong Min Kim,2 Won Jung Yoon,3 Jin-Won Park,4 Seong Soo A An,1 Heongkyu Ju1,5,6 1Department of Bionano Technology, College of Bionano Technology, Gachon University, Seongnam, 2Department of Materials Science and Engineering, Hongik University, Sejong City, 3Department of Chemical and Bio Engineering, Gachon University, Seongnam, 4Department of Chemical and Biomolecular Engineering, College of Energy and Biotechnology, Seoul National University of Science and Technology, Seoul, 5Department of Nanophysics, College of Bionano Technology, Gachon University, Seongnam, 6Neuroscience Institute, Gil Hospital, Incheon, South Korea Purpose: We present the regenerative label-free fiber optical biosensor that exploits surface plasmon resonance for quantitative detection of fibrinogen (Fbg extracted from human blood plasma.Materials and methods: The sensor head was made up of a multimode optical fiber with its polymer cladding replaced by metal composite of nanometer thickness made of silver, aluminum, and nickel. The Ni layer coated allowed a direct immobilization of histidine-tagged peptide (HP on its metal surface without an additional cross-linker in between. On the coated HP layer, immunoglobulin G was then immobilized for specific capturing of Fbg.Results: We demonstrated a real-time quantitative detection of Fbg concentrations with limit of detection of ~10 ng/mL. The fact that the HP layer could be removed by imidazole with acid also permitted us to demonstrate the regeneration of the outermost metal surface of the sensor head for the sensor reusability.Conclusion: The sensor detection limit was estimated to be ~10 pM, which was believed to be sensitive enough for detecting Fbg during the clinical diagnosis of cardiovascular diseases, myocardial infarction, strokes, and Alzheimer’s diseases. Keywords: SPR, real-time assay, histidine-tagged peptide, protein sensing

  6. A label-free electrochemical immunosensor based on the novel signal amplification system of AuPdCu ternary nanoparticles functionalized polymer nanospheres.

    Science.gov (United States)

    Yan, Qin; Yang, Yuying; Tan, Zhaoling; Liu, Qing; Liu, Hui; Wang, Ping; Chen, Lei; Zhang, Daopeng; Li, Yueyun; Dong, Yunhui

    2018-04-30

    A sensitive label-free electrochemical immunosensor was designed using a novel signal amplification system for quantitative detecting hepatitis B surface antigen (HBsAg). Nitrogen-doped graphene quantum dots (N-GQDs) supported surfactant-free AuPdCu ternary nanoparticles (AuPdCu/N-GQDs), which featured with good conductivity and excellent catalytic properties for the reduction of hydrogen peroxide (H 2 O 2 ), was synthesized by a simple and benign hydrothermal procedure. At the same time, the electroactive polymer nanospheres (PS) was synthesized by infinite coordination polymers of ferrocenedicarboxylic acid, which could play as carrier and electronic mediator to load AuPdCu/N-GQDs. The PS not only improved the ability to load antibodies because of the good biocompatibility, but also accelerated electron transport of the electrode interface attribute to plentiful ferrocene unit. Thus, the prepared AuPdCu/N-GQDs@PS has abilities of good biocompatibility, catalytic activity and electrical conductivity to be applied as transducing materials to amplify electrochemical signal in detection of HBsAg. Under optimal conditions, the fabricated immunosensor exhibited high sensitivity and stability in the detection of HBsAg. A linear relationship between current signals and the concentrations of HBsAg was obtained in the range from 10fg/mL to 50ng/mL and the detection limit of HBsAg was 3.3fg/mL (signal-to-noise ratio of 3). Moreover, the designed immunosensor with excellent selectivity, reproducibility and stability shows excellent performance in detection of human serum samples. Furthermore, this label-free electrochemical immunosensor has promising application in clinical diagnosis of HBsAg. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Changing the direction and orientation of electric field during electric pulses application improves plasmid gene transfer in vitro.

    Science.gov (United States)

    Pavlin, Mojca; Haberl, Sasa A; Rebersek, Matej; Miklavcic, Damijan; Kanduser, Masa

    2011-09-12

    Gene electrotransfer is a physical method used to deliver genes into the cells by application of short and intense electric pulses, which cause destabilization of cell membrane, making it permeable to small molecules and allows transfer of large molecules such as DNA. It represents an alternative to viral vectors, due to its safety, efficacy and ease of application. For gene electrotransfer different electric pulse protocols are used in order to achieve maximum gene transfection, one of them is changing the electric field direction and orientation during the pulse delivery. Changing electric field direction and orientation increase the membrane area competent for DNA entry into the cell. In this video, we demonstrate the difference in gene electrotransfer efficacy when all pulses are delivered in the same direction and when pulses are delivered by changing alternatively the electric field direction and orientation. For this purpose tip with integrated electrodes and high-voltage prototype generator, which allows changing of electric field in different directions during electric pulse application, were used. Gene electrotransfer efficacy is determined 24h after pulse application as the number of cells expressing green fluorescent protein divided with the number of all cells. The results show that gene transfection is increased when the electric field orientation during electric pulse delivery is changed.

  8. Universal label-free in-process quantification of influenza virus-like particles.

    Science.gov (United States)

    Carvalho, Sofia B; Moleirinho, Mafalda G; Wheatley, David; Welsh, John; Gantier, René; Alves, Paula M; Peixoto, Cristina; Carrondo, Manuel J T

    2017-08-01

    Virus-like particles (VLPs) are becoming established as vaccines, in particular for influenza pandemics, increasing the interest in the development of VLPs manufacturing bioprocess. However, for complex VLPs, the analytical tools used for quantification are not yet able to keep up with the bioprocess progress. Currently, quantification for Influenza relies on traditional methods: hemagglutination assay or Single Radial Immunodiffusion. These analytical technologies are time-consuming, cumbersome, and not supportive of efficient downstream process development and monitoring. Hereby we report a label-free tool that uses Biolayer interferometry (BLI) technology applied on an Octet platform to quantify Influenza VLPs at all stages of bioprocess. Human (α2,6-linked sialic acid) and avian (α2,3-linked sialic acid) biotinylated receptors associated with streptavidin biosensors were used, to quantify hemagglutinin content in several mono- and multivalent Influenza VLPs. The applied method was able to quantify hemagglutinin from crude samples up to final bioprocessing VLP product. BLI technology confirmed its value as a high throughput analytical tool with high sensitivity and improved detection limits compared to traditional methods. This simple and fast method allowed for real-time results, which are crucial for in-line monitoring of downstream processing, improving process development, control and optimization. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Label-free signal-on aptasensor for sensitive electrochemical detection of arsenite.

    Science.gov (United States)

    Cui, Lin; Wu, Jie; Ju, Huangxian

    2016-05-15

    A signal-on aptasensor was fabricated for highly sensitive and selective electrochemical detection of arsenite with a label-free Ars-3 aptamer self-assembled on a screen-printed carbon electrode (SPCE) via Au-S bond. The Ars-3 aptamer could adsorb cationic polydiallyldimethylammonium (PDDA) via electrostatic interaction to repel other cationic species. In the presence of arsenite, the change of Ars-3 conformation due to the formation of Ars-3/arsenite complex led to less adsorption of PDDA, and the complex could adsorb more positively charged [Ru(NH3)6](3+) as an electrochemically active indicator on the aptasensor surface, which produced a sensitive "turn-on" response. The target-induced structure switching could be used for sensitive detection of arsenite with a linear range from 0.2 nM to 100 nM and a detection limit down to 0.15 nM. Benefiting from Ars-3 aptamer, the proposed system exhibited excellent specificity against other heavy metal ions. The SPCE-based aptasensor exhibited the advantages of low cost and simple fabrication, providing potential application of arsenite detection in environment. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Label-free detection of interleukin-6 using electrolyte gated organic field effect transistors.

    Science.gov (United States)

    Diacci, Chiara; Berto, Marcello; Di Lauro, Michele; Bianchini, Elena; Pinti, Marcello; Simon, Daniel T; Biscarini, Fabio; Bortolotti, Carlo A

    2017-09-27

    Cytokines are small proteins that play fundamental roles in inflammatory processes in the human body. In particular, interleukin (IL)-6 is a multifunctional cytokine, whose increased levels are associated with infection, cancer, and inflammation. The quantification of IL-6 is therefore of primary importance in early stages of inflammation and in chronic diseases, but standard techniques are expensive, time-consuming, and usually rely on fluorescent or radioactive labels. Organic electronic devices and, in particular, organic field-effect transistors (OFETs) have been proposed in the recent years as novel platforms for label-free protein detection, exploiting as sensing unit surface-immobilized antibodies or aptamers. Here, the authors report two electrolyte-gated OFETs biosensors for IL-6 detection, featuring monoclonal antibodies and peptide aptamers adsorbed at the gate. Both strategies yield biosensors that can work on a wide range of IL-6 concentrations and exhibit a remarkable limit of detection of 1 pM. Eventually, electrolyte gated OFETs responses have been used to extract and compare the binding thermodynamics between the sensing moiety, immobilized at the gate electrode, and IL-6.

  11. Mueller matrix microscopy for label-free histopathology examinations (Conference Presentation)

    Science.gov (United States)

    Lv, Donghong; He, Honghui; Zhou, Jialing; Ma, Hui

    2017-03-01

    In clinical medicine, a pathologist often needs to examine cells or thin slices of tissues to identify abnormalities that are markers or precursors of diseases. Various chemical and immunohistochemical staining techniques have been developed to selectively label certain components to bring up the contrasts of specific microstructures. It is well known that a Mueller matrix contains rich information on the microstructure and optical properties of a sample. Using proper data analysis techniques, Mueller matrix images can also be transformed into new polarization parameters sensitive only to specific microstructural features. These new polarization parameters can selectively enhance the contrast of specific features in images of unstained pathological slide to help identify abnormalities. In recent studies, we set up a modulus design Mueller matrix microscope by adding polarization optics components into the optical path of a commercial transmission microscope. We take multiple measurements of the unstained pathological slide at different polar and azimuth angles, then derive an intrinsic Mueller matrix (IMM) which represents only the microstructural characters of the sample without the interference by the sample orientation. Such orientation-independent IMM images preserve to the maximum extent the pathological information of the tissue samples. Using Mueller matrix decomposition and transformation techniques, we demonstrate in preliminary tests that we are able to selectively enhance different characteristic features in different cancer tissues. With the fast advances in big-data analysis techniques, it is expected that label-free Mueller matrix microscopy is a potentially powerful tool for the histopathologists to identify characteristic features in complex tissue samples.

  12. Label-free multidimensional information acquisition from optogenetically engineered cells using a graphene transistor.

    Science.gov (United States)

    Li, Gongxin; Yang, Jia; Yang, Wenguang; Wang, Feifei; Wang, Yuechao; Wang, Wenxue; Liu, Lianqing

    2018-02-01

    The optogenetic technique, which allows the manipulation of cellular activity patterns in space and time by light, has transformed the field of neuroscience. However, acquiring multidimensional optogenetic information remains challenging despite the fact that several cellular information detection methods have been proposed. Herein, we present a new method to acquire label-free multidimensional information from optogenetically engineered cells using a graphene transistor. Using a graphene film to form a strong densely packed layer with cells, the cellular action potentials were characterized as light-activated transistor conductance signals, which quantified the multidimensional optogenetic information. Based on this approach, some important cellular optogenetic information, including electrophysiological state, cell concentration, expression levels of opsin and response to variable light intensity, were also precisely detected. Furthermore, the graphene transistor was also used to distinguish cells expressing different channelrhodopsin-2 variants. Our study offers a general detection method of multidimensional optogenetic information for extending the applications of the optogenetic technique and provides a novel sensor for the development of future biological prosthetic devices.

  13. Minimizing technical variation during sample preparation prior to label-free quantitative mass spectrometry.

    Science.gov (United States)

    Scheerlinck, E; Dhaenens, M; Van Soom, A; Peelman, L; De Sutter, P; Van Steendam, K; Deforce, D

    2015-12-01

    Sample preparation is the crucial starting point to obtain high-quality mass spectrometry data and can be divided into two main steps in a bottom-up proteomics approach: cell/tissue lysis with or without detergents and a(n) (in-solution) digest comprising denaturation, reduction, alkylation, and digesting of the proteins. Here, some important considerations, among others, are that the reagents used for sample preparation can inhibit the digestion enzyme (e.g., 0.1% sodium dodecyl sulfate [SDS] and 0.5 M guanidine HCl), give rise to ion suppression (e.g., polyethylene glycol [PEG]), be incompatible with liquid chromatography-tandem mass spectrometry (LC-MS/MS) (e.g., SDS), and can induce additional modifications (e.g., urea). Taken together, all of these irreproducible effects are gradually becoming a problem when label-free quantitation of the samples is envisioned such as during the increasingly popular high-definition mass spectrometry (HDMS(E)) and sequential window acquisition of all theoretical fragment ion spectra (SWATH) data-independent acquisition strategies. Here, we describe the detailed validation of a reproducible method with sufficient protein yield for sample preparation without any known LC-MS/MS interfering substances by using 1% sodium deoxycholate (SDC) during both cell lysis and in-solution digest. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  14. Label-Free Imaging of Nanoparticle Uptake Competition in Single Cells by Hyperspectral Stimulated Raman Scattering.

    Science.gov (United States)

    Huang, Bin; Yan, Shuai; Xiao, Lin; Ji, Rong; Yang, Liuyan; Miao, Ai-Jun; Wang, Ping

    2018-03-01

    Imaging and quantification of nanoparticles in single cells in their most natural condition are expected to facilitate the biotechnological applications of nanoparticles and allow for better assessment of their biosafety risks. However, current imaging modalities either require tedious sample preparation or only apply to nanoparticles with specific physicochemical characteristics. Here, the emerging hyperspectral stimulated Raman scattering (SRS) microscopy, as a label-free and nondestructive imaging method, is used for the first time to investigate the subcellular distribution of nanoparticles in the protozoan Tetrahymena thermophila. The two frequently studied nanoparticles, polyacrylate-coated α-Fe 2 O 3 and TiO 2 , are found to have different subcellular distribution pattern as a result of their dissimilar uptake routes. Significant uptake competition between these two types of nanoparticles is further discovered, which should be paid attention to in future bioapplications of nanoparticles. Overall, this study illustrates the great promise of hyperspectral SRS as an analytical imaging tool in nanobiotechnology and nanotoxicology. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Label-Free Raman Hyperspectral Imaging of Single Cells Cultured on Polymer Substrates.

    Science.gov (United States)

    Sinjab, Faris; Sicilia, Giovanna; Shipp, Dustin W; Marlow, Maria; Notingher, Ioan

    2017-12-01

    While Raman hyperspectral imaging has been widely used for label-free mapping of biomolecules in cells, these measurements require the cells to be cultured on weakly Raman scattering substrates. However, many applications in biological sciences and engineering require the cells to be cultured on polymer substrates that often generate large Raman scattering signals. Here, we discuss the theoretical limits of the signal-to-noise ratio in the Raman spectra of cells in the presence of polymer signals and how optical aberrations may affect these measurements. We show that Raman spectra of cells cultured on polymer substrates can be obtained using automatic subtraction of the polymer signals and demonstrate the capabilities of these methods in two important applications: tissue engineering and in vitro toxicology screening of drugs. Apart from their scientific and technological importance, these applications are examples of the two most common measurement configurations: (1) cells cultured on an optically thick polymer substrate measured using an immersion/dipping objective; and (2) cells cultured on a transparent polymer substrate and measured using an inverted optical microscope. In these examples, we show that Raman hyperspectral data sets with sufficient quality can be successfully acquired to map the distribution of common biomolecules in cells, such as nucleic acids, proteins, and lipids, as well as detecting the early stages of apoptosis. We also discuss strategies for further improvements that could expand the application of Raman hyperspectral imaging on polymer substrates even further in biomedical sciences and engineering.

  16. Label-free haemogram using wavelength modulated Raman spectroscopy for identifying immune-cell subset

    Science.gov (United States)

    Ashok, Praveen C.; Praveen, Bavishna B.; Campbell, Elaine C.; Dholakia, Kishan; Powis, Simon J.

    2014-03-01

    Leucocytes in the blood of mammals form a powerful protective system against a wide range of dangerous pathogens. There are several types of immune cells that has specific role in the whole immune system. The number and type of immune cells alter in the disease state and identifying the type of immune cell provides information about a person's state of health. There are several immune cell subsets that are essentially morphologically identical and require external labeling to enable discrimination. Here we demonstrate the feasibility of using Wavelength Modulated Raman Spectroscopy (WMRS) with suitable machine learning algorithms as a label-free method to distinguish between different closely lying immune cell subset. Principal Component Analysis (PCA) was performed on WMRS data from single cells, obtained using confocal Raman microscopy for feature reduction, followed by Support Vector Machine (SVM) for binary discrimination of various cell subset, which yielded an accuracy >85%. The method was successful in discriminating between untouched and unfixed purified populations of CD4+CD3+ and CD8+CD3+ T lymphocyte subsets, and CD56+CD3- natural killer cells with a high degree of specificity. It was also proved sensitive enough to identify unique Raman signatures that allow clear discrimination between dendritic cell subsets, comprising CD303+CD45+ plasmacytoid and CD1c+CD141+ myeloid dendritic cells. The results of this study clearly show that WMRS is highly sensitive and can distinguish between cell types that are morphologically identical.

  17. Label-free identification of white blood cell using optical diffraction tomography (Conference Presentation)

    Science.gov (United States)

    Yoon, Jonghee; Kim, Kyoohyun; Kim, Min-hyeok; Kang, Suk-Jo; Park, YongKeun

    2016-03-01

    White blood cells (WBC) have crucial roles in immune systems which defend the host against from disease conditions and harmful invaders. Various WBC subsets have been characterized and reported to be involved in many pathophysiologic conditions. It is crucial to isolate a specific WBC subset to study its pathophysiological roles in diseases. Identification methods for a specific WBC population are rely on invasive approaches, including Wright-Gimesa staining for observing cellular morphologies and fluorescence staining for specific protein markers. While these methods enable precise classification of WBC populations, they could disturb cellular viability or functions. In order to classify WBC populations in a non-invasive manner, we exploited optical diffraction tomography (ODT). ODT is a three-dimensional (3-D) quantitative phase imaging technique that measures 3-D refractive index (RI) distributions of individual WBCs. To test feasibility of label-free classification of WBC populations using ODT, we measured four subtypes of WBCs, including B cell, CD4 T cell, CD8 T cell, and natural killer (NK) cell. From measured 3-D RI tomograms of WBCs, we obtain quantitative structural and biochemical information and classify each WBC population using a machine learning algorithm.

  18. Label-free quantitative proteome analysis of the surface-bound salivary pellicle.

    Science.gov (United States)

    Delius, Judith; Trautmann, Simone; Médard, Guillaume; Kuster, Bernhard; Hannig, Matthias; Hofmann, Thomas

    2017-04-01

    The salivary pellicle, covering natural as well as restored tooth surfaces in the oral cavity as an immobilized protein-rich layer, acts as an important physico-chemical and biological mediator at the tooth-saliva-interface. For the first time, the pellicle's proteome of individual volunteers were analyzed separately on three consecutive days and the relative protein abundance determined by a label-free quantitative nano-LC-MS/MS approach. A total of 72 major proteins were identified in the initial pellicles formed intraorally on dental ceramic specimens already after 3min with high inter-individual and inter-day consistency. In comparison, significant differences in protein abundance were evident between subjects, thus indicating unique individual pellicle profiles. Furthermore, the relative protein abundance in pellicles was compared to the proteome pattern in the corresponding saliva samples of the same individuals to provide first data on significantly enriched and depleted salivary proteins (p <0.05) within the surface-bound salivary pellicle. Our findings reveal the initial adsorption of salivary proteins at the solid-liquid interface to be a rapid, highly selective, and reproducible process leading to the immobilization of a broad range of protective proteins and enzymes on the substratum surface within a few minutes. This provides evidence that the pellicle layer might be physiologically functional even without further maturation. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Design of a New Ultracompact Resonant Plasmonic Multi-Analyte Label-Free Biosensing Platform

    Directory of Open Access Journals (Sweden)

    Francesco Dell’Olio

    2017-08-01

    Full Text Available In this paper, we report on the design of a bio-multisensing platform for the selective label-free detection of protein biomarkers, carried out through a 3D numerical algorithm. The platform includes a number of biosensors, each of them is based on a plasmonic nanocavity, consisting of a periodic metal structure to be deposited on a silicon oxide substrate. Light is strongly confined in a region with extremely small size (=1.57 μm2, to enhance the light-matter interaction. A surface sensitivity Ss = 1.8 nm/nm has been calculated together with a detection limit of 128 pg/mm2. Such performance, together with the extremely small footprint, allow the integration of several devices on a single chip to realize extremely compact lab-on-chip microsystems. In addition, each sensing element of the platform has a good chemical stability that is guaranteed by the selection of gold for its fabrication.

  20. Tapered Optical Fiber Sensor for Label-Free Detection of Biomolecules

    Directory of Open Access Journals (Sweden)

    Xingwei Wang

    2011-03-01

    Full Text Available This paper presents a fast, highly sensitive and low-cost tapered optical fiber biosensor that enables the label-free detection of biomolecules. The sensor takes advantage of the interference effect between the fiber’s first two propagation modes along the taper waist region. The biomolecules bonded on the taper surface were determined by demodulating the transmission spectrum phase shift. Because of the sharp spectrum fringe signals, as well as a relatively long biomolecule testing region, the sensor displayed a fast response and was highly sensitive. To better understand the influence of various biomolecules on the sensor, a numerical simulation that varied biolayer parameters such as thickness and refractive index was performed. The results showed that the spectrum fringe shift was obvious to be measured even when the biolayer was only nanometers thick. A microchannel chip was designed and fabricated for the protection of the sensor and biotesting. Microelectromechanical systems (MEMS fabrication techniques were used to precisely control the profile and depth of the microchannel on the silicon chip with an accuracy of 2 μm. A tapered optical fiber biosensor was fabricated and evaluated with an Immune globulin G (IgG antibody-antigen pair.

  1. Luminescent Metal-Organic-Framework-Based Label-Free Assay of Polyphenol Oxidase with Fluorescent Scan.

    Science.gov (United States)

    Li, Yue; Guo, An; Chang, Lan; Li, Wen-Juan; Ruan, Wen-Juan

    2017-05-11

    Metal-organic frameworks (MOFs) are emerging in recent years as a kind of versatile fluorescent sensing materials, but their application to enzyme assays has rarely been studied. Here, the first example of a MOF-based label-free enzyme assay system is reported. A luminescent MOF was synthesized and applied to the activity analysis of polyphenol oxidase (PPO). With its distinct responses to the phenolic substrate and o-quinone product, this MOF could transduce the extent of PPO-catalyzed oxidation to fluorescence signal and enable the real-time monitoring of this reaction. Wide substrate adaptability and high sensitivity (detection limit=0.00012 U mL -1 ) were exhibited by this method, which meets the requirement of common bioanalysis. Interestingly, by the comparison with molecular capturing reagents, the heterogeneous nature of this MOF-based assay effectively preventing the interaction with the enzyme was proven, thus ensuring the authenticity of results. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. KF polymerase-based fluorescence aptasensor for the label-free adenosine detection.

    Science.gov (United States)

    Liao, Dongli; Jiao, Huping; Wang, Bin; Lin, Quan; Yu, Cong

    2012-02-21

    We have developed a simple, inexpensive, and label-free method for the selective detection of adenosine. Klenow fragment polymerase (KF polymerase) is a commonly-used 5' to 3' DNA polymerase, it also has 3' to 5' exonuclease activity that can digest single-stranded DNA. An adenosine binding DNA aptamer was employed, the aptamer was split into two pieces of single-stranded DNA (aptamer-A1 + aptamer-A2). Without the addition of adenosine, aptamer-A1 and aptamer-A2 existed as single-stranded DNA which could be efficiently degraded by the exonuclease activity of KF polymerase. Much reduced background fluorescence was obtained when SYBR Green dye was added. However, in the presence of adenosine, aptamer-A1 and aptamer-A2 bound to adenosine, and hybridization of the complementary sequences resulted in the formation of a duplex DNA structure, which could initiate DNA polymerization. The addition of SYBR Green dye resulted in a very high fluorescence enhancement, which could be used for the quantification of adenosine.

  3. A label free aptasensor for Ochratoxin A detection in cocoa beans: An application to chocolate industries.

    Science.gov (United States)

    Mishra, Rupesh K; Hayat, Akhtar; Catanante, Gaëlle; Ocaña, Cristina; Marty, Jean-Louis

    2015-08-19

    Contamination of food by mycotoxin occurs in minute/trace quantities. Nearly 92.5% of the cocoa samples present Ochratoxin A (OTA) levels at trace quantity. Hence, there is a necessity for a highly sensitive and selective device that can detect and quantify these organic toxins in various matrices such as cocoa beans. This work reports for the first time, a facile and label-free electrochemical impedimetric aptasensor for rapid detection and quantitation of OTA in cocoa beans. The developed aptasensor was constructed based on the diazonium-coupling reaction mechanism for the immobilization of anti-OTA-aptamer on screen printed carbon electrodes (SPCEs). The aptasensor exhibited a very good limit of detection (LOD) as low as 0.15 ng/mL, with added advantages of good selectivity and reproducibility. The increase in electron transfer resistance was linearly proportional to the OTA concentration in the range 0.15-2.5 ng/mL, with an acceptable recovery percentage (91-95%, RSD = 4.8%) obtained in cocoa samples. This work can facilitate a general model for the detection of OTA in cocoa beans based on the impedimetric aptasensor. The analysis can be performed onsite with pre-constructed and aptamer modified electrodes employing a portable EIS set up. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells

    Directory of Open Access Journals (Sweden)

    Raf Donders

    2016-01-01

    Full Text Available In situ detection of MSCs remains difficult and warrants additional methods to aid with their characterization in vivo. Two-photon confocal laser scanning microscopy (TPM and second harmonic generation (SHG could fill this gap. Both techniques enable the detection of cells and extracellular structures, based on intrinsic properties of the specific tissue and intracellular molecules under optical irradiation. TPM imaging and SHG imaging have been used for label-free monitoring of stem cells differentiation, assessment of their behavior in biocompatible scaffolds, and even cell tracking in vivo. In this study, we show that TPM and SHG can accurately depict the umbilical cord architecture and visualize individual cells both in situ and during culture initiation, without the use of exogenously applied labels. In combination with nuclear DNA staining, we observed a variance in fluorescent intensity in the vessel walls. In addition, antibody staining showed differences in Oct4, αSMA, vimentin, and ALDH1A1 expression in situ, indicating functional differences among the umbilical cord cell populations. In future research, marker-free imaging can be of great added value to the current antigen-based staining methods for describing tissue structures and for the identification of progenitor cells in their tissue of origin.

  5. A G-quadruplex-based Label-free Fluorometric Aptasensor for Adenosine Triphosphate Detection.

    Science.gov (United States)

    Li, Li Juan; Tian, Xue; Kong, Xiang Juan; Chu, Xia

    2015-01-01

    A G-quadruplex-based, label-free fluorescence assay was demonstrated for the detection of adenosine triphosphate (ATP). A double-stranded DNA (dsDNA), hybridized by ATP-aptamer and its complementary sequence, was employed as a substrate for ATP binding. SYBR Green I (SG I) was a fluorescent probe and exonuclease III (Exo III) was a nuclease to digest the dsDNA. Consequently, in the absence of ATP, the dsDNA was inset with SG I and was digested by Exo III, resulting in a low background signal. In the presence of ATP, the aptamer in dsDNA folded into a G-quadruplex structure that resisted the digestion of Exo III. SG I was inserted into the structure, showing high fluorescence. Owing to a decrease of the background noise, a high signal-to-noise ratio could be obtained. This sensor can detect ATP with a concentration ranging from 50 μM to 5 mM, and possesses a capacity for the sensitive determination of other targets.

  6. Label-free imaging of rat spinal cords based on multiphoton microscopy

    Science.gov (United States)

    Liao, Chenxi; Wang, Zhenyu; Zhou, Linquan; Zhu, Xiaoqin; Liu, Wenge; Chen, Jianxin

    2016-10-01

    As an integral part of the central nervous system, the spinal cord is a communication cable between the body and the brain. It mainly contains neurons, glial cells, nerve fibers and fiber tracts. The recent development of the optical imaging technique allows high-resolution imaging of biological tissues with the great potential for non-invasively looking inside the body. In this work, we evaluate the imaging capacity of multiphoton microscopy (MPM) based on second harmonic generation (SHG) and two-photon excited fluorescence (TPEF) for the cells and extracellular matrix in the spinal cord at molecular level. Rat spinal cord tissues were sectioned and imaged by MPM to demonstrate that MPM is able to show the microstructure including white matter, gray matter, ventral horns, dorsal horns, and axons based on the distinct intrinsic sources in each region of spinal cord. In the high-resolution and high-contrast MPM images, the cell profile can be clearly identified as dark shadows caused by nuclei and encircled by cytoplasm. The nerve fibers in white matter region emitted both SHG and TPEF signals. The multiphoton microscopic imaging technique proves to be a fast and effective tool for label-free imaging spinal cord tissues, based on endogenous signals in biological tissue. It has the potential to extend this optical technique to clinical study, where the rapid and damage-free imaging is needed.

  7. A distributed national network for label-free rapid identification of emerging pathogens

    Science.gov (United States)

    Robinson, J. Paul; Rajwa, Bartek P.; Dundar, M. Murat; Bae, Euiwon; Patsekin, Valery; Hirleman, E. Daniel; Roumani, Ali; Bhunia, Arun K.; Dietz, J. Eric; Davisson, V. Jo; Thomas, John G.

    2011-05-01

    Typical bioterrorism prevention scenarios assume well-known and well-characterized pathogens like anthrax or tularemia, which are serious public concerns if released into food and/or water supplies or distributed using other vectors. Common governmental contingencies include rapid response to these biological threats with predefined treatments and management operations. However, bioterrorist attacks may follow a far more sophisticated route. With the widely known and immense progress in genetics and the availability of molecular biology tools worldwide, the potential for malicious modification of pathogenic genomes is very high. Common non-pathogenic microorganisms could be transformed into dangerous, debilitating pathogens. Known pathogens could also be modified to avoid detection, because organisms are traditionally identified on the basis of their known physiological or genetic properties. In the absence of defined primers a laboratory using genetic biodetection methods such as PCR might be unable to quickly identify a modified microorganism. Our concept includes developing a nationwide database of signatures based on biophysical (such as elastic light scattering (ELS) properties and/or Raman spectra) rather than genetic properties of bacteria. When paired with a machine-learning system for emerging pathogen detection these data become an effective detection system. The approach emphasizes ease of implementation using a standardized collection of phenotypic information and extraction of biophysical features of pathogens. Owing to the label-free nature of the detection modalities ELS is significantly less costly than any genotypic or mass spectrometry approach.

  8. Study on a luminol-based electrochemiluminescent sensor for label-free DNA sensing.

    Science.gov (United States)

    Chu, Hai-Hong; Yan, Ji-Lin; Tu, Yi-Feng

    2010-01-01

    Automatic, inexpensive, simple and sensitive methods for DNA sensing and quantification are highly desirable for biomedical research. The rapid development of both the fundamentals and applications of electrochemiluminescence (ECL) over the past years has demonstrated its potential for analytical and bio-analytical chemistry. This paper reports the quenching effect of DNA on the ECL of luminol and the further development of a DNA sensing device. With the pre-functionalization by a composite of carbon nano-tubes (CNTs) and Au nanoparticles (AuNPs), the sensor provides a novel and valuable label-free approach for DNA sensing. Here the ECL intensity was remarkably decreased when more than 1.0 × 10(-12) molar of DNA were adsorbed on the sensor. Linearity of the DNA amount with the reciprocal of ECL intensity was observed. A saturated sensor caused a 92.8% quenching effect. The research also proposes the mechanism for the quenching effect which could be attributed to the interaction between luminol and DNA and the elimination of reactive oxygen species (ROSs) by DNA.

  9. Study on a Luminol-based Electrochemiluminescent Sensor for Label-Free DNA Sensing

    Directory of Open Access Journals (Sweden)

    Yi-Feng Tu

    2010-10-01

    Full Text Available Automatic, inexpensive, simple and sensitive methods for DNA sensing and quantification are highly desirable for biomedical research. The rapid development of both the fundamentals and applications of electrochemiluminescence (ECL over the past years has demonstrated its potential for analytical and bio-analytical chemistry. This paper reports the quenching effect of DNA on the ECL of luminol and the further development of a DNA sensing device. With the pre-functionalization by a composite of carbon nano-tubes (CNTs and Au nanoparticles (AuNPs, the sensor provides a novel and valuable label-free approach for DNA sensing. Here the ECL intensity was remarkably decreased when more than 1.0 × 10−12 molar of DNA were adsorbed on the sensor. Linearity of the DNA amount with the reciprocal of ECL intensity was observed. A saturated sensor caused a 92.8% quenching effect. The research also proposes the mechanism for the quenching effect which could be attributed to the interaction between luminol and DNA and the elimination of reactive oxygen species (ROSs by DNA.

  10. Label free quantitative proteomics analysis on the cisplatin resistance in ovarian cancer cells.

    Science.gov (United States)

    Wang, F; Zhu, Y; Fang, S; Li, S; Liu, S

    2017-05-20

    Quantitative proteomics has been made great progress in recent years. Label free quantitative proteomics analysis based on the mass spectrometry is widely used. Using this technique, we determined the differentially expressed proteins in the cisplatin-sensitive ovarian cancer cells COC1 and cisplatin-resistant cells COC1/DDP before and after the application of cisplatin. Using the GO analysis, we classified those proteins into different subgroups bases on their cellular component, biological process, and molecular function. We also used KEGG pathway analysis to determine the key signal pathways that those proteins were involved in. There are 710 differential proteins between COC1 and COC1/DDP cells, 783 between COC1 and COC1/DDP cells treated with cisplatin, 917 between the COC1/DDP cells and COC1/DDP cells treated with LaCl3, 775 between COC1/DDP cells treated with cisplatin and COC1/DDP cells treated with cisplatin and LaCl3. Among the same 411 differentially expressed proteins in cisplatin-sensitive COC1 cells and cisplain-resistant COC1/DDP cells before and after cisplatin treatment, 14% of them were localized on the cell membrane. According to the KEGG results, differentially expressed proteins were classified into 21 groups. The most abundant proteins were involved in spliceosome. This study lays a foundation for deciphering the mechanism for drug resistance in ovarian tumor.

  11. Label-free optical detection of bacteria on a 1-D photonic crystal of porous silicon

    Science.gov (United States)

    Wu, Chia-Chen; Alvarez, Sara D.; Rang, Camilla U.; Chao, Lin; Sailor, Michael J.

    2009-02-01

    The construction of a specific, label-free, bacteria biosensor using porous silicon 1-D photonic crystals will be described. Bacteria resident on the surface of porous silicon act as scattering centers for light resonant with the photonic crystal; the diffusely scattered light possesses the optical spectrum of the underlying photonic crystal. Using a spectrometer fitted to a light microscope, the bacteria are imaged without using exogenous dyes or labels and are quantified by measuring the intensity of scattered light. In order to selectively bind and identify bacteria using porous Si, we use surface modifications to reduce nonspecific binding to the surface and to engineer bacteria specificity onto the surface. Bovine serum albumin (BSA) was adsorbed to the porous Si surface to reduce nonspecific binding of bacteria. The coatings were then chemically activated to immobilize polyclonal antibodies specific to Escherichia coli. Two E. coli strains were used in our study, E. coli DH5α and non-pathogenic enterohemorrhagic Escherichia coli (EHEC) strain. The nonpathogenic Vibrio cholerae O1 strain was used to test for antibody specificity. Successful attachment of antibodies was measured using fluorescence microscopy and the scattering method was used to test for bacteria binding specificity.

  12. Classification of blood cells and tumor cells using label-free ultrasound and photoacoustics.

    Science.gov (United States)

    Strohm, Eric M; Kolios, Michael C

    2015-08-01

    A label-free method that can identify cells in a blood sample using high frequency photoacoustic and ultrasound signals is demonstrated. When the wavelength of the ultrasound or photoacoustic wave is similar to the size of a single cell (frequencies of 100-500 MHz), unique periodic features occur within the ultrasound and photoacoustic power spectrum that depend on the cell size, structure, and morphology. These spectral features can be used to identify different cell types present in blood, such as red blood cells (RBCs), white blood cells (WBCs), and circulating tumor cells. Circulating melanoma cells are ideal for photoacoustic detection due to their endogenous optical absorption properties. Using a 532 nm pulsed laser and a 375 MHz transducer, the ultrasound and photoacoustic signals from RBCs, WBCs, and melanoma cells were individually measured in an acoustic microscope to examine how the signals change between cell types. A photoacoustic and ultrasound signal was detected from RBCs and melanoma cells; only an ultrasound signal was detected from WBCs. The different cell types were distinctly separated using the ultrasound and photoacoustic signal amplitude and power spectral periodicity. The size of each cell was also estimated from the spectral periodicity. For the first time, sound waves generated using pulse-echo ultrasound and photoacoustics have been used to identify and size single cells, with applications toward counting and identifying cells, including circulating melanoma cells. © 2015 International Society for Advancement of Cytometry.

  13. Divergent Label-free Cell Phenotypic Pharmacology of Ligands at the Overexpressed β2-Adrenergic Receptors

    Science.gov (United States)

    Ferrie, Ann M.; Sun, Haiyan; Zaytseva, Natalya; Fang, Ye

    2014-01-01

    We present subclone sensitive cell phenotypic pharmacology of ligands at the β2-adrenergic receptor (β2-AR) stably expressed in HEK-293 cells. The parental cell line was transfected with green fluorescent protein (GFP)-tagged β2-AR. Four stable subclones were established and used to profile a library of sixty-nine AR ligands. Dynamic mass redistribution (DMR) profiling resulted in a pharmacological activity map suggesting that HEK293 endogenously expresses functional Gi-coupled α2-AR and Gs-coupled β2-AR, and the label-free cell phenotypic activity of AR ligands are subclone dependent. Pathway deconvolution revealed that the DMR of epinephrine is originated mostly from the remodeling of actin microfilaments and adhesion complexes, to less extent from the microtubule networks and receptor trafficking, and certain agonists displayed different efficacy towards the cAMP-Epac pathway. We demonstrate that receptor signaling and ligand pharmacology is sensitive to the receptor expression level, and the organization of the receptor and its signaling circuitry.

  14. Cell Surface Proteome of Dental Pulp Stem Cells Identified by Label-Free Mass Spectrometry.

    Directory of Open Access Journals (Sweden)

    Christian Niehage

    Full Text Available Multipotent mesenchymal stromal cells (MSCs are promising tools for regenerative medicine. They can be isolated from different sources based on their plastic-adherence property. The identification of reliable cell surface markers thus becomes the Holy Grail for their prospective isolation. Here, we determine the cell surface proteomes of human dental pulp-derived MSCs isolated from single donors after culture expansion in low (2% or high (10% serum-containing media. Cell surface proteins were tagged on intact cells using cell impermeable, cleavable sulfo-NHS-SS-biotin, which allows their enrichment by streptavidin pull-down. For the proteomic analyses, we first compared label-free methods to analyze cell surface proteomes i.e. composition, enrichment and proteomic differences, and we developed a new mathematical model to determine cell surface protein enrichment using a combinatorial gene ontology query. Using this workflow, we identified 101 cluster of differentiation (CD markers and 286 non-CD cell surface proteins. Based on this proteome profiling, we identified 14 cell surface proteins, which varied consistently in abundance when cells were cultured under low or high serum conditions. Collectively, our analytical methods provide a basis for identifying the cell surface proteome of dental pulp stem cells isolated from single donors and its evolution during culture or differentiation. Our data provide a comprehensive cell surface proteome for the precise identification of dental pulp-derived MSC populations and their isolation for potential therapeutic intervention.

  15. A Label-Free Photoluminescence Genosensor Using Nanostructured Magnesium Oxide for Cholera Detection

    Science.gov (United States)

    Patel, Manoj Kumar; Ali, Md. Azahar; Krishnan, Sadagopan; Agrawal, Ved Varun; Al Kheraif, Abdulaziz A.; Fouad, H.; Ansari, Z. A.; Ansari, S. G.; Malhotra, Bansi D.

    2015-11-01

    Nanomaterial-based photoluminescence (PL) diagnostic devices offer fast and highly sensitive detection of pesticides, DNA, and toxic agents. Here we report a label-free PL genosensor for sensitive detection of Vibrio cholerae that is based on a DNA hybridization strategy utilizing nanostructured magnesium oxide (nMgO; size >30 nm) particles. The morphology and size of the synthesized nMgO were determined by transmission electron microscopic (TEM) studies. The probe DNA (pDNA) was conjugated with nMgO and characterized by X-ray photoelectron and Fourier transform infrared spectroscopic techniques. The target complementary genomic DNA (cDNA) isolated from clinical samples of V. cholerae was subjected to DNA hybridization studies using the pDNA-nMgO complex and detection of the cDNA was accomplished by measuring changes in PL intensity. The PL peak intensity measured at 700 nm (red emission) increases with the increase in cDNA concentration. A linear range of response in the developed PL genosensor was observed from 100 to 500 ng/μL with a sensitivity of 1.306 emi/ng, detection limit of 3.133 ng/μL and a regression coefficient (R2) of 0.987. These results show that this ultrasensitive PL genosensor has the potential for applications in the clinical diagnosis of cholera.

  16. A gold nanoparticle-based label free colorimetric aptasensor for adenosine deaminase detection and inhibition assay.

    Science.gov (United States)

    Cheng, Fen; He, Yue; Xing, Xiao-Jing; Tan, Dai-Di; Lin, Yi; Pang, Dai-Wen; Tang, Hong-Wu

    2015-03-07

    A novel strategy for the fabrication of a colorimetric aptasensor using label free gold nanoparticles (AuNPs) is proposed in this work, and the strategy has been employed for the assay of adenosine deaminase (ADA) activity. The aptasensor consists of adenosine (AD) aptamer, AD and AuNPs. The design of the biosensor takes advantage of the special optical properties of AuNPs and the interaction between AuNPs and single-strand DNA. In the absence of ADA, the AuNPs are aggregated and are blue in color under appropriate salt concentration because of the grid structure of an AD aptamer when binding to AD, while in the presence of the analyte, AuNPs remain dispersed with red color under the same concentration of salt owing to ADA converting AD into inosine which has no affinity with the AD aptamer, thus allowing quantitative investigation of ADA activity. The present strategy is simple, cost-effective, selective and sensitive for ADA with a detection limit of 1.526 U L(-1), which is about one order of magnitude lower than that previously reported. In addition, a very low concentration of the inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) could generate a distinguishable response. Therefore, the AuNP-based colorimetric biosensor has great potential in the diagnosis of ADA-relevant diseases and drug screening.

  17. Machine Learning Enables Live Label-Free Phenotypic Screening in Three Dimensions.

    Science.gov (United States)

    O'Duibhir, Eoghan; Paris, Jasmin; Lawson, Hannah; Sepulveda, Catarina; Shenton, Dahlia Doughty; Carragher, Neil O; Kranc, Kamil R

    2018-01-01

    There is a large amount of information in brightfield images that was previously inaccessible by using traditional microscopy techniques. This information can now be exploited by using machine-learning approaches for both image segmentation and the classification of objects. We have combined these approaches with a label-free assay for growth and differentiation of leukemic colonies, to generate a novel platform for phenotypic drug discovery. Initially, a supervised machine-learning algorithm was used to identify in-focus colonies growing in a three-dimensional (3D) methylcellulose gel. Once identified, unsupervised clustering and principle component analysis of texture-based phenotypic profiles were applied to group similar phenotypes. In a proof-of-concept study, we successfully identified a novel phenotype induced by a compound that is currently in clinical trials for the treatment of leukemia. We believe that our platform will be of great benefit for the utilization of patient-derived 3D cell culture systems for both drug discovery and diagnostic applications.

  18. Label-Free Imaging of Umbilical Cord Tissue Morphology and Explant-Derived Cells

    Science.gov (United States)

    Paesen, Rik; Gyselaers, Wilfried; Stinissen, Piet

    2016-01-01

    In situ detection of MSCs remains difficult and warrants additional methods to aid with their characterization in vivo. Two-photon confocal laser scanning microscopy (TPM) and second harmonic generation (SHG) could fill this gap. Both techniques enable the detection of cells and extracellular structures, based on intrinsic properties of the specific tissue and intracellular molecules under optical irradiation. TPM imaging and SHG imaging have been used for label-free monitoring of stem cells differentiation, assessment of their behavior in biocompatible scaffolds, and even cell tracking in vivo. In this study, we show that TPM and SHG can accurately depict the umbilical cord architecture and visualize individual cells both in situ and during culture initiation, without the use of exogenously applied labels. In combination with nuclear DNA staining, we observed a variance in fluorescent intensity in the vessel walls. In addition, antibody staining showed differences in Oct4, αSMA, vimentin, and ALDH1A1 expression in situ, indicating functional differences among the umbilical cord cell populations. In future research, marker-free imaging can be of great added value to the current antigen-based staining methods for describing tissue structures and for the identification of progenitor cells in their tissue of origin. PMID:27746820

  19. Absolute sizing and label-free identification of extracellular vesicles by flow cytometry.

    Science.gov (United States)

    van der Pol, Edwin; de Rond, Leonie; Coumans, Frank A W; Gool, Elmar L; Böing, Anita N; Sturk, Auguste; Nieuwland, Rienk; van Leeuwen, Ton G

    2018-01-05

    Blood contains extracellular vesicles (EVs), which are biological nanoparticles with clinical applications. In blood plasma, EVs are outnumbered by similar-sized lipoprotein particles (LPs), leading to controversial data such as non-specific binding of antibodies to LPs. Flow cytometry is a clinically applicable technique to characterize single EVs in body fluids. However, flow cytometry data have arbitrary units, impeding standardization, data comparison, and data interpretation, such as differentiation between EVs and LPs. Here we present a new method, named flow cytometry scatter ratio (Flow-SR), to relate the ambiguous light scattering signals of flow cytometry to the diameter and refractive index (RI) of single nanoparticles between 200-500 nm in diameter. Flow-SR enables label-free differentiation between EVs and LPs and improves data interpretation and comparison. Because Flow-SR is easy to implement, widely applicable, and more accurate and faster than existing techniques to size nanoparticles in suspension, Flow-SR has numerous applications in nanomedicine. Copyright © 2018 The Author. Published by Elsevier Inc. All rights reserved.

  20. Theoretical analysis of bimetallic nanorod dimer biosensors for label-free molecule detection

    Science.gov (United States)

    Das, Avijit; Talukder, Muhammad Anisuzzaman

    2018-02-01

    In this work, we theoretically analyze a gold (Au) core within silver (Ag) shell (Au@Ag) nanorod dimer biosensor for label-free molecule detection. The incident light on an Au@Ag nanorod strongly couples to localized surface plasmon modes, especially around the tip region. The field enhancement around the tip of a nanorod or between the tips of two longitudinally aligned nanorods as in a dimer can be exploited for sensitive detection of biomolecules. We derive analytical expressions for the interactions of an Au@Ag nanorod dimer with the incident light. We also study the detail dynamics of an Au@Ag nanorod dimer with the incident light computationally using finite difference time domain (FDTD) technique when core-shell ratio, relative position of the nanorods, and angle of incidence of light change. We find that the results obtained using the developed analytical model match well with that obtained using FDTD simulations. Additionally, we investigate the sensitivity of the Au@Ag nanorod dimer, i.e., shift in the resonance wavelength, when a target biomolecule such as lysozyme (Lys), human serum albumin (HSA), anti-biotin (Abn), human catalase (CAT), and human fibrinogen (Fb) protein molecules are attached to the tips of the nanorods.

  1. A Label-Free Aptamer-Based Fluorescent Assay for Cadmium Detection

    Directory of Open Access Journals (Sweden)

    Yunxia Luan

    2016-12-01

    Full Text Available Selective detection of ultratrace amounts of cadmium (Cd2+ is extremely important for food safety and environmental monitoring because of its toxicity and widespread use. In this work, we developed a facile, rapid, sensitive, and highly selective method for the detection of Cd2+ based on a label-free aptasensor using an unmodified double-stranded deoxyribonucleic acid-specific dye (PicoGreen. The linear range was 0.10–100 µg/mL, and the detection limit (0.038 ng/mL was lower than the guideline from the World Health Organization for Cd2+ in drinking water (3 ng/mL. The sensor exhibited excellent selectivity towards Cd2+ ions. We tested the aptasensor in application to a series of real water samples spiked with different concentrations of Cd2+. Compared with atomic absorption spectrometry, the results showed good tolerance to the matrix effect. The developed approach shows great potential for on-site and high-throughput analysis in routine monitoring.

  2. Low cost label-free live cell imaging for biological samples

    Science.gov (United States)

    Seniya, C.; Towers, C. E.; Towers, D. P.

    2017-02-01

    This paper reports the progress to develop a practical phase measuring microscope offering new capabilities in terms of phase measurement accuracy and quantification of cell:cell interactions over the longer term. A novel, low cost phase interference microscope for imaging live cells (label-free) is described. The method combines the Zernike phase contrast approach with a dual mirror design to enable phase modulation between the scattered and un-scattered optical fields. Two designs are proposed and demonstrated, one of which retains the common path nature of Zernike's original microscopy concept. In both setups the phase shift is simple to control via a piezoelectric driven mirror in the back focal plane of the imaging system. The approach is significantly cheaper to implement than those based on spatial light modulators (SLM) at approximately 20% of the cost. A quantitative assessment of the performance of a set of phase shifting algorithms is also presented, specifically with regard to broad bandwidth illumination in phase contrast microscopy. The simulation results show that the phase measurement accuracy is strongly dependent on the algorithm selected and the optical path difference in the sample.

  3. A Label-Free Immunosensor for Ultrasensitive Detection of Ketamine Based on Quartz Crystal Microbalance

    Directory of Open Access Journals (Sweden)

    Ya Yang

    2015-04-01

    Full Text Available In this study, we have developed a label-free immunosensor with the variation of resonance frequency (Δf of a quartz crystal microbalance (QCM as readout signal for ultrasensitive detection of Ketamine (KT. An optimized strategy for immobilization of KT antibody on the surface of the QCM chip was implemented via the self-assembly modification of 3-mercaptopropionic acid, and then activated with 1-ethyl-3- (3-dimethylaminoprophl carbodiimide and n-hydroxysuccinimide. The specific affinity between the antibody and the antigen ensured a selective response toward KT. The Δf linearly related to the concentration of KT in the range of 1 to 40 pg/mL, with a detection limit of 0.86 pg/mL (S/N = 3. The obtained immunosensor was applied to detect the KT in spiked human urine without any pretreatment but dilution with recoveries from 91.8% to 108%. The developed sensor is promising to perform the portable or on-spot KT detection in clinic or forensic cases.

  4. Review of Transducer Principles for Label-Free Biomolecular Interaction Analysis

    Directory of Open Access Journals (Sweden)

    Janos Vörös

    2011-07-01

    Full Text Available Label-free biomolecular interaction analysis is an important technique to study the chemical binding between e.g., protein and protein or protein and small molecule in real-time. The parameters obtained with this technique, such as the affinity, are important for drug development. While the surface plasmon resonance (SPR instruments are most widely used, new types of sensors are emerging. These developments are generally driven by the need for higher throughput, lower sample consumption or by the need of complimentary information to the SPR data. This review aims to give an overview about a wide range of sensor transducers, the working principles and the peculiarities of each technology, e.g., concerning the set-up, sensitivity, sensor size or required sample volume. Starting from optical technologies like the SPR and waveguide based sensors, acoustic sensors like the quartz crystal microbalance (QCM and the film bulk acoustic resonator (FBAR, calorimetric and electrochemical sensors are covered. Technologies long established in the market are presented together with those newly commercially available and with technologies in the early development stage. Finally, the commercially available instruments are summarized together with their sensitivity and the number of sensors usable in parallel and an outlook for potential future developments is given.

  5. Label-free detection of sex determining region Y (SRY) via capacitive biosensor

    KAUST Repository

    Sivashankar, Shilpa

    2016-10-20

    In this work, we present for the first time, the use of a simple fractal capacitive biosensor for the quantification and detection of sex-determining region Y (SRY) genes. This section of genetic code, which is found on the Y chromosome, finds importance for study as it causes fetuses to develop characteristics of male sex-like gonads when a mutation occurs. It is also an important genetic code in men, and disorders involving the SRY gene can cause infertility and sexual malfunction that lead to a variety of gene mutational disorders. We have therefore designed silicon-based, label-free fractal capacitive biosensors to quantify various proteins and genes. We take advantage of a good dielectric material, Parylene C for enhancing the performance of the sensors. We have integrated these sensors with a simple microchannel for easy handling of fluids on the detection area. The read-out value of an Agilent LCR meter used to measure capacitance of the sensor at a frequency of 1 MHz determined gene specificity and gene quantification. These data revealed that the capacitance measurement of the capacitive biosensor for the SRY gene depended on both the target and the concentration of DNA. The experimental outcomes in the present study can be used to detect DNA and its variations in crucial fields that have a great impact on our daily lives, such as clinical and veterinary diagnostics, industrial and environmental testing and forensic sciences.

  6. Thioflavin T behaves as an efficient fluorescent ligand for label-free ATP aptasensor.

    Science.gov (United States)

    Wang, Huihui; Peng, Pai; Liu, Shuangna; Li, Tao

    2016-11-01

    Here, we for the first time demonstrated thioflavin T (ThT) as an efficient fluorescent ligand for 27-mer ATP-binding aptamer (ABA27), providing a novel signal readout mode for label-free selective ATP detection. ABA27 can promote the fluorescence emission of ThT with an unprecedentedly high efficiency, attributed to the specific structure of ABA27 rather than the G-tracts. Polyacrylamide gel electrophoresis, fluorescence spectroscopy, and fluorometric titration reveal that ThT interacts with ABA27 with a lower binding affinity (Kd ~89 μM) than ATP, which allows ATP to easily compete with ThT for the DNA binder. In the presence of ThT, adding ATP induces ABA27 to undergo a structural change, thereby not favoring the binding to ThT, verified by circular dichroism and UV-Vis absorption spectroscopy. As a result, the fluorescence intensity of ThT decreases dramatically, enabling the sensitive detection of ATP with high selectivity over other analogs. Such a sensing strategy may make ThT able to serve as a facile signal reporter for DNA nanomechanical devices fueled with ATP. Graphical Abstract The principle of the displacement of ThT by ATP.

  7. Applying a Targeted Label-free Approach using LC-MS AMT Tags to Evaluate Changes in Protein Phosphorylation Following Phosphatase Inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Feng; Jaitly, Navdeep; Jayachandran, Hemalatha; Lou, Quanzhou; Monroe, Matthew E.; Du, Xiuxia; Gritsenko, Marina A.; Zhang, Rui; Anderson, David J.; Purvine, Samuel O.; Adkins, Joshua N.; Moore, Ronald J.; Mottaz, Heather M.; Ding, Shi-Jian; Lipton, Mary S.; Camp, David G.; Udseth, Harold R.; Smith, Richard D.; Rossie, Sandra S.

    2007-10-12

    To identify phosphoproteins regulated by the phosphoprotein phosphatase (PPP) family of S/T phosphatases, we performed a large-scale characterization of changes in protein phosphorylation on extracts from HeLa cells treated with or without calyculin A, a potent PPP enzyme inhibitor. A label-free comparative Phosphoproteomics approach using immobilized metal ion affinity chromatography and targeted tandem mass spectrometry was employed to discover and identify signatures based upon distinctive changes in abundance. Overall, 232 proteins were identified as either direct or indirect targets for PPP enzyme regulation. Most of the present identifications represent novel PPP enzyme targets at the level of both phosphorylation site and protein. These include phosphorylation sites within signaling proteins such as p120 Catenin, A Kinase Anchoring Protein 8, JunB, and Type II Phosphatidyl Inositol 4 Kinase. These data can be used to define underlying signaling pathways and events regulated by the PPP family of S/T phosphatases.

  8. A label-free fluorescence biosensor for highly sensitive detection of lectin based on carboxymethyl chitosan-quantum dots and gold nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ziping; Liu, Hua; Wang, Lei; Su, Xingguang, E-mail: suxg@jlu.edu.cn

    2016-08-17

    In this work, we report a novel label-free fluorescence “turn off-on” biosensor for lectin detection. The highly sensitive and selective sensing system is based on the integration of carboxymethyl chitosan (CM-CHIT), CuInS{sub 2} quantum dots (QDs) and Au nanoparticles (NPs). Firstly, CuInS{sub 2} QDs featuring carboxyl groups were directly synthesized via a hydrothermal synthesis method. Then, the carboxyl groups on the CuInS{sub 2} QDs surface were interacted with the amino groups (−NH{sub 2}), carboxyl groups (−COOH) and hydroxyl groups (−OH) within CM-CHIT polymeric chains via electrostatic interactions and hydrogen bonding to form CM-CHIT-QDs assemblies. Introduction of Au NPs could quench the fluorescence of CM-CHIT-QDs through electron and energy transfer. In the presence of lectin, lectin could bind exclusively with CM-CHIT-QDs by means of specific multivalent carbohydrate-protein interaction. Thus, the electron and energy transfer process between CM-CHIT-QDs and Au NPs was inhibited, and as a result, the fluorescence of CM-CHIT-QDs was effectively “turned on”. Under the optimum conditions, there was a good linear relationship between the fluorescence intensity ratio I/I{sub 0} (I and I{sub 0} were the fluorescence intensity of CM-CHIT-QDs-Au NPs in the presence and absence of lectin, respectively) and lectin concentration in the range of 0.2–192.5 nmol L{sup −1}, And the detection limit could be down to 0.08 nmol L{sup −1}. Furthermore, the proposed biosensor was employed for the determination of lectin in fetal bovine serum samples with satisfactory results. - Graphical abstract: A label-free fluorescence biosensor for highly sensitive detection of lectin based on the integration of carboxymethyl chitosan, CuInS{sub 2} quantum dots and gold nanoparticles. - Highlights: • A label-free near-infrared fluorescence “turn off-on” biosensor for detection of lectin was established. • The highly sensitive biosensor was based on the

  9. Direct numerical simulation of noninvasive channel healing in electrical field

    KAUST Repository

    Wang, Yi

    2017-11-25

    Noninvasive channel healing is a new idea to repair the broken pipe wall, using external electric fields to drive iron particles to the destination. The repair can be done in the normal operation of the pipe flow without any shutdown of the pipeline so that this method can be a potentially efficient and safe technology of pipe healing. However, the real application needs full knowledge of healing details. Numerical simulation is an effective method. Thus, in this research, we first established a numerical model for noninvasive channel healing technology to represent fluid–particle interaction. The iron particles can be attached to a cracking area by external electrostatic forces or can also be detached by mechanical forces from the fluid. When enough particles are permanently attached on the cracking area, the pipe wall can be healed. The numerical criterion of the permanent attachment is discussed. A fully three-dimensional finite difference framework of direct numerical simulation is established and applied to different cases to simulate the full process of channel healing. The impact of Reynolds number and particle concentration on the healing process is discussed. This numerical investigation provides valuable reference and tools for further simulation of real pipe healing in engineering.

  10. Direction and Policies Needed to Support Hybrid Electric Car Research

    OpenAIRE

    Ridwan Arief Subekti; Agus Hartanto; Vita Susanti

    2012-01-01

    The rising number of vehicles over the years has driven the increase of air pollution and fuel consumption. One of the solutions to overcome this problem is using hybrid electric car because it is environmentally friendly and efficient in fuel consumption. LIPI has conducted electric car research since 1997, but there were so many problems in its development that electric car can not be developed into a national industry scale. Therefore, it is important to conduct a study that maps the probl...

  11. The development of a silica nanoparticle-based label-free DNA biosensor

    Science.gov (United States)

    Kell, Arnold J.; Pagé, Lilianne; Tan, Sophie; Charlebois, Isabelle; Boissinot, Maurice; Leclerc, Mario; Simard, Benoit

    2011-09-01

    A silica nanoparticle-based DNA biosensor capable of detecting Bacillus anthracis bacteria through the use of unlabelled ss-oligonucleotides has been developed. The biosensor makes use of the optical changes that accompany a nanoparticle-immobilized cationic conjugated polymer (polythiophene) interacting with single-stranded vs. hybridized oligonucleotides, where a fluorescence signal appears only when hybridized DNA is present (i.e. only when the ss-oligonucleotide interacting with the polymer has hybridized with its complement). In order to enhance the sensitivity of the biosensor, two different nanoparticle architectures were developed and used to elucidate how the presence of neighboring fluorophores on the nanoparticle surface affects Förster-resonant energy transfer (FRET) between the polythiophene/oligonucleotide complex (FRET donor) and the fluorophores (FRET acceptors). We demonstrate that the silica nanoparticle-based FRET platform lowers the limit of detection at least 10-fold in comparison to the polythiophene itself, and allows the detection of ~2 × 10-12 moles of ss-oligonucleotide in a 100 μL sample with a standard fluorimeter (i.e. has a limit of detection of ~2 nM ssDNA). Such nanoparticle-based biosensor platforms are beneficial because of the robustness and stability inherent to their covalent assembly and they provide a valuable new tool that may allow for the sensitive, label-free detection (the target DNA that produces the fluorescence signal is unlabelled) without the use of polymerase chain reaction.A silica nanoparticle-based DNA biosensor capable of detecting Bacillus anthracis bacteria through the use of unlabelled ss-oligonucleotides has been developed. The biosensor makes use of the optical changes that accompany a nanoparticle-immobilized cationic conjugated polymer (polythiophene) interacting with single-stranded vs. hybridized oligonucleotides, where a fluorescence signal appears only when hybridized DNA is present (i.e. only when

  12. Rapid label-free identification of mixed bacterial infections by surface plasmon resonance

    Directory of Open Access Journals (Sweden)

    Fu Weiling

    2011-06-01

    Full Text Available Abstract Background Early detection of mixed aerobic-anaerobic infection has been a challenge in clinical practice due to the phenotypic changes in complex environments. Surface plasmon resonance (SPR biosensor is widely used to detect DNA-DNA interaction and offers a sensitive and label-free approach in DNA research. Methods In this study, we developed a single-stranded DNA (ssDNA amplification technique and modified the traditional SPR detection system for rapid and simultaneous detection of mixed infections of four pathogenic microorganisms (Pseudomonas aeruginosa, Staphylococcus aureus, Clostridium tetani and Clostridium perfringens. Results We constructed the circulation detection well to increase the sensitivity and the tandem probe arrays to reduce the non-specific hybridization. The use of 16S rDNA universal primers ensured the amplification of four target nucleic acid sequences simultaneously, and further electrophoresis and sequencing confirmed the high efficiency of this amplification method. No significant signals were detected during the single-base mismatch or non-specific probe hybridization (P 2 values of >0.99. The lowest detection limits were 0.03 nM for P. aeruginosa, 0.02 nM for S. aureus, 0.01 nM for C. tetani and 0.02 nM for C. perfringens. The SPR biosensor had the same detection rate as the traditional culture method (P Conclusions Our method can rapidly and accurately identify the mixed aerobic-anaerobic infection, providing a reliable alternative to bacterial culture for rapid bacteria detection.

  13. Time resolved and label free monitoring of extracellular metabolites by surface enhanced Raman spectroscopy.

    Directory of Open Access Journals (Sweden)

    Victoria Shalabaeva

    Full Text Available Metabolomics is an emerging field of cell biology that aims at the comprehensive identification of metabolite levels in biological fluids or cells in a specific functional state. Currently, the major tools for determining metabolite concentrations are mass spectrometry coupled with chromatographic techniques and nuclear magnetic resonance, which are expensive, time consuming and destructive for the samples. Here, we report a time resolved approach to monitor metabolite dynamics in cell cultures, based on Surface Enhanced Raman Scattering (SERS. This method is label-free, easy to use and provides the opportunity to simultaneously study a broad range of molecules, without the need to process the biological samples. As proof of concept, NIH/3T3 cells were cultured in vitro, and the extracellular medium was collected at different time points to be analyzed with our engineered SERS substrates. By identifying individual peaks of the Raman spectra, we showed the simultaneous detection of several components of the conditioned medium, such as L-tyrosine, L-tryptophan, glycine, L-phenylalanine, L-histidine and fetal bovine serum proteins, as well as their intensity changes during time. Furthermore, analyzing the whole Raman data set with the Principal Component Analysis (PCA, we demonstrated that the Raman spectra collected at different days of culture and clustered by similarity, described a well-defined trajectory in the principal component plot. This approach was then utilized to determine indirectly the functional state of the macrophage cell line Raw 264.7, stimulated with the lipopolysaccharide (LPS for 24 hours. The collected spectra at different time points, clustered by the PCA analysis, followed a well-defined trajectory, corresponding to the functional change of cells toward the activated pro-inflammatory state induced by the LPS. This study suggests that our engineered SERS surfaces can be used as a versatile tool both for the characterization

  14. A label-free photoelectrochemical cocaine aptasensor based on an electropolymerized ruthenium-intercalator complex

    International Nuclear Information System (INIS)

    Haddache, Fatima; Le Goff, Alan; Spinelli, Nicolas; Gairola, Priyanka; Gorgy, Karine; Gondran, Chantal; Defrancq, Eric; Cosnier, Serge

    2016-01-01

    Highlights: • Electrodes were modified by an electrogenerated Ru(II) complex which demonstrates photosensitive properties and intercalating properties towards the stem-loop base pairing domain of cocaine aptamers. • Cocaine aptamers were immobilized as mono-and double-fragment which showed different behaviour towards photocurrent generation. • The binding of aptamer could be followed by photelectrochemistry and modelized using a Langmuir-Freundlich isotherm. • Using the double-fragment aptamer, a label-free photoelectrochemical aptasensor was designed, exhibiting a LOD of 10 nmol L −1 and linear range of 1 10 −8 –5 10 −4 mol L −1 . - Abstract: A photoelectrode was designed by electrodeposition of a pyrrole monomer modified with a polypyridyl Ru(II) complex bearing benzo[i]dipyrido-[3,2-a:2′.3′-c]phenazine (dppn) ligand. Owing to the intercalating properties of these immobilized complexes towards DNA double helix, cocaine aptamer was immobilized on the modified electrodes thanks to its stem-loop configuration in order to design a photoelectrochemical cocaine aptasensor. Especially using a double-fragment aptamer strategy, the binding of cocaine and the formation of the aptamer/cocaine complex was successfully observed and modeled by a Langmuir-Freundlich isotherm, giving access to an apparent dissociation constant K d of 3.8 mmol L −1 . The photoelectrochemical aptasensor exhibits a LOD of 10 nmol L −1 and linear range of 1 10 −8 –5 10 −4 mol L −1 .

  15. MoS₂ field-effect transistor for next-generation label-free biosensors.

    Science.gov (United States)

    Sarkar, Deblina; Liu, Wei; Xie, Xuejun; Anselmo, Aaron C; Mitragotri, Samir; Banerjee, Kaustav

    2014-04-22

    Biosensors based on field-effect transistors (FETs) have attracted much attention, as they offer rapid, inexpensive, and label-free detection. While the low sensitivity of FET biosensors based on bulk 3D structures has been overcome by using 1D structures (nanotubes/nanowires), the latter face severe fabrication challenges, impairing their practical applications. In this paper, we introduce and demonstrate FET biosensors based on molybdenum disulfide (MoS2), which provides extremely high sensitivity and at the same time offers easy patternability and device fabrication, due to its 2D atomically layered structure. A MoS2-based pH sensor achieving sensitivity as high as 713 for a pH change by 1 unit along with efficient operation over a wide pH range (3-9) is demonstrated. Ultrasensitive and specific protein sensing is also achieved with a sensitivity of 196 even at 100 femtomolar concentration. While graphene is also a 2D material, we show here that it cannot compete with a MoS2-based FET biosensor, which surpasses the sensitivity of that based on graphene by more than 74-fold. Moreover, we establish through theoretical analysis that MoS2 is greatly advantageous for biosensor device scaling without compromising its sensitivity, which is beneficial for single molecular detection. Furthermore, MoS2, with its highly flexible and transparent nature, can offer new opportunities in advanced diagnostics and medical prostheses. This unique fusion of desirable properties makes MoS2 a highly potential candidate for next-generation low-cost biosensors.

  16. A label free aptasensor for Ochratoxin A detection in cocoa beans: An application to chocolate industries

    International Nuclear Information System (INIS)

    Mishra, Rupesh K.; Hayat, Akhtar; Catanante, Gaëlle; Ocaña, Cristina; Marty, Jean-Louis

    2015-01-01

    Contamination of food by mycotoxin occurs in minute/trace quantities. Nearly 92.5% of the cocoa samples present Ochratoxin A (OTA) levels at trace quantity. Hence, there is a necessity for a highly sensitive and selective device that can detect and quantify these organic toxins in various matrices such as cocoa beans. This work reports for the first time, a facile and label-free electrochemical impedimetric aptasensor for rapid detection and quantitation of OTA in cocoa beans. The developed aptasensor was constructed based on the diazonium-coupling reaction mechanism for the immobilization of anti-OTA-aptamer on screen printed carbon electrodes (SPCEs). The aptasensor exhibited a very good limit of detection (LOD) as low as 0.15 ng/mL, with added advantages of good selectivity and reproducibility. The increase in electron transfer resistance was linearly proportional to the OTA concentration in the range 0.15–2.5 ng/mL, with an acceptable recovery percentage (91–95%, RSD = 4.8%) obtained in cocoa samples. This work can facilitate a general model for the detection of OTA in cocoa beans based on the impedimetric aptasensor. The analysis can be performed onsite with pre-constructed and aptamer modified electrodes employing a portable EIS set up. - Highlights: • Simple and facile method to detect OTA. • The aptasensor exhibited a very good limit of detection (LOD) as low as 0.15 ng/mL. • The first report on OTA detection in cocoa beans using impedimetric aptasensor

  17. Label-free histamine detection with nanofluidic diodes through metal ion displacement mechanism.

    Science.gov (United States)

    Ali, Mubarak; Ramirez, Patricio; Duznovic, Ivana; Nasir, Saima; Mafe, Salvador; Ensinger, Wolfgang

    2017-02-01

    We design and characterize a nanofluidic device for the label-free specific detection of histamine neurotransmitter based on a metal ion displacement mechanism. The sensor consists of an asymmetric polymer nanopore fabricated via ion track-etching technique. The nanopore sensor surface having metal-nitrilotriacetic (NTA-Ni 2+ ) chelates is obtained by covalent coupling of native carboxylic acid groups with N α ,N α -bis(carboxymethyl)-l-lysine (BCML), followed by exposure to Ni 2+ ion solution. The BCML immobilization and subsequent Ni 2+ ion complexation with NTA moieties change the surface charge concentration, which has a significant impact on the current-voltage (I-V) curve after chemical modification of the nanopore. The sensing mechanism is based on the displacement of the metal ion from the NTA-Ni 2+ chelates. When the modified pore is exposed to histamine solution, the Ni 2+ ion in NTA-Ni 2+ chelate recognizes histamine through a metal ion coordination displacement process and formation of stable Ni-histamine complexes, leading to the regeneration of metal-free NTA groups on the pore surface, as shown in the current-voltage characteristics. Nanomolar concentrations of the histamine in the working electrolyte can be detected. On the contrary, other neurotransmitters such as glycine, serotonin, gamma-aminobutyric acid, and dopamine do not provoke significant changes in the nanopore electronic signal due to their inability to displace the metal ion and form a stable complex with Ni 2+ ion. The nanofluidic sensor exhibits high sensitivity, specificity and reusability towards histamine detection and can then be used to monitor the concentration of biological important neurotransmitters. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. A label free aptasensor for Ochratoxin A detection in cocoa beans: An application to chocolate industries

    Energy Technology Data Exchange (ETDEWEB)

    Mishra, Rupesh K. [IMAGES, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860 (France); Hayat, Akhtar [IMAGES, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860 (France); Interdisciplinary Research Centre in Biomedical Materials (IRCBM), COMSATS Institute of Information Technology (CIIT), Lahore 54000 (Pakistan); Catanante, Gaëlle; Ocaña, Cristina [IMAGES, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860 (France); Marty, Jean-Louis, E-mail: jlmarty@univ-perp.fr [IMAGES, Université De Perpignan Via Domitia, 52 Avenue Paul Alduy, Perpignan Cedex 66860 (France)

    2015-08-19

    Contamination of food by mycotoxin occurs in minute/trace quantities. Nearly 92.5% of the cocoa samples present Ochratoxin A (OTA) levels at trace quantity. Hence, there is a necessity for a highly sensitive and selective device that can detect and quantify these organic toxins in various matrices such as cocoa beans. This work reports for the first time, a facile and label-free electrochemical impedimetric aptasensor for rapid detection and quantitation of OTA in cocoa beans. The developed aptasensor was constructed based on the diazonium-coupling reaction mechanism for the immobilization of anti-OTA-aptamer on screen printed carbon electrodes (SPCEs). The aptasensor exhibited a very good limit of detection (LOD) as low as 0.15 ng/mL, with added advantages of good selectivity and reproducibility. The increase in electron transfer resistance was linearly proportional to the OTA concentration in the range 0.15–2.5 ng/mL, with an acceptable recovery percentage (91–95%, RSD = 4.8%) obtained in cocoa samples. This work can facilitate a general model for the detection of OTA in cocoa beans based on the impedimetric aptasensor. The analysis can be performed onsite with pre-constructed and aptamer modified electrodes employing a portable EIS set up. - Highlights: • Simple and facile method to detect OTA. • The aptasensor exhibited a very good limit of detection (LOD) as low as 0.15 ng/mL. • The first report on OTA detection in cocoa beans using impedimetric aptasensor.

  19. A Label-Free Microelectrode Array Based on One-Step Synthesis of Chitosan–Multi-Walled Carbon Nanotube–Thionine for Ultrasensitive Detection of Carcinoembryonic Antigen

    Directory of Open Access Journals (Sweden)

    Huiren Xu

    2016-07-01

    Full Text Available Carcinoembryonic antigen (CEA has been an extensively used tumor marker responsible for clinical early diagnosis of cervical carcinomas, and pancreatic, colorectal, gastric and lung cancer. Combined with micro-electro mechanical system (MEMS technology, it is important to develop a novel immune microelectrode array (MEA not only for rapid analysis of serum samples, but also for cell detection in vitro and in vivo. In this work, we depict a simple approach to modify chitosan–multi-walled carbon nanotubes–thionine (CS–MWCNTs–THI hybrid film through one-step electrochemical deposition and the CS-MWCNTs-THI hybrid films are successfully employed to immobilize anti-CEA for fabricating simple, label-free, and highly sensitive electro-chemical immune MEAs. The detection principle of immune MEA was based on the fact that the increasing formation of the antigen-antibody immunocomplex resulted in the decreased response currents and the relationship between the current reductions with the corresponding CEA concentrations was directly proportional. Experimental results indicated that the label-free MEA had good selectivity and the limit of detection for CEA is 0.5 pg/mL signal to noise ratio (SNR = 3. A linear calibration plot for the detection of CEA was obtained in a wide concentration range from 1 pg/mL to 100 ng/mL (r = 0.996. This novel MEA has potential applications for detecting CEA for the research on cancer cells and cancer tissue slices as well as for effective early diagnosis.

  20. Direction and Policies Needed to Support Hybrid Electric Car Research

    Directory of Open Access Journals (Sweden)

    Ridwan Arief Subekti

    2012-07-01

    Full Text Available The rising number of vehicles over the years has driven the increase of air pollution and fuel consumption. One of the solutions to overcome this problem is using hybrid electric car because it is environmentally friendly and efficient in fuel consumption. LIPI has conducted electric car research since 1997, but there were so many problems in its development that electric car can not be developed into a national industry scale. Therefore, it is important to conduct a study that maps the problems and finds the solutions to prevent the same failure of electric car commercialization process from happening to hybrid electric car . This study was done by collecting and analyzing the primary and secondary data through interviews, discussing electric hybrid car with stakeholders, and examining earlier study results and regulations. Based on this study, several policies to support sustainability research of hybrid electric car were proposed. Some recommendations were the making of national roadmap and regulation for the usage of hybrid electric car on the road. For policy makers at LIPI, a research focus, research coordination, and pre-commercialization program were recommended.

  1. Direct Carbon Fuel Cells: Converting Waste to Electricity

    National Research Council Canada - National Science Library

    Wolk, Ronald H; Lux, Scott; Gelber, Stacy; Holcomb, Franklin H

    2007-01-01

    ... U.S. Army Solid Waste Annual Reporting System (SWARS) database, then estimated the amount of electricity that DCFC technologies could generate from those wastes, and finally compared those amounts with the current average annual electrical loads at the 10 largest Army installations.

  2. The direct conversion of heat into electricity in reactors

    International Nuclear Information System (INIS)

    Devin, B.; Bliaux, J.; Lesueur, R.

    1964-01-01

    The direct conversion of heat into electricity by thermionic emission in an atomic reactor has been studied with the triple aim of its utilisation: as an energy source for a space device, at the head of a conventional conversion system in power installations, or finally in association with the thermoelectric conversion in very low power installations. The laboratory experiments were mainly orientated towards the electron extraction of metals and compounds and their behaviour at high temperatures. Converters furnishing up to 50 amps at 0. 4 volts with an efficiency close to 10 p. 100 have been constructed in the laboratory; the emitters were heated by electron bombardment and were composed of tungsten covered with an uranium carbide deposit or molybdenum covered with cesium. The main aspects of the coupling between the converter and the reactor have been covered from the point of view of electronics: the influence of the mismatching of the load on the temperature of the emitter and the influence of thermal flux density on the temperature of the emitter and the stability of the converter. Converters using uranium carbide as the electron emitter have been tested in reactors. Tests have been made under dynamic conditions in order to determine the dynamic characteristics. The load matching curves have been constructed and the overall performances of several cells coupled in such a way as to form a reactor rod have been deduced. This information is fundamental to the design of a control system for a thermionic conversion reactor. The problems associated with the reliability of thermionic converters connected in series in the same reactor rod have been examined theoretically. Finally, the absorption isotherms have been drawn at the ambient temperatures for krypton and xenon on activated carbon with the aim of investigating the escape of fission products in a converter. (author) [fr

  3. Direct Electrical Current Reduces Bacterial and Yeast Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Maria Ruiz-Ruigomez

    2016-01-01

    Full Text Available New strategies are needed for prevention of biofilm formation. We have previously shown that 24 hr of 2,000 µA of direct current (DC reduces Staphylococcus epidermidis biofilm formation in vitro. Herein, we examined the effect of a lower amount of DC exposure on S. epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Propionibacterium acnes, and Candida albicans biofilm formation. 12 hr of 500 µA DC decreased S. epidermidis, S. aureus, E. coli, and P. aeruginosa biofilm formation on Teflon discs by 2, 1, 1, and 2 log10 cfu/cm2, respectively (p<0.05. Reductions in S. epidermidis, S. aureus, and E. coli biofilm formation were observed with as few as 12 hr of 200 µA DC (2, 2 and 0.4 log10 cfu/cm2, resp.; a 1 log10 cfu/cm2 reduction in P. aeruginosa biofilm formation was observed at 36 hr. 24 hr of 500 µA DC decreased C. albicans biofilm formation on Teflon discs by 2 log10 cfu/cm2. No reduction in P. acnes biofilm formation was observed. 1 and 2 log10 cfu/cm2 reductions in E. coli and S. epidermidis biofilm formation on titanium discs, respectively, were observed with 12 hr of exposure to 500 µA. Electrical current is a potential strategy to reduce biofilm formation on medical biomaterials.

  4. Elastic and inelastic light scattering spectroscopy and its possible use for label-free brain tumor typing.

    Science.gov (United States)

    Ostertag, Edwin; Stefanakis, Mona; Rebner, Karsten; Kessler, Rudolf W

    2017-11-01

    This paper presents an approach for label-free brain tumor tissue typing. For this application, our dual modality microspectroscopy system combines inelastic Raman scattering spectroscopy and Mie elastic light scattering spectroscopy. The system enables marker-free biomedical diagnostics and records both the chemical and morphologic changes of tissues on a cellular and subcellular level. The system setup is described and the suitability for measuring morphologic features is investigated. Graphical Abstract Bimodal approach for label-free brain tumor typing. Elastic and inelastic light scattering spectra are collected laterally resolved in one measurement setup. The spectra are investigated by multivariate data analysis for assigning the tissues to specific WHO grades according to their malignancy.

  5. Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope.

    Science.gov (United States)

    Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

    2015-11-03

    Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications.

  6. Dynamic nano-imaging of label-free living cells using electron beam excitation-assisted optical microscope

    Science.gov (United States)

    Fukuta, Masahiro; Kanamori, Satoshi; Furukawa, Taichi; Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

    2015-01-01

    Optical microscopes are effective tools for cellular function analysis because biological cells can be observed non-destructively and non-invasively in the living state in either water or atmosphere condition. Label-free optical imaging technique such as phase-contrast microscopy has been analysed many cellular functions, and it is essential technology for bioscience field. However, the diffraction limit of light makes it is difficult to image nano-structures in a label-free living cell, for example the endoplasmic reticulum, the Golgi body and the localization of proteins. Here we demonstrate the dynamic imaging of a label-free cell with high spatial resolution by using an electron beam excitation-assisted optical (EXA) microscope. We observed the dynamic movement of the nucleus and nano-scale granules in living cells with better than 100 nm spatial resolution and a signal-to-noise ratio (SNR) around 10. Our results contribute to the development of cellular function analysis and open up new bioscience applications. PMID:26525841

  7. 76 FR 64287 - Airworthiness Directives; General Electric Company CF34-10E Series Turbofan Engines

    Science.gov (United States)

    2011-10-18

    ... General Electric Company (GE) CF34-10E series turbofan engines. This proposed AD was [[Page 64288...) Applicability This AD applies to General Electric Company (GE) CF34-10E series turbofan engines, serial number...-0599; Directorate Identifier 2011-NE-19-AD] RIN 2120-AA64 Airworthiness Directives; General Electric...

  8. A label-free quantitative shotgun proteomics analysis of rice grain development

    Directory of Open Access Journals (Sweden)

    Koh Hee-Jong

    2011-09-01

    Full Text Available Abstract Background Although a great deal of rice proteomic research has been conducted, there are relatively few studies specifically addressing the rice grain proteome. The existing rice grain proteomic researches have focused on the identification of differentially expressed proteins or monitoring protein expression patterns during grain filling stages. Results Proteins were extracted from rice grains 10, 20, and 30 days after flowering, as well as from fully mature grains. By merging all of the identified proteins in this study, we identified 4,172 non-redundant proteins with a wide range of molecular weights (from 5.2 kDa to 611 kDa and pI values (from pH 2.9 to pH 12.6. A Genome Ontology category enrichment analysis for the 4,172 proteins revealed that 52 categories were enriched, including the carbohydrate metabolic process, transport, localization, lipid metabolic process, and secondary metabolic process. The relative abundances of the 1,784 reproducibly identified proteins were compared to detect 484 differentially expressed proteins during rice grain development. Clustering analysis and Genome Ontology category enrichment analysis revealed that proteins involved in the metabolic process were enriched through all stages of development, suggesting that proteome changes occurred even in the desiccation phase. Interestingly, enrichments of proteins involved in protein folding were detected in the desiccation phase and in fully mature grain. Conclusion This is the first report conducting comprehensive identification of rice grain proteins. With a label free shotgun proteomic approach, we identified large number of rice grain proteins and compared the expression patterns of reproducibly identified proteins during rice grain development. Clustering analysis, Genome Ontology category enrichment analysis, and the analysis of composite expression profiles revealed dynamic changes of metabolisms during rice grain development. Interestingly, we

  9. Label-Free Analysis of Cellular Lipid Droplet Formation by Non-Linear Microscopy

    Science.gov (United States)

    Schie, Iwan W.

    Cellular lipid droplets (LD) are cellular organelles that can be found in every cell type. Recent research indicates that cellular LD are involved in a large number of cellular metabolic functions, such as lipid metabolism, protection from lipotoxicity, protein storage and degradation, and many more. LD formation is frequently associated with adverse health effects, i.e. alcoholic and non-alcoholic fatty liver disease, diabetes type-2, as well as many cardiovascular disorders. Despite their wide presence, LDs are the least studied and most poorly understood cellular organelles. Typically, LDs are investigated using fluorescence-based techniques that require staining with exogenous fluorophores. Other techniques, e.g. biochemical assays, require the destruction of cells that prohibit the analysis of living cells. Therefore, in my thesis research I developed a novel compound fast-scanning nonlinear optical microscope equipped with the ability to also acquire Raman spectra at specific image locations. This system allows us to image label-free cellular LD formation in living cells and analyze the composition of single cellular LDs. Images can be acquired at near video-rate (˜16 frames/s). Furthermore, the system has the ability to acquire very large images of tissue of up to 7.5x15 cm2 total area by stitching together scans with dimensions of 1x1 mm2 in less than 1 minute. The system also enables the user to acquire Raman spectra from points of interest in the multiphoton images and provides chemically-specific data from sample volumes as small as 1 femtoliter. In my thesis I used this setup to determine the effects of VLDL lipolysis products on primary rat hepatocytes. By analyzing the Raman spectra and comparing the peak ratios for saturated and unsaturated fatty acid it was determined that the small cellular LD are highly saturated, while large cellular LDs contain mostly unsaturated lipids. Furthermore, I established a method to determine the specific contribution

  10. Direct Conversion of Radioisotope Energy to Electricity: Year 3 Report

    International Nuclear Information System (INIS)

    Marks Prelas; Alexey Spitsyn; Alejandro Suarez; Eric Stienfelds; Dickerson Moreno; Bia-Ling Hsu; Tushar Ghosh; Robert Tompson; Sudarshan Loyalka; Dabir Viswanath

    2003-01-01

    A new chemical reactor has been tested for Field Enhanced Diffusion by Optical Activation doping and purification of SiC, GaN and AlN films. Different conditions have been used on SiC, GaN and AlN samples including temperature variation, electrical field variation, variations in electrical current and optical activation. A 5mW (630-680) nm laser was used for optical activation. It was observed that optical activation has a major effect on ion drift rates. It was also observed that the magnitude of the electrical current also enhanced ion drift rates by a postulated current drag mechanism. I-V characteristic curves were measured to verify changes in the electrical properties of the samples SIMS was used to analyze the concentrations of impurities in the film samples before and after treatment. It has been demonstrated that the field-enhanced diffusion by optical activation method can dope and purify the films. As a result, the electrical properties of the wafers have been significantly improved during treatment especially in cases where a laser is used

  11. A highly sensitive, label-free gene sensor based on a single conducting polymer nanowire.

    Science.gov (United States)

    Kannan, Bhuvaneswari; Williams, David E; Laslau, Cosmin; Travas-Sejdic, Jadranka

    2012-05-15

    A prerequisite for exploiting sensing devices based on semiconductor nanowires is ultra-sensitive and selective direct electrical detection of biological and chemical species. Here, we constructed a transducer based on copolymer of poly(3,4,-ethylenedioxythiophene) (PEDOT) and carboxylic group functionalised PEDOT single nanowire in between gold electrodes, followed by covalent attachment of amino-modified probe oligonucleotide. The target ODNs specific to Homo sapiens Breast and ovarian cancer cells were detected at femtomolar concentration and incorporation of negative controls (non-complementary ODN) were clearly discriminated by the sensor. The ex situ measurements were performed by using two terminal device setup and the changes in the interface of the nanowire associated with the association or dissociation of ODNs were measured as change in resistance. In addition, in situ measurements were performed by utilizing scanning ion conductance microscopy to measure the change in resistance of probe modified nanowire upon addition of different concentration of target ODNs in presence of relevant buffer. The constructed, nano sensor showed highly sensitive concentration dependent resistance change. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. ELECTRIC VEHICULE – A NEW DIRECTION FOR SOCIO-ECONOMIC

    Directory of Open Access Journals (Sweden)

    ŞTEFAN CÎRSTEA

    2015-10-01

    Full Text Available Mobility is a major factor of economic growth and social development. The vehicle is the centrepiece of what mobility means and it is expected that, in future, the use of vehicle continues to grow, particularly in developing countries. This article wants to highlight the need to implement electric vehicles in contemporary society and, also, highlight some of the advantages and socio-economic benefits that such action entails. The aim of this paper is estimating the possible influence of such shift in different aspects of economics as a whole. The results underline the main advantages and disadvantages of adopting the electric vehicule.

  13. 78 FR 50320 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2013-08-19

    ... Electric Company (GE) model GEnx-2B67B turbofan engines with booster anti-ice (BAI) air duct, part number... cracks in the BAI air duct. This AD requires initial and repetitive visual inspections of the BAI air duct, removal from service of the BAI air duct if it fails inspection and, as a mandatory terminating...

  14. 77 FR 48110 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2012-08-13

    ... engine shop visit is defined as the induction of an engine into the shop for any reason. (h) Prohibition... Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of proposed... certain General Electric Company (GE) CF6-80C2 series turbofan engines. The existing AD requires...

  15. 76 FR 64844 - Airworthiness Directives; General Electric Company Turbofan Engines

    Science.gov (United States)

    2011-10-19

    ...) For the purposes of this AD, an engine shop visit is the induction of an engine into the shop after... Company Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of proposed... General Electric Company (GE) CF6-45 and CF6-50 series turbofan engines with certain low-pressure turbine...

  16. Direct numerical simulation of MHD flow with electrically conducting wall

    International Nuclear Information System (INIS)

    Satake, S.; Kunugi, T.; Naito, N.; Sagara, A.

    2006-01-01

    The 2D vortex problem and 3D turbulent channel flow are treated numerically to assess the effect of electrically conducting walls on turbulent MHD flow. As a first approximation, the twin vortex pair is considered as a model of a turbulent eddy near the wall. As the eddy approaches and collides with the wall, a high value electrical potential is induced inside the wall. The Lorentz force, associated with the potential distribution, reduces the velocity gradient in the near-wall region. When considering a fully developed turbulent channel flow, a high electrical conductivity wall was chosen to emphasize the effect of electromagnetic coupling between the wall and the flow. The analysis was performed using DNS. The results are compared with a non-MHD flow and MHD flow in the insulated channel. The mean velocity within the logarithmic region in the case of the electrically conducting wall is slightly higher than that in the non-conducting wall case. Thus, the drag is smaller compared to that in the non-conducting wall case due to a reduction of the Reynolds stress in the near wall region through the Lorentz force. This mechanism is explained via reduction of the production term in the Reynolds shear stress budget

  17. On Orbit Immuno-Based, Label-Free, White Blood Cell Counting System with MicroElectroMechanical Sensor (MEMS) Technology (OILWBCS-MEMS) Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Aurora Flight Sciences Corporation and partner, Draper Laboratory, propose to develop an on-orbit immuno-based label-free white blood cell counting system using MEMS...

  18. On Orbit Immuno-Based, Label-Free, White Blood Cell Counting System with MicroElectroMechanical Sensor (MEMS) Technology (OILWBCS-MEMS), Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Aurora Flight Sciences Corporation and our partner, Draper Laboratory, propose to develop an on orbit immuno-based, label-free, white blood cell counting system for...

  19. On Orbit Immuno-Based, Label-Free, White Blood Cell Counting System with MicroElectroMechanical Sensor (MEMS) Technology (OILWBCS-MEMS), Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Aurora Flight Sciences Corporation and partner, Draper Laboratory, propose to develop an on-orbit immuno-based label-free white blood cell counting system using MEMS...

  20. The efficiency of direct torque control for electric vehicle behavior improvement

    Directory of Open Access Journals (Sweden)

    Gasbaoui Brahim

    2011-01-01

    Full Text Available Nowadays the electric vehicle motorization control takes a great interest of industrials for commercialized electric vehicles. This paper is one example of the proposed control methods that ensure both safety and stability the electric vehicle by the means of Direct Torque Control (DTC. For motion of the vehicle the electric drive consists of four wheels: two front ones for steering and two rear ones for propulsion equipped with two induction motors, due to their lightweight simplicity and high performance. Acceleration and steering are ensured by the electronic differential, permitting safe and reliable steering at any curve. The direct torque control ensures efficiently controlled vehicle. Electric vehicle direct torque control is simulated in MATLAB SIMULINK environment. Electric vehicle (EV demonstrated satisfactory results in all type of roads constraints: straight, ramp, downhill and bends.

  1. Label-free in situ imaging of lignification in plant cell walls.

    Science.gov (United States)

    Schmidt, Martin; Perera, Pradeep; Schwartzberg, Adam M; Adams, Paul D; Schuck, P James

    2010-11-01

    Meeting growing energy demands safely and efficiently is a pressing global challenge. Therefore, research into biofuels production that seeks to find cost-effective and sustainable solutions has become a topical and critical task. Lignocellulosic biomass is poised to become the primary source of biomass for the conversion to liquid biofuels. However, the recalcitrance of these plant cell wall materials to cost-effective and efficient degradation presents a major impediment for their use in the production of biofuels and chemicals. In particular, lignin, a complex and irregular poly-phenylpropanoid heteropolymer, becomes problematic to the postharvest deconstruction of lignocellulosic biomass. For example in biomass conversion for biofuels, it inhibits saccharification in processes aimed at producing simple sugars for fermentation. The effective use of plant biomass for industrial purposes is in fact largely dependent on the extent to which the plant cell wall is lignified. The removal of lignin is a costly and limiting factor and lignin has therefore become a key plant breeding and genetic engineering target in order to improve cell wall conversion. Analytical tools that permit the accurate rapid characterization of lignification of plant cell walls become increasingly important for evaluating a large number of breeding populations. Extractive procedures for the isolation of native components such as lignin are inevitably destructive, bringing about significant chemical and structural modifications. Analytical chemical in situ methods are thus invaluable tools for the compositional and structural characterization of lignocellulosic materials. Raman microscopy is a technique that relies on inelastic or Raman scattering of monochromatic light, like that from a laser, where the shift in energy of the laser photons is related to molecular vibrations and presents an intrinsic label-free molecular "fingerprint" of the sample. Raman microscopy can afford non

  2. Field effect of screened charges: electrical detection of peptides and proteins by a thin-film resistor.

    Science.gov (United States)

    Lud, Simon Q; Nikolaides, Michael G; Haase, Ilka; Fischer, Markus; Bausch, Andreas R

    2006-02-13

    For many biotechnological applications the label-free detection of biomolecular interactions is becoming of outstanding importance. In this Article we report the direct electrical detection of small peptides and proteins by their intrinsic charges using a biofunctionalized thin-film resistor. The label-free selective and quantitative detection of small peptides and proteins is achieved using hydrophobized silicon-on-insulator (SOI) substrates functionalized with lipid membranes that incorporate metal-chelating lipids. The response of the nanometer-thin conducting silicon film to electrolyte screening effects is taken into account to determine quantitatively the charges of peptides. It is even possible to detect peptides with a single charge and to distinguish single charge variations of the analytes even in physiological electrolyte solutions. As the device is based on standard semiconductor technologies, parallelization and miniaturization of the SOI-based biosensor is achievable by standard CMOS technologies and thus a promising basis for high-throughput screening or biotechnological applications.

  3. ELECTRIC VEHICULE – A NEW DIRECTION FOR SOCIO-ECONOMIC

    OpenAIRE

    ŞTEFAN CÎRSTEA

    2015-01-01

    Mobility is a major factor of economic growth and social development. The vehicle is the centrepiece of what mobility means and it is expected that, in future, the use of vehicle continues to grow, particularly in developing countries. This article wants to highlight the need to implement electric vehicles in contemporary society and, also, highlight some of the advantages and socio-economic benefits that such action entails. The aim of this paper is estimating the possible influe...

  4. Future directions and cycles for electricity production from geothermal resources

    International Nuclear Information System (INIS)

    Michaelides, Efstathios E.

    2016-01-01

    Graphical abstract: 25% more power may be produced using binary-flashing geothermal cycles. - Highlights: • Power from geothermal power plants is continuously available and “dispatchable.” • The next generation of geothermal will include more binary plants. • Lower temperature geothermal resources will be utilized in the future. • Dry rock resources may produce a high fraction of electricity in several countries. - Abstract: Geothermal power production is economically competitive and capable to produce a high percentage of the electric power demand in several countries. The currently operating geothermal power plants utilize water from an aquifer at relatively higher temperatures and produce power using dry steam, flashing or binary cycles. A glance at the map of the global geothermal resources proves that there is a multitude of sites, where the aquifer temperature is lower. There are also many geothermal resources where a high geothermal gradient exists in the absence of an aquifer. It becomes apparent that the next generation of geothermal power plants will utilize more of the lower-temperature aquifer resources or the dry resources. For such power plants to be economically competitive, modified or new cycles with higher efficiencies must be used. This paper presents two methods to increase the efficiency of the currently used geothermal cycles. The first uses a binary-flashing system to reduce the overall entropy production, thus, producing more electric power from the resource. The second describes a heat extraction system to be used with dry hot-rock resources.

  5. Indium-tin-oxide thin film transistor biosensors for label-free detection of avian influenza virus H5N1

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Di; Zhuo, Ming [Key Laboratory for Micro-Nano Optoelectronic Devices of Ministry of Education, State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Zhang, Xiaoai [State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing (China); Xu, Cheng; Jiang, Jie [Key Laboratory for Micro-Nano Optoelectronic Devices of Ministry of Education, State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Gao, Fu [State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing (China); Wan, Qing, E-mail: wanqing7686@hotmail.com [Key Laboratory for Micro-Nano Optoelectronic Devices of Ministry of Education, State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Li, Qiuhong, E-mail: liqiuhong2004@hotmail.com [Key Laboratory for Micro-Nano Optoelectronic Devices of Ministry of Education, State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Wang, Taihong, E-mail: thwang@hnu.cn [Key Laboratory for Micro-Nano Optoelectronic Devices of Ministry of Education, State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China)

    2013-04-22

    Highlights: ► A highly selective label-free biosensor is established based on indium-tin-oxide thin-film transistors (ITO TFTs). ► AI H5N1 virus was successfully detected through shift in threshold voltage and field-effect mobility of ITO TFT. ► The ITO TFT is applied in biosensor for the first time and shows good reusability and stability. ► Fabrication of the platform is simple with low cost, which is suitable for mass commercial production. -- Abstract: As continuous outbreak of avian influenza (AI) has become a threat to human health, economic development and social stability, it is urgently necessary to detect the highly pathogenic avian influenza H5N1 virus quickly. In this study, we fabricated indium-tin-oxide thin-film transistors (ITO TFTs) on a glass substrate for the detecting of AI H5N1. The ITO TFT is fabricated by a one-shadow-mask process in which a channel layer can be simultaneously self-assembled between ITO source/drain electrodes during magnetron sputtering deposition. Monoclonal anti-H5N1 antibodies specific for AI H5N1 virus were covalently immobilized on the ITO channel by (3-glycidoxypropyl)trimethoxysilane. The introduction of target AI H5N1 virus affected the electronic properties of the ITO TFT, which caused a change in the resultant threshold voltage (V{sub T}) and field-effect mobility. The changes of I{sub D}–V{sub G} curves were consistent with an n-type field effect transistor behavior affected by nearby negatively charged AI H5N1 viruses. The transistor based sensor demonstrated high selectivity and stability for AI H5N1 virus sensing. The sensor showed linear response to AI H5N1 in the concentrations range from 5 × 10{sup −9} g mL{sup −1} to 5 × 10{sup −6} g mL{sup −1} with a detection limit of 0.8 × 10{sup −10} g mL{sup −1}. Moreover, the ITO TFT biosensors can be repeatedly used through the washing processes. With its excellent electric properties and the potential for mass commercial production, ITO TFTs

  6. The ITO-capped WO{sub 3} nanowires biosensor based on field-effect transistor in label-free protein sensing

    Energy Technology Data Exchange (ETDEWEB)

    Shariati, Mohsen [Sharif University of Technology, Institute for Nanoscience and Nanotechnology, Tehran (Iran, Islamic Republic of)

    2017-05-15

    The fabrication of ITO-capped WO{sub 3} nanowires associated with their bio-sensing properties in field-effect transistor diagnostics basis as a biosensor has been reported. The bio-sensing property for manipulated nanowires elucidated that the grown nanostructures were very sensitive to protein. The ITO-capped WO{sub 3} nanowires biosensor showed an intensive bio-sensing activity against reliable protein. Polylysine strongly charged bio-molecule was applied as model system to demonstrate the implementation of materialized biosensor. The employed sensing mechanism was 'label-free' and depended on bio-molecule's intrinsic charge. For nanowires synthesis, the vapor-liquid-solid mechanism was used. Nanowires were beyond a few hundred nanometers in lengths and around 15-20 nm in diameter, while the globe cap's size on the nanowires was around 15-25 nm. The indium tin oxide (ITO) played as catalyst in nanofabrication for WO{sub 3} nanowires growth and had outstanding role in bio-sensing especially for bio-molecule adherence. In applied electric field presence, the fabricated device showed the great potential to enhance medical diagnostics. (orig.)

  7. Label-Free Sensors Based on Graphene Field-Effect Transistors for the Detection of Human Chorionic Gonadotropin Cancer Risk Biomarker

    Directory of Open Access Journals (Sweden)

    Carrie Haslam

    2018-01-01

    Full Text Available We report on the development of label-free chemical vapour deposition (CVD graphene field effect transistor (GFET immunosensors for the sensitive detection of Human Chorionic Gonadotropin (hCG, a glycoprotein risk biomarker of certain cancers. The GFET sensors were fabricated on Si/SiO2 substrate using photolithography with evaporated chromium and sputtered gold contacts. GFET channels were functionalised with a linker molecule to an immobile anti-hCG antibody on the surface of graphene. The binding reaction of the antibody with varying concentration levels of hCG antigen demonstrated the limit of detection of the GFET sensors to be below 1 pg/mL using four-probe electrical measurements. We also show that annealing can significantly improve the carrier transport properties of GFETs and shift the Dirac point (Fermi level with reduced p-doping in back-gated measurements. The developed GFET biosensors are generic and could find applications in a broad range of medical diagnostics in addition to cancer, such as neurodegenerative (Alzheimer’s and Parkinson’s and cardiovascular disorders.

  8. A label-free and high sensitive aptamer biosensor based on hyperbranched polyester microspheres for thrombin detection

    International Nuclear Information System (INIS)

    Sun, Chong; Han, Qiaorong; Wang, Daoying; Xu, Weimin; Wang, Weijuan; Zhao, Wenbo; Zhou, Min

    2014-01-01

    Highlights: • A label-free thrombin aptamer biosensor applied in whole blood has been developed. • The aptamer biosensor showed a wide detection range and a low detection limit. • The antibiofouling idea utilized for biosensor is significant for diagnostics. - Abstract: In this paper, we have synthesized hyperbranched polyester microspheres with carboxylic acid functional groups (HBPE-CA) and developed a label-free electrochemical aptamer biosensor using thrombin-binding aptamer (TBA) as receptor for the measurement of thrombin in whole blood. The indium tin oxide (ITO) electrode surface modified with HBPE-CA microspheres was grafted with TBA, which has excellent binding affinity and selectivity for thrombin. Binding of the thrombin at the modified ITO electrode surface greatly restrained access of electrons for a redox probe of [Fe(CN) 6 ] 3−/4− . Moreover, the aptamer biosensor could be used for detection of thrombin in whole blood, a wide detection range (10 fM–100 nM) and a detection limit on the order of 0.90 fM were demonstrated. Control experiments were also carried out by using bull serum albumin (BSA) and lysozyme in the absence of thrombin. The good stability and repeatability of this aptamer biosensor were also proved. We expect that this demonstration will lead to the development of highly sensitive label-free sensors based on aptamer with lower cost than current technology. The integration of the technologies, which include anticoagulant, sensor and nanoscience, will bring significant input to high-performance biosensors relevant to diagnostics and therapy of interest for human health

  9. A label-free and high sensitive aptamer biosensor based on hyperbranched polyester microspheres for thrombin detection

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Chong [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014 (China); Han, Qiaorong [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Wang, Daoying; Xu, Weimin [Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014 (China); Wang, Weijuan [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Zhao, Wenbo, E-mail: zhaowenbo@njnu.edu.cn [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Zhou, Min, E-mail: zhouminnju@126.com [Department of Vascular Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008 (China)

    2014-11-19

    Highlights: • A label-free thrombin aptamer biosensor applied in whole blood has been developed. • The aptamer biosensor showed a wide detection range and a low detection limit. • The antibiofouling idea utilized for biosensor is significant for diagnostics. - Abstract: In this paper, we have synthesized hyperbranched polyester microspheres with carboxylic acid functional groups (HBPE-CA) and developed a label-free electrochemical aptamer biosensor using thrombin-binding aptamer (TBA) as receptor for the measurement of thrombin in whole blood. The indium tin oxide (ITO) electrode surface modified with HBPE-CA microspheres was grafted with TBA, which has excellent binding affinity and selectivity for thrombin. Binding of the thrombin at the modified ITO electrode surface greatly restrained access of electrons for a redox probe of [Fe(CN){sub 6}]{sup 3−/4−}. Moreover, the aptamer biosensor could be used for detection of thrombin in whole blood, a wide detection range (10 fM–100 nM) and a detection limit on the order of 0.90 fM were demonstrated. Control experiments were also carried out by using bull serum albumin (BSA) and lysozyme in the absence of thrombin. The good stability and repeatability of this aptamer biosensor were also proved. We expect that this demonstration will lead to the development of highly sensitive label-free sensors based on aptamer with lower cost than current technology. The integration of the technologies, which include anticoagulant, sensor and nanoscience, will bring significant input to high-performance biosensors relevant to diagnostics and therapy of interest for human health.

  10. Label-free image-based detection of drug resistance with optofluidic time-stretch microscopy (Conference Presentation)

    Science.gov (United States)

    Kobayashi, Hirofumi; Lei, Cheng; Mao, Ailin; Jiang, Yiyue; Guo, Baoshan; Ozeki, Yasuyuki; Goda, Keisuke

    2017-02-01

    Acquired drug resistance is a fundamental predicament in cancer therapy. Early detection of drug-resistant cancer cells during or after treatment is expected to benefit patients from unnecessary drug administration and thus play a significant role in the development of a therapeutic strategy. However, the development of an effective method of detecting drug-resistant cancer cells is still in its infancy due to their complex mechanism in drug resistance. To address this problem, we propose and experimentally demonstrate label-free image-based drug resistance detection with optofluidic time-stretch microscopy using leukemia cells (K562 and K562/ADM). By adding adriamycin (ADM) to both K562 and K562/ADM (ADM-resistant K562 cells) cells, both types of cells express unique morphological changes, which are subsequently captured by an optofluidic time-stretch microscope. These unique morphological changes are extracted as image features and are subjected to supervised machine learning for cell classification. We hereby have successfully differentiated K562 and K562/ADM solely with label-free images, which suggests that our technique is capable of detecting drug-resistant cancer cells. Our optofluidic time-stretch microscope consists of a time-stretch microscope with a high spatial resolution of 780 nm at a 1D frame rate of 75 MHz and a microfluidic device that focuses and orders cells. We compare various machine learning algorithms as well as various concentrations of ADM for cell classification. Owing to its unprecedented versatility of using label-free image and its independency from specific molecules, our technique holds great promise for detecting drug resistance of cancer cells for which its underlying mechanism is still unknown or chemical probes are still unavailable.

  11. Quinone-Based Polymers for Label-Free and Reagentless Electrochemical Immunosensors: Application to Proteins, Antibodies and Pesticides Detection

    Directory of Open Access Journals (Sweden)

    Minh-Chau Pham

    2013-01-01

    Full Text Available Polyquinone derivatives are widely recognized in the literature for their remarkable properties, their biocompatibility, simple synthesis, and easy bio-functionalization. We have shown that polyquinones present very stable electroactivity in neutral aqueous medium within the cathodic potential domain avoiding side oxidation of interfering species. Besides, they can act as immobilized redox transducers for probing biomolecular interactions in sensors. Our group has been working on devices based on such modified electrodes with a view to applications for proteins, antibodies and organic pollutants using a reagentless label-free electrochemical immunosensor format. Herein, these developments are briefly reviewed and put into perspective.

  12. Novel charge plasma based dielectric modulated impact ionization MOSFET as a biosensor for label-free detection

    Science.gov (United States)

    Chanda, Manash; Dey, Prithu; De, Swapnadip; Sarkar, Chandan Kumar

    2015-10-01

    In this paper a charge plasma based dielectric modulated impact ionization MOSFET (CP-DIMOSFET) has been proposed for the first time to ease the label free detection of biomolecules. The concept of CP-DIMOSFET is proposed and analyzed on basis of simulated data using SILVACO ATLAS. Low thermal budgeting and thin silicon layer without any dopant implantations make the proposed structure advantageous compared to the existing MOSFET based biosensors. The results show that the proposed device is capable to detect the presence of biomolecules. Simple fabrication schemes, miniaturization, high sensitivity, dominance of dielectric modulation make the proposed biosensor a promising one that could one day revolutionize the healthcare industry.

  13. Assessing Collagen and Elastin Pressure-Dependent Microarchitectures in Live, Human Resistance Arteries by Label-Free Fluorescence Microscopy

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Thorsted, Bjarne; Brewer, Jonathan R.

    2017-01-01

    are performed on live, perfused arteries, however, an alternative approach using standard video-microscopy pressure myography in combination with post-fixation imaging of re-pressurized vessels is discussed. This alternative method provides users with different options for analysis approaches. The inclusion....... In this work, we describe an ex vivo method for passive mechanical testing and simultaneous label-free three-dimensional imaging of the microarchitecture of elastin and collagen in the arterial wall of isolated human resistance arteries. The imaging protocol can be applied to resistance arteries of any species...

  14. The direct conversion of solar energy to electricity

    International Nuclear Information System (INIS)

    1992-01-01

    Half the world's population lives without access to electricity in the rural areas and villages of developing countries. In 1987, world population reached 5 billion and, according to World Bank projections, will increase to over 6 billion in the year 2000 and to over 8 billion in 2025. Such population growth is not uniformly distributed: developed countries have small or negative growth and account for a declining proportion of the world's population. Inasmuch as 95 per cent of the extra inhabitants added each year are in developing countries, rapid population growth in those countries raises serious questions about energy availability for basic human needs and, of course, more broadly about the environment's capacity to support that growth. The present report makes reference to one of the most comprehensively documented conservative scenarios for world energy demand in the year 2020, namely, Energy for a Sustainable World, which assumed that long-term world sustainability must entail constraints on (a) use of natural resources and (b) combustion of fossil fuels resulting in the greenhouse effect. Solar energy is abundant and could become a major source of electricity. Photovoltaics has three particular advantages. It accomplishes sunlight-to-electricity conversion entirely with solid-state electronic components, and with no moving parts required, thereby promising high equipment availability and very low operating and maintenance costs. PV also appears to have very limited environmental impact, with no emissions of the gaseous pollutants associated with fossil-fuel burning and few of the possible local problems associated with some other renewable energy technologies. Finally, the products of photovoltaic technology are modular in construction and can be built up on site in a flexible way, thus minimizing front-end financial risk and investment costs. Figs and tabs

  15. Direct external investment in the Brazilian electric power sector

    International Nuclear Information System (INIS)

    Pinto Junior, Helder Queiroz; Silveira, Joyce Perin; Losekann, Luciano Dias

    1999-01-01

    The central issue of debate was the need to align the energy sector's options and organization with changing global patterns of economic and social development, characterized by the increasing role played by the private sector, greater integration in the world economy, and new economic and social priorities such as efficiency, decentralization, deregulation, and a closer attention to environmental issues. The aim of the work was to identify the foreign investors participants of Brazilian privatization in the electric power sector and to analyze their strategies

  16. High Torque, Direct Drive Electric Motor, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Bear Engineering proposes to advance the development of an innovative high torque, low speed, direct drive motor in order to meet NASA's requirements for such...

  17. High Torque, Direct Drive Electric Motor, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Bear Engineering proposes to develop an innovative high torque, low speed, direct drive motor in order to meet NASA's requirements for such devices. Fundamentally,...

  18. Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor.

    Science.gov (United States)

    Song, Chunmei; Li, Jianwu; Liu, Jinxin; Liu, Qing

    2016-08-15

    A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 10(6)cells/mL while the LOD for semi-quantitative detection could go down to 10(5)cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Performance evaluation and improvement directions for an Indian electric utility

    Energy Technology Data Exchange (ETDEWEB)

    Yadav, Vinod Kumar, E-mail: vinod@gbu.ac.in [Electrical Engineering Department, Gautam Buddha University, Greater Noida 201310 (India); Padhy, N.P.; Gupta, H.O. [Electrical Engineering Department, Indian Institute of Technology, Roorkee 247667 (India)

    2011-11-15

    This study evaluates the performance of 29 Electricity Distribution Divisions (EDDs) of an Indian state - Uttarakhand - deploying Input oriented Data Envelopment Analysis (DEA). The results indicate that the performance of several EDDs is sub-optimal, suggesting the potential for cost reductions and possible reduction in employees number. In the DEA method more than one unit are identified as efficient. Therefore, this study suggests a method for ranking the efficient units by their importance as benchmarks for the inefficient units through benchmark share measure. The bigger the benchmark share, the more important an efficient division is in benchmarking for inefficient ones. Result reveals that plain area divisions are relatively efficient and have higher potential to influence the performance of inefficient EDDs. This study is envisaged to be instrumental to policy makers and managers to increase the operational efficiency of inefficient EDDs and thereby increase the competitiveness in the face of restructuring and liberalization of Indian electricity sector. - Highlights: > Plain area divisions are more effective in integrating resources than hilly divisions. > For prevalent inefficiency two models are developed to varying environmental conditions. > Benchmark share identifies the variable that is influential in increasing the efficiency. > Savings in terms of reduction in O and M cost and number of Employees. > Findings of research work redefine the view point of the utility planners.

  20. Label-free imaging of Drosophila in vivo by coherent anti-Stokes Raman scattering and two-photon excitation autofluorescence microscopy

    Science.gov (United States)

    Chien, Cheng-Hao; Chen, Wei-Wen; Wu, June-Tai; Chang, Ta-Chau

    2011-01-01

    Drosophila is one of the most valuable model organisms for studying genetics and developmental biology. The fat body in Drosophila, which is analogous to the liver and adipose tissue in human, stores lipids that act as an energy source during its development. At the early stages of metamorphosis, the fat body remodeling occurs involving the dissociation of the fat body into individual fat cells. Here we introduce a combination of coherent anti-Stokes Raman scattering (CARS) and two-photon excitation autofluorescence (TPE-F) microscopy to achieve label-free imaging of Drosophila in vivo at larval and pupal stages. The strong CARS signal from lipids allows direct imaging of the larval fat body and pupal fat cells. In addition, the use of TPE-F microscopy allows the observation of other internal organs in the larva and autofluorescent globules in fat cells. During the dissociation of the fat body, the findings of the degradation of lipid droplets and an increase in autofluorescent globules indicate the consumption of lipids and the recruitment of proteins in fat cells. Through in vivo imaging and direct monitoring, CARS microscopy may help elucidate how metamorphosis is regulated and study the lipid metabolism in Drosophila.

  1. Label-Free (XIC) Quantification of Venom Procoagulant and Neurotoxin Expression in Related Australian Elapid Snakes Gives Insight into Venom Toxicity Evolution.

    Science.gov (United States)

    Skejic, Jure; Steer, David L; Dunstan, Nathan; Hodgson, Wayne C

    2015-11-06

    This study demonstrates a direct role of venom protein expression alteration in the evolution of snake venom toxicity. Avian skeletal muscle contractile response to exogenously administered acetylcholine is completely inhibited upon exposure to South Australian and largely preserved following exposure to Queensland eastern brown snake Pseudonaja textilis venom, indicating potent postsynaptic neurotoxicity of the former and lack thereof of the latter venom. Label-free quantitative proteomics reveals extremely large differences in the expression of postsynaptic three-finger α-neurotoxins in these venoms, explaining the difference in the muscle contractile response and suggesting that the type of toxicity induced by venom can be modified by altered expression of venom proteins. Furthermore, the onset of neuromuscular paralysis in the rat phrenic nerve-diaphragm preparation occurs sooner upon exposure to the venom (10 μg/mL) with high expression of α-neurotoxins than the venoms containing predominately presynaptic β-neurotoxins. The study also finds that the onset of rat plasma coagulation is faster following exposure to the venoms with higher expression of venom prothrombin activator subunits. This is the first quantitative proteomic study that uses extracted ion chromatogram peak areas (MS1 XIC) of distinct homologous tryptic peptides to directly show the differences in the expression of venom proteins.

  2. Electrical measurements in the atmosphere and the Ionosphere over an active thunderstorm. II - Direct current electric fields and conductivity

    Science.gov (United States)

    Holzworth, R. H.; Kelley, M. C.; Siefring, C. L.; Hale, L. C.; Mitchell, J. D.

    1985-01-01

    On August 9, 1981, a series of three rockets was launched over an air mass thunderstorm off the eastern seaboard of Virginia while simultaneous stratospheric and ground-based electric field measurements were made. The conductivity was substantially lower at most altitudes than the conductivity profiles used by theoretical models. Direct current electric fields over 80 mV/m were measured as far away as 96 km from the storm in the stratosphere at 23 km altitude. No dc electric fields above 75 km altitude could be identified with the thunderstorm, in agreement with theory. However, vertical current densities over 120 pA/sq m were seen well above the classical 'electrosphere' (at 50 or 60 km). Frequent dc shifts in the electric field following lightning transients were seen by both balloon and rocket payloads. These dc shifts are clearly identifiable with either cloud-to-ground (increases) or intercloud (decreases) lightning flashes.

  3. The morphological and molecular changes of brain cells exposed to direct current electric field stimulation.

    Science.gov (United States)

    Pelletier, Simon J; Lagacé, Marie; St-Amour, Isabelle; Arsenault, Dany; Cisbani, Giulia; Chabrat, Audrey; Fecteau, Shirley; Lévesque, Martin; Cicchetti, Francesca

    2014-12-07

    The application of low-intensity direct current electric fields has been experimentally used in the clinic to treat a number of brain disorders, predominantly using transcranial direct current stimulation approaches. However, the cellular and molecular changes induced by such treatment remain largely unknown. Here, we tested various intensities of direct current electric fields (0, 25, 50, and 100V/m) in a well-controlled in vitro environment in order to investigate the responses of neurons, microglia, and astrocytes to this type of stimulation. This included morphological assessments of the cells, viability, as well as shape and fiber outgrowth relative to the orientation of the direct current electric field. We also undertook enzyme-linked immunosorbent assays and western immunoblotting to identify which molecular pathways were affected by direct current electric fields. In response to direct current electric field, neurons developed an elongated cell body shape with neurite outgrowth that was associated with a significant increase in growth associated protein-43. Fetal midbrain dopaminergic explants grown in a collagen gel matrix also showed a reorientation of their neurites towards the cathode. BV2 microglial cells adopted distinct morphological changes with an increase in cyclooxygenase-2 expression, but these were dependent on whether they had already been activated with lipopolysaccharide. Finally, astrocytes displayed elongated cell bodies with cellular filopodia that were oriented perpendicularly to the direct current electric field. We show that cells of the central nervous system can respond to direct current electric fields both in terms of their morphological shape and molecular expression of certain proteins, and this in turn can help us to begin understand the mechanisms underlying the clinical benefits of direct current electric field. © The Author 2015. Published by Oxford University Press on behalf of CINP.

  4. The development of direct core monitoring in Nuclear Electric plc

    International Nuclear Information System (INIS)

    Curtis, R.F.; Jones, S. Reed, J.; Wickham, A.J.

    1996-01-01

    Monitoring of graphite behaviour in Nuclear Electric Magnox and AGR reactors is necessary to support operating safety cases and to ensure that reactor operation is optimized to sustain the necessary core integrity for the economic life of the reactors. The monitoring programme combines studies for pre-characterized ''installed'' samples with studies on samples trepanned from within the cores and also with studies of core and channel geometry using specially designed equipment. Nuclear Electric has two trepanning machines originally designed for Magnox-reactor work which have been used for a substantial programme over many years. They have recently been upgraded to improve sampling speed, safety and versatility - the last being demonstrated by their adaptation for a recently-won contract associated with decommissioning the Windscale piles. Radiological hazards perceived when the AGR trepanning system was designed resulted in very cumbersome equipment. This has worked well but has been inconvenient in operation. The development of a smaller and improved system for deploying the equipment is now reported. Channel dimension monitoring equipment is discussed in detail with examples of data recovered from both Magnox and AGR cores. A resolution of ± 2 of arc (tilt) and ± 0.01 mm change in diameter in attainable. It is also theoretically possible to establish brick stresses by measuring geometry changes which result from trepanning. Current development work on a revolving scanning laser rangefinder which will enable the measurement of diameters to a resolution of 0.001 mm will also be discussed. This paper also discusses non-destructive techniques for crack detection employing ultrasound or resistance networks, the use of special manipulators to deliver inspection and repair equipment and recent developments to install displacement monitors in peripheral regions of the cores, to aid the understanding of the interaction of the restraint system with the core - the region

  5. 75 FR 77570 - Airworthiness Directives; General Electric Company CF6 Series Turbofan Engines

    Science.gov (United States)

    2010-12-13

    ...) for General Electric (GE) CF6-45/-50 series and CF6-80A series turbofan engines with certain part...-1151; Directorate Identifier 95-ANE-10-AD] RIN 2120-AA64 Airworthiness Directives; General Electric Company CF6 Series Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT. ACTION: Notice of...

  6. A universal and label-free impedimetric biosensing platform for discrimination of single nucleotide substitutions in long nucleic acid strands.

    Science.gov (United States)

    Mills, Dawn M; Martin, Christopher P; Armas, Stephanie M; Calvo-Marzal, Percy; Kolpashchikov, Dmitry M; Chumbimuni-Torres, Karin Y

    2018-06-30

    We report a label-free universal biosensing platform for highly selective detection of long nucleic acid strands. The sensor consists of an electrode-immobilized universal stem-loop (USL) probe and two adaptor strands that form a 4J structure in the presence of a specific DNA/RNA analyte. The sensor was characterized by electrochemical impedance spectroscopy (EIS) using K 3 [Fe(CN) 6 ]/K 4 [Fe(CN) 6 ] redox couple in solution. An increase in charge transfer resistance (R CT ) was observed upon 4J structure formation, the value of which depends on the analyte length. Cyclic voltammetry (CV) was used to further characterize the sensor and monitor the electrochemical reaction in conjunction with thickness measurements of the mixed DNA monolayer obtained using spectroscopic ellipsometry. In addition, the electron transfer was calculated at the electrode/electrolyte interface using a rotating disk electrode. Limits of detection in the femtomolar range were achieved for nucleic acid targets of different lengths (22 nt, 60 nt, 200 nt). The sensor produced only a background signal in the presence of single base mismatched analytes, even in hundred times excess in concentration. This label-free and highly selective biosensing platform is versatile and can be used for universal detection of nucleic acids of varied lengths which could revolutionize point of care diagnostics for applications such as bacterial or cancer screening. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Label-free detection of cellular drug responses by high-throughput bright-field imaging and machine learning.

    Science.gov (United States)

    Kobayashi, Hirofumi; Lei, Cheng; Wu, Yi; Mao, Ailin; Jiang, Yiyue; Guo, Baoshan; Ozeki, Yasuyuki; Goda, Keisuke

    2017-09-29

    In the last decade, high-content screening based on multivariate single-cell imaging has been proven effective in drug discovery to evaluate drug-induced phenotypic variations. Unfortunately, this method inherently requires fluorescent labeling which has several drawbacks. Here we present a label-free method for evaluating cellular drug responses only by high-throughput bright-field imaging with the aid of machine learning algorithms. Specifically, we performed high-throughput bright-field imaging of numerous drug-treated and -untreated cells (N = ~240,000) by optofluidic time-stretch microscopy with high throughput up to 10,000 cells/s and applied machine learning to the cell images to identify their morphological variations which are too subtle for human eyes to detect. Consequently, we achieved a high accuracy of 92% in distinguishing drug-treated and -untreated cells without the need for labeling. Furthermore, we also demonstrated that dose-dependent, drug-induced morphological change from different experiments can be inferred from the classification accuracy of a single classification model. Our work lays the groundwork for label-free drug screening in pharmaceutical science and industry.

  8. Label-free detection of HIV-1 infected cells via integration of optical tweezers and photoluminescence spectroscopy

    Science.gov (United States)

    Lugongolo, Masixole Yvonne; Ombinda-Lemboumba, Saturnin; Noto, Luyanda Lunga; Maaza, Malik; Mthunzi-Kufa, Patience

    2018-02-01

    The human immunodeficiency virus-1 (HIV-1) is currently detected using conventional qualitative and quantitative tests to determine the presence or absence of HIV in blood samples. However, the approach of these tests detects the presence of either viral antibodies or viral RNA that require labelling which may be costly, sophisticated and time consuming. A label-free approach of detecting the presence of HIV is therefore desirable. Of note optical tweezers can be coupled with other technologies including spectroscopy, which also investigates light-matter interactions. For example, coupling of optical tweezers with luminescence spectroscopy techniques has emerged as a powerful tool in biology for micro-manipulation, detection and analysis of individual cells. Integration of optical techniques has enabled studying biological particles in a label-free manner, whilst detecting functional groups and other essential molecules within mixed populations of cells. In the current study, an optical trapping system coupled to luminescence spectroscopy was utilised to detect the presence of HIV infection in TZM-bl cells in vitro. This was performed by infecting TZM-bl cells with the ZM53 HIV-1 pseudovirus, and incubating them for 48 hours prior analysis. The differences between infected and uninfected cells were thereafter displayed as shown by the spectrographs obtained. Combination of these two techniques has a potential in the field of infectious disease diagnostics.

  9. Mapping in vivo target interaction profiles of covalent inhibitors using chemical proteomics with label-free quantification.

    Science.gov (United States)

    van Rooden, Eva J; Florea, Bogdan I; Deng, Hui; Baggelaar, Marc P; van Esbroeck, Annelot C M; Zhou, Juan; Overkleeft, Herman S; van der Stelt, Mario

    2018-04-01

    Activity-based protein profiling (ABPP) has emerged as a valuable chemical proteomics method to guide the therapeutic development of covalent drugs by assessing their on-target engagement and off-target activity. We recently used ABPP to determine the serine hydrolase interaction landscape of the experimental drug BIA 10-2474, thereby providing a potential explanation for the adverse side effects observed with this compound. ABPP allows mapping of protein interaction landscapes of inhibitors in cells, tissues and animal models. Whereas our previous protocol described quantification of proteasome activity using stable-isotope labeling, this protocol describes the procedures for identifying the in vivo selectivity profile of covalent inhibitors with label-free quantitative proteomics. The optimization of our protocol for label-free quantification methods results in high proteome coverage and allows the comparison of multiple biological samples. We demonstrate our protocol by assessing the protein interaction landscape of the diacylglycerol lipase inhibitor DH376 in mouse brain, liver, kidney and testes. The stages of the protocol include tissue lysis, probe incubation, target enrichment, sample preparation, liquid chromatography-mass spectrometry (LC-MS) measurement, data processing and analysis. This approach can be used to study target engagement in a native proteome and to identify potential off targets for the inhibitor under investigation. The entire protocol takes at least 4 d, depending on the number of samples.

  10. Exploration of two-dimensional bio-functionalized phosphorene nanosheets (black phosphorous) for label free haptoglobin electro-immunosensing applications.

    Science.gov (United States)

    Tuteja, Satish K; Neethirajan, Suresh

    2018-02-14

    We report on the development of an antibody-functionalized interface based on electrochemically active liquid-exfoliated two-dimensional phosphorene (Ph) nanosheets-also known as black phosphorous nanosheets-for the label-free electrochemical immunosensing of a haptoglobin (Hp) biomarker, a clinical marker of severe inflammation. The electrodeposition has been achieved over the screen-printed electrode (SPE) using liquid-assisted ultrasonically exfoliated black phosphorus nanosheets. Subsequently, Ph-SPEs bioconjugated with Hp antibodies (Ab), using electrostatic interactions via a poly-L-lysine linker for biointerface development. Electrochemical analysis demonstrates that the Ab-modified Ph-SPEs (Ab@Ph-SPE) exhibit enhanced electroconducting behavior as compared to the pristine electrodes. This Ab-functionalized phosphorene-based electrochemical immunosensor platform has demonstrated remarkable sensitivity and specificity, having a dynamic linear response range from 0.01-10 mg ml -1 for Hp in standard and serum samples with a low detection limit (∼0.011 mg ml -1 ) using the label-free electrochemical technique. The sensor electrodes were also studied with other closely relative interferents to investigate cross reactivity and specificity. This strategy opens up avenues to POC (point-of-care) and on-farm livestock disease monitoring technologies for multiplexed diagnosis in complex biological samples such as serum. The technique is simple in fabrication and provides an analytical response in less than 60 s.

  11. Exploration of 2-Dimensional Bio-functionalized Phosphorene Nanosheets (Black Phosphorous) for Label free Haptoglobin Electro-immunosensing Applications.

    Science.gov (United States)

    Tuteja, Satish; Neethirajan, Suresh

    2018-01-29

    We report on the development of an antibody-functionalized interface based on electrochemically active liquid-exfoliated two-dimensional phosphorene (Ph) nanosheets-also known as black phosphorous nanosheets-for the label-free electrochemical immunosensing of a haptoglobin (Hp) biomarker, a clinical marker of severe inflammation. The electrodeposition has been achieved over the screen-printed electrode (SPE) using liquid-assisted ultrasonically exfoliated black phosphorus nanosheets. Subsequently, Ph-SPEs bioconjugated with Hp antibodies (Ab), using electrostatic interactions via a poly-L-lysine (PLL) linker for biointerface development. Electrochemical analysis demonstrates that the Ab-modified Ph-SPEs (Ab@Ph-SPE) exhibit enhanced electroconducting behavior as compared to the pristine electrodes. This Ab-functionalized phosphorene-based electrochemical immunosensor platform has demonstrated remarkable sensitivity and specificity, having a dynamic linear response range from 0.01 mg/mL to 10 mg/mL for Hp in standard and serum samples with a low detection limit (∼0.011 mg/mL) using the label-free electrochemical technique. The sensor electrodes were also studied with other closely relative interferents to investigate cross reactivity and specificity. This strategy opens up avenues to POC (point-of-care) and on-farm livestock disease monitoring technologies for multiplexed diagnosis in complex biological samples such as serum. The technique is simple in fabrication and provides an analytical response in less than 60 seconds. © 2018 IOP Publishing Ltd.

  12. Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC-MS.

    Science.gov (United States)

    Pando-Robles, Victoria; Oses-Prieto, Juan A; Rodríguez-Gandarilla, Myriam; Meneses-Romero, Erika; Burlingame, Alma L; Batista, Cesar V F

    2014-12-05

    Dengue is an important and growing public health problem worldwide with an estimated 100million new clinical cases annually. Currently, no licensed drug or vaccine is available. During natural infection in humans, liver cells constitute one of the main targets of dengue virus (DENV) replication. However, a clear understanding of dengue pathogenesis remains elusive. In order to gain a better reading of the cross talk between virus and host cell proteins, we used a proteomics approach to analyze the host response to DENV infection in a hepatic cell line Huh-7. Differences in proteome expression were assayed 24h post-infection using label-free LC-MS. Quantitative analysis revealed 155 differentially expressed proteins, 64 of which were up-regulated and 91 down-regulated. These results reveal an important decrease in the expression of enzymes involved in the glycolytic pathway, citrate cycle, and pyruvate metabolism. This study provides large-scale quantitative information regarding protein expression in the early stages of infection that should be useful for better compression of the pathogenesis of dengue. Dengue infection involves alterations in the homeostasis of the host cell. Defining the interactions between virus and cell proteins should provide a better understanding of how viruses propagate and cause disease. Here, we present for the first time the proteomic analysis of hepatocytes (Huh-7 cells) infected with DENV-2 by label-free LC-MS. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Description of an Advantageous Optical Label-Free Biosensing Interferometric Read-Out Method to Measure Biological Species

    Directory of Open Access Journals (Sweden)

    Miguel Holgado

    2014-02-01

    Full Text Available In this article we report a new, simple, and reliable optical read-out detection method able to assess Rotavirus present in human sera as well as in the viral pollution sources. It is based on the interference of two interferometers used as biophotonic transducers. The method significantly improves the optical label-free biosensing response measuring both, the concentration of the AgR and its corresponding size. Two different immunoassays were carried out: Bovine Serum Albumin (BSA, and the recognition by its antibody (anti-BSA; and Rotavirus (AgR and the recognition by its antibody (anti-AgR. In the cases studied, and using as model interferometer a simple Fabry-Perot transducer, we demonstrate a biosensing enhancement of two orders of magnitude in the Limit of Detection (LoD. In fact, this read-out optical method may have significant implications to enhance other optical label-free photonic transducers reported in the scientific literature.

  14. Trifunctional fluorescent unnatural nucleoside: Label free detection of T-T/C-C base mismatches, abasic site and bulge DNA.

    Science.gov (United States)

    Bag, Subhendu Sekhar; Pradhan, Manoj Kumar; Talukdar, Sangita

    2017-08-01

    The detection and targeting of both the mismatched and abasic DNA is highly important which would ultimately help in designing new diagnostics and chemotherapeutics. Furthermore, sensing and targeting the bulge sequence with a fluorescent probe would be useful to study the role of bulges in nucleic acid function or could have significant therapeutic potential. Thus, detection of specific bulges by small fluorescent molecules is an attractive research area since the past several years. Many attempts have been made to prepare such compounds. We report herein a label free strategy for the detection of pyrimidine base mismatches (T/T and C/C), sensing of abasic site, and pyrimidine base bulge DNA using an unnatural tetrazolylpyrene nucleoside ( TPy B Do ) as a bare fluorescent probe. The H-bonding/hydrophobic force mediated interactions allow the sensing of all three deformed DNA via an enhancement of fluorescence signal using our simple "Just-Mix and Read" strategy. The binding of the probe to all the three deformed DNA duplexes is accompanied by an increase in the thermal melting stability of the deformed DNAs. That the probe binds efficiently to the minor groove near the deformed site was evident from spectroscopic studies. All the spectral evidences open up a multitude of possibilities for using our probe, tetrazolylpyrene nucleoside, as an efficient fluorescent light-up bio-probe for label free DNA detection. Copyright © 2017. Published by Elsevier B.V.

  15. Label-free hyperspectral imaging and quantification methods for surgical margin assessment of tissue specimens of cancer patients.

    Science.gov (United States)

    Fei, Baowei; Guolan Lu; Halicek, Martin T; Wang, Xu; Zhang, Hongzheng; Little, James V; Magliocca, Kelly R; Patel, Mihir; Griffith, Christopher C; El-Deiry, Mark W; Chen, Amy Y

    2017-07-01

    Hyperspectral imaging (HSI) is a relatively new modality in medicine and can have many potential applications. In this study, we developed label-free hyperspectral imaging for tumor margin assessment. HSI data, hypercube (x,y,λ), consists of a series of images of the same field of view that are acquired at different wavelengths. Every pixel in the hypercube has an optical spectrum. We collected surgical tissue specimens from 16 human subjects who underwent head and neck (H&N) cancer surgery. We acquired both HSI, autofluorescence images, and fluorescence images with 2-NBDG and proflavine from the specimens. Digitized histologic slides were examined by an H&N pathologist. We developed image preprocessing and classification methods for HSI data and differentiate cancer from benign tissue. The hyperspectral imaging and classification method was able to distinguish between cancer and normal tissue from oral cavity with an average accuracy of 90±8%, sensitivity of 89±9%, and specificity of 91±6%. This study suggests that label-free hyperspectral imaging has great potential for surgical margin assessment in tissue specimens of H&N cancer patients. Further development of the imaging technology and quantification methods is warranted for its application in image-guided surgery.

  16. A Label-Free Electrochemical Impedance Cytosensor Based on Specific Peptide-Fused Phage Selected from Landscape Phage Library

    Science.gov (United States)

    Han, Lei; Liu, Pei; Petrenko, Valery A.; Liu, Aihua

    2016-02-01

    One of the major challenges in the design of biosensors for cancer diagnosis is to introduce a low-cost and selective probe that can recognize cancer cells. In this paper, we combined the phage display technology and electrochemical impedance spectroscopy (EIS) to develop a label-free cytosensor for the detection of cancer cells, without complicated purification of recognition elements. Fabrication steps of the cytosensing interface were monitored by EIS. Due to the high specificity of the displayed octapeptides and avidity effect of their multicopy display on the phage scaffold, good biocompatibility of recombinant phage, the fibrous nanostructure of phage, and the inherent merits of EIS technology, the proposed cytosensor demonstrated a wide linear range (2.0 × 102 - 2.0 × 108 cells mL-1), a low limit of detection (79 cells mL-1, S/N = 3), high specificity, good inter-and intra-assay reproducibility and satisfactory storage stability. This novel cytosensor designing strategy will open a new prospect for rapid and label-free electrochemical platform for tumor diagnosis.

  17. Label-Free Detection of Rare Cell in Human Blood Using Gold Nano Slit Surface Plasmon Resonance

    Directory of Open Access Journals (Sweden)

    Mansoureh Z. Mousavi

    2015-03-01

    Full Text Available Label-free detection of rare cells in biological samples is an important and highly demanded task for clinical applications and various fields of research, such as detection of circulating tumor cells for cancer therapy and stem cells studies. Surface Plasmon Resonance (SPR as a label-free method is a promising technology for detection of rare cells for diagnosis or research applications. Short detection depth of SPR (400 nm provides a sensitive method with minimum interference of non-targets in the biological samples. In this work, we developed a novel microfluidic chip integrated with gold nanoslit SPR platform for highly efficient immunomagnetic capturing and detection of rare cells in human blood. Our method offers simple yet efficient detection of target cells with high purity. The approach for detection consists of two steps. Target cells are firs captured on functionalized magnetic nanoparticles (MNPs with specific antibody I. The suspension containing the captured cells (MNPs-cells is then introduced into a microfluidic chip integrated with a gold nanoslit film. MNPs-cells bind with the second specific antibody immobilized on the surface of the gold nanoslit and are therefore captured on the sensor active area. The cell binding on the gold nanoslit was monitored by the wavelength shift of the SPR spectrum generated by the gold nanoslits.

  18. A novel label-free electrochemical immunosensor based on functionalized nitrogen-doped graphene quantum dots for carcinoembryonic antigen detection.

    Science.gov (United States)

    Yang, Yuying; Liu, Qing; Liu, Yan; Cui, Jianjian; Liu, Hui; Wang, Ping; Li, Yueyun; Chen, Lei; Zhao, Zengdian; Dong, Yunhui

    2017-04-15

    A novel and ultrasensitive label-free electrochemical immunosensor was fabricated for quantitative detection of carcino-embryonic antigen (CEA). The nitrogen-doped graphene quantum dots (N-GQDs) supported PtPd bimetallic nanoparticles (PtPd/N-GQDs) were synthesized by a simple and green hydrothermal procedure. Subsequently, PtPd/N-GQDs functionalized Au nanoparticles (PtPd/N-GQDs@Au) were prepared successfully via a self-assembly approach. Because of the synergetic effect present in PtPd/N-GQDs@Au, this novel nanocomposites has shown excellent electrocatalytic activity towards hydrogen peroxide (H 2 O 2 ) reduction. Featuring good biocompatibility, excellent conductivity and large surface area, PtPd/N-GQDs@Au was applied as transducing materials to efficiently conjugate capture antibodies and amplify electrochemical signal. Under the optimal conditions, the proposed immunosensor was used for the detection of CEA with wide dynamic range in the range from 5 fg/mL to 50ng/mL with a low detection limit of 2fg/mL (S/N=3). Furthermore, this label-free immunosensor possesses high sensitivity, special selectivity and long-term stability, which shows promising application in bioassay analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Multimodal label-free ex vivo imaging using a dual-wavelength microscope with axial chromatic aberration compensation.

    Science.gov (United States)

    Filippi, Andrea; Dal Sasso, Eleonora; Iop, Laura; Armani, Andrea; Gintoli, Michele; Sandri, Marco; Gerosa, Gino; Romanato, Filippo; Borile, Giulia

    2018-03-01

    Label-free microscopy is a very powerful technique that can be applied to study samples with no need for exogenous fluorescent probes, keeping the main benefits of multiphoton microscopy, such as longer penetration depths and intrinsic optical sectioning while enabling serial multitechniques examinations on the same specimen. Among the many label-free microscopy methods, harmonic generation (HG) is one of the most intriguing methods due to its generally low photo-toxicity and relative ease of implementation. Today, HG and common two-photon microscopy (TPM) are well-established techniques, and are routinely used in several research fields. However, they require a significant amount of fine-tuning to be fully exploited, making them quite difficult to perform in parallel. Here, we present our designed multimodal microscope, capable of performing simultaneously TPM and HG without any kind of compromise thanks to two, separate, individually optimized laser sources with axial chromatic aberration compensation. We also apply our setup to the examination of a plethora of ex vivo samples to prove its capabilities and the significant advantages of a multimodal approach. (2018) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

  20. A label-free colorimetric sensor for Pb2+ detection based on the acceleration of gold leaching by graphene oxide.

    Science.gov (United States)

    Shi, Xinhao; Gu, Wei; Zhang, Cuiling; Zhao, Longyun; Peng, Weidong; Xian, Yuezhong

    2015-03-14

    In this work, we developed a novel, label-free, colorimetric sensor for Pb(2+) detection based on the acceleration of gold leaching by graphene oxide (GO) at room temperature. Gold nanoparticles (AuNPs) can be dissolved in a thiosulfate (S2O3(2-)) aqueous environment in the presence of oxygen; however, the leaching rate is very slow due to the high activation energy (27.99 kJ mol(-1)). In order to enhance the reaction rate, some accelerators should be added. In comparison with the traditional accelerators (metal ions or middle ligands), we found that GO could efficiently accelerate the gold leaching reaction. Kinetic data demonstrate that the dissolution rate of gold in the Pb(2+)-S2O3(2-)-GO system is 5 times faster than that without GO at room temperature. In addition, the effects of surface modification and the nanoparticle size on the etching of AuNPs were investigated. Based on the GO-accelerated concentration-dependent colour changes of AuNPs, a colorimetric sensor for Pb(2+) detection was developed with a linear range from 0.1 to 20 μM and the limit of detection (LOD) was evaluated to be 0.05 μM. This colorimetric assay is simple, low-cost, label-free, and has numerous potential applications in the field of environmental chemistry.

  1. Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

    Directory of Open Access Journals (Sweden)

    Cai-Xia Zhuo

    2016-11-01

    Full Text Available Trypsin is important during the regulation of pancreatic exocrine function. The detection of trypsin activity is currently limited because of the need for the substrate to be labeled with a fluorescent tag. A label-free fluorescent method has been developed to monitor trypsin activity. The designed peptide probe consists of six arginine molecules and a cysteine terminus and can be conjugated to DNA-stabilized silver nanoclusters (DNA-AgNCs by Ag-S bonding to enhance fluorescence. The peptide probe can also be adsorbed to the surface of graphene oxide (GO, thus resulting in the fluorescence quenching of DNA-AgNCs-peptide conjugate because of Förster resonance energy transfer. Once trypsin had degraded the peptide probe into amino acid residues, the DNA-AgNCs were released from the surface of GO, and the enhanced fluorescence of DNA-AgNCs was restored. Trypsin can be determined with a linear range of 0.0–50.0 ng/mL with a concentration as low as 1 ng/mL. This label-free method is simple and sensitive and has been successfully used for the determination of trypsin in serum. The method can also be modified to detect other proteases.

  2. Label-free chemical imaging of live Euglena gracilis by high-speed SRS spectral microscopy (Conference Presentation)

    Science.gov (United States)

    Wakisaka, Yoshifumi; Suzuki, Yuta; Tokunaga, Kyoya; Hirose, Misa; Domon, Ryota; Akaho, Rina; Kuroshima, Mai; Tsumura, Norimichi; Shimobaba, Tomoyoshi; Iwata, Osamu; Suzuki, Kengo; Nakashima, Ayaka; Goda, Keisuke; Ozeki, Yasuyuki

    2016-03-01

    Microbes, especially microalgae, have recently been of great interest for developing novel biofuels, drugs, and biomaterials. Imaging-based screening of live cells can provide high selectivity and is attractive for efficient bio-production from microalgae. Although conventional cellular screening techniques use cell labeling, labeling of microbes is still under development and can interfere with their cellular functions. Furthermore, since live microbes move and change their shapes rapidly, a high-speed imaging technique is required to suppress motion artifacts. Stimulated Raman scattering (SRS) microscopy allows for label-free and high-speed spectral imaging, which helps us visualize chemical components inside biological cells and tissues. Here we demonstrate high-speed SRS imaging, with temporal resolution of 0.14 seconds, of intracellular distributions of lipid, polysaccharide, and chlorophyll concentrations in rapidly moving Euglena gracilis, a unicellular phytoflagellate. Furthermore, we show that our method allows us to analyze the amount of chemical components inside each living cell. Our results indicate that SRS imaging may be applied to label-free screening of living microbes based on chemical information.

  3. Comparison of one- and two-dimensional liquid chromatography approaches in the label-free quantitative analysis of Methylocella silvestris.

    Science.gov (United States)

    Patel, Nisha A; Crombie, Andrew; Slade, Susan E; Thalassinos, Konstantinos; Hughes, Chris; Connolly, Joanne B; Langridge, James; Murrell, J Colin; Scrivens, James H

    2012-09-07

    The proteome of the bacterium Methylocella silvestris has been characterized using reversed phase ultra high pressure liquid chromatography (UPLC) and two-dimensional reversed phase (high pH)-reversed phase (low pH) UPLC prior to mass spectrometric analysis. Variations in protein expression levels were identified with the aid of label-free quantification in a study of soluble protein extracts from the organism grown using methane, succinate, or propane as a substrate. The number of first dimensional fractionation steps has been varied for 2D analyses, and the impact on data throughput and quality has been demonstrated. Comparisons have been made regarding required experimental considerations including total loading of biological samples required, instrument time, and resulting data file sizes. The data obtained have been evaluated with respect to number of protein identifications, confidence of assignments, sequence coverage, relative levels of proteins, and dynamic range. Good qualitative and quantitative agreement was observed between the different approaches, and the potential benefits and limitations of the reversed phase-reversed phase UPLC technique in label-free analysis are discussed. A preliminary screen of the protein regulation data has also been performed, providing evidence for a possible propane assimilation route.

  4. Exploration of two-dimensional bio-functionalized phosphorene nanosheets (black phosphorous) for label free haptoglobin electro-immunosensing applications

    Science.gov (United States)

    Tuteja, Satish K.; Neethirajan, Suresh

    2018-04-01

    We report on the development of an antibody-functionalized interface based on electrochemically active liquid-exfoliated two-dimensional phosphorene (Ph) nanosheets—also known as black phosphorous nanosheets—for the label-free electrochemical immunosensing of a haptoglobin (Hp) biomarker, a clinical marker of severe inflammation. The electrodeposition has been achieved over the screen-printed electrode (SPE) using liquid-assisted ultrasonically exfoliated black phosphorus nanosheets. Subsequently, Ph-SPEs bioconjugated with Hp antibodies (Ab), using electrostatic interactions via a poly-L-lysine linker for biointerface development. Electrochemical analysis demonstrates that the Ab-modified Ph-SPEs (Ab@Ph-SPE) exhibit enhanced electroconducting behavior as compared to the pristine electrodes. This Ab-functionalized phosphorene-based electrochemical immunosensor platform has demonstrated remarkable sensitivity and specificity, having a dynamic linear response range from 0.01–10 mg ml‑1 for Hp in standard and serum samples with a low detection limit (∼0.011 mg ml‑1) using the label-free electrochemical technique. The sensor electrodes were also studied with other closely relative interferents to investigate cross reactivity and specificity. This strategy opens up avenues to POC (point-of-care) and on-farm livestock disease monitoring technologies for multiplexed diagnosis in complex biological samples such as serum. The technique is simple in fabrication and provides an analytical response in less than 60 s.

  5. A direct and at nanometer scale study of electrical charge distribution on membranes of alive cells

    Directory of Open Access Journals (Sweden)

    Marlière Christian

    2016-01-01

    Full Text Available In this paper is presented an innovative method to map in-vivo and at nanometer scale the electrical charge distribution on membranes of alive cells. It relies on a new atomic force microscopy (AFM mode based on an electro-mechanical coupling effect. Furthermore, an additional electrical signal detected by both the deflection of the AFM cantilever and simultaneous direct current measurements was detected at low scanning rates. It was attributed to the detection of the current stemming from ionic channels. It opens a new way to directly investigate in situ biological electrical surface processes involved in bacterial adhesion, biofilm formation, microbial fuel cells, etc.

  6. Proposal for a directive for the promotion of electricity based on renewable energy sources

    International Nuclear Information System (INIS)

    2000-04-01

    The amended ''Directives concerning common rules for the internal markets in electricity and natural gas'', adopted in June 2003, organizes the future framework of electricity and gas, making all European consumers eligible, from 2004 onwards and at the latest by 2007 for the domestic sector, as well as integrating some components related to general interest services. Energie-Cites gives in this document its opinion and its expectations concerning this proposal for a directive for the promotion of electricity based on renewable energy sources. (A.L.B.)

  7. Direct observation of electric field induced pattern formation and particle aggregation in ferrofluids

    Energy Technology Data Exchange (ETDEWEB)

    Rajnak, Michal; Kopcansky, Peter; Timko, Milan [Institute of Experimental Physics SAS, Watsonova 47, 04001 Košice (Slovakia); Petrenko, Viktor I. [Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Kyiv Taras Shevchenko National University, Volodymyrska Street 64, Kyiv 01033 (Ukraine); Avdeev, Mikhail V. [Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Ivankov, Olexandr I. [Joint Institute for Nuclear Research, Joliot-Curie 6, 141980 Dubna, Moscow Region (Russian Federation); Kyiv Taras Shevchenko National University, Volodymyrska Street 64, Kyiv 01033 (Ukraine); Moscow Institute of Physics and Technology, Institutskiy per. 9, Dolgoprudniy 141700 (Russian Federation); Feoktystov, Artem [Jülich Centre for Neutron Science (JCNS) at Heinz Maier-Leibnitz Zentrum (MLZ), Forschungszentrum Jülich GmbH, Lichtenbergstr. 1, 85747 Garching (Germany); Dolnik, Bystrik; Kurimsky, Juraj [Faculty of Electrical Engineering and Informatics, Technical University of Košice, Letná 9, 04200 Košice (Slovakia)

    2015-08-17

    Ferrofluids typically respond to magnetic fields and can be manipulated by external magnetic fields. Here, we report on formation of visually observable patterns in a diluted low-polarity ferrofluid exposed to external electric fields. This presents a specific type of ferrofluid structure driven by a combined effect of electrohydrodynamics and electrical body forces. The free charge and permittivity variation are considered to play a key role in the observed phenomenon. The corresponding changes in the ferrofluid structure have been found at nanoscale as well. By small-angle neutron scattering (SANS), we show that the magnetic nanoparticles aggregate in direct current (dc) electric field with a strong dependence on the field intensity. The anisotropic aggregates preferably orient in the direction of the applied electric field. Conducting SANS experiments with alternating current (ac) electric fields of various frequencies, we found a critical frequency triggering the aggregation process. Our experimental study could open future applications of ferrofluids based on insulating liquids.

  8. Direct observation of electric field induced pattern formation and particle aggregation in ferrofluids

    Science.gov (United States)

    Rajnak, Michal; Petrenko, Viktor I.; Avdeev, Mikhail V.; Ivankov, Olexandr I.; Feoktystov, Artem; Dolnik, Bystrik; Kurimsky, Juraj; Kopcansky, Peter; Timko, Milan

    2015-08-01

    Ferrofluids typically respond to magnetic fields and can be manipulated by external magnetic fields. Here, we report on formation of visually observable patterns in a diluted low-polarity ferrofluid exposed to external electric fields. This presents a specific type of ferrofluid structure driven by a combined effect of electrohydrodynamics and electrical body forces. The free charge and permittivity variation are considered to play a key role in the observed phenomenon. The corresponding changes in the ferrofluid structure have been found at nanoscale as well. By small-angle neutron scattering (SANS), we show that the magnetic nanoparticles aggregate in direct current (dc) electric field with a strong dependence on the field intensity. The anisotropic aggregates preferably orient in the direction of the applied electric field. Conducting SANS experiments with alternating current (ac) electric fields of various frequencies, we found a critical frequency triggering the aggregation process. Our experimental study could open future applications of ferrofluids based on insulating liquids.

  9. Direct comparison of hydraulic tortuosity and electric tortuosity based on finite element analysis

    Directory of Open Access Journals (Sweden)

    H. Saomoto

    2015-08-01

    Full Text Available Tortuosity is one of the key parameters to characterize the transport properties of porous media. There are many models for tortuosity estimation based on some definitions: geometric, hydraulic, electric, and diffusive definitions. However, relationships among those tortuosities remain unclear due to the lack of direct comparison on the same porous media. Here we focus on hydraulic and electric tortuosities and have conducted a series of finite element simulations with the Navier–Stokes equation and the equation for electric current to directly compare tortuosities. The results revealed that: (1 on average, hydraulic tortuosity is 15% greater than that of the electric one; (2 the proposed model based on the van Genuchten-type function successfully approximates both hydraulic and electric tortuosities; (3 tortuosities obtained from the porous media packed with circular particles and square particles show quantitatively similar trends.

  10. Identification of Host Defense-Related Proteins Using Label-Free Quantitative Proteomic Analysis of Milk Whey from Cows with Staphylococcus aureus Subclinical Mastitis

    Directory of Open Access Journals (Sweden)

    Shaimaa Abdelmegid

    2017-12-01

    Full Text Available Staphylococcus aureus is the most common contagious pathogen associated with bovine subclinical mastitis. Current diagnosis of S. aureus mastitis is based on bacteriological culture of milk samples and somatic cell counts, which lack either sensitivity or specificity. Identification of milk proteins that contribute to host defense and their variable responses to pathogenic stimuli would enable the characterization of putative biomarkers of subclinical mastitis. To accomplish this, milk whey samples from healthy and mastitic dairy cows were analyzed using a label-free quantitative proteomics approach. In total, 90 proteins were identified, of which 25 showed significant differential abundance between healthy and mastitic samples. In silico functional analyses indicated the involvement of the differentially abundant proteins in biological mechanisms and signaling pathways related to host defense including pathogen-recognition, direct antimicrobial function, and the acute-phase response. This proteomics and bioinformatics analysis not only facilitates the identification of putative biomarkers of S. aureus subclinical mastitis but also recapitulates previous findings demonstrating the abundance of host defense proteins in intramammary infection. All mass spectrometry data are available via ProteomeXchange with identifier PXD007516.

  11. Enzyme-free and label-free ultrasensitive electrochemical detection of DNA and adenosine triphosphate by dendritic DNA concatamer-based signal amplification.

    Science.gov (United States)

    Liu, Shufeng; Lin, Ying; Liu, Tao; Cheng, Chuanbin; Wei, Wenji; Wang, Li; Li, Feng

    2014-06-15

    Hybridization chain reaction (HCR) strategy has been well developed for the fabrication of various biosensing platforms for signal amplification. Herein, a novel enzyme-free and label-free ultrasensitive electrochemical DNA biosensing platform for the detection of target DNA and adenosine triphosphate (ATP) was firstly proposed, in which three auxiliary DNA probes were ingeniously designed to construct the dendritic DNA concatamer via HCR strategy and used as hexaammineruthenium(III) chloride (RuHex) carrier for signal amplification. With the developed dendritic DNA concatamer-based signal amplification strategy, the DNA biosensor could achieve an ultrasensitive electrochemical detection of DNA and ATP with a superior detection limit as low as 5 aM and 20 fM, respectively, and also demonstrate a high selectivity for DNA and ATP detection. The currently proposed dendritic DNA concatamer opens a promising direction to construct ultrasensitive DNA biosensing platform for biomolecular detection in bioanalysis and clinical biomedicine, which offers the distinct advantages of simplicity and cost efficiency owing to no need of any kind of enzyme, chemical modification or labeling. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Mapping the intracellular distribution of carbon nanotubes after targeted delivery to carcinoma cells using confocal Raman imaging as a label-free technique

    International Nuclear Information System (INIS)

    Lamprecht, C; Unterauer, B; Plochberger, B; Brameshuber, M; Hinterdorfer, P; Ebner, A; Gierlinger, N; Hild, S; Heister, E

    2012-01-01

    The uptake of carbon nanotubes (CNTs) by mammalian cells and their distribution within cells is being widely studied in recent years due to their increasing use for biomedical purposes. The two main imaging techniques used are confocal fluorescence microscopy and transmission electron microscopy (TEM). The former, however, requires labeling of the CNTs with fluorescent dyes, while the latter is a work-intensive technique that is unsuitable for in situ bio-imaging. Raman spectroscopy, on the other hand, presents a direct, straightforward and label-free alternative. Confocal Raman microscopy can be used to image the CNTs inside cells, exploiting the strong Raman signal connected to different vibrational modes of the nanotubes. In addition, cellular components, such as the endoplasmic reticulum and the nucleus, can be mapped. We first validate our method by showing that only when using the CNTs’ G band for intracellular mapping accurate results can be obtained, as mapping of the radial breathing mode (RBM) only shows a small fraction of CNTs. We then take a closer look at the exact localization of the nanotubes inside cells after folate receptor-mediated endocytosis and show that, after 8-10 h incubation, the majority of CNTs are localized around the nucleus. In summary, Raman imaging has enormous potential for imaging CNTs inside cells, which is yet to be fully realized. (paper)

  13. Ultrasensitive in situ label-free DNA detection using a GaN nanowire-based extended-gate field-effect-transistor sensor.

    Science.gov (United States)

    Chen, Chin-Pei; Ganguly, Abhijit; Lu, Ching-Ying; Chen, Ting-Yu; Kuo, Chun-Chiang; Chen, Reui-San; Tu, Wen-Hsun; Fischer, Wolfgang B; Chen, Kuei-Hsien; Chen, Li-Chyong

    2011-03-15

    In this study, we have successfully demonstrated that a GaN nanowire (GaNNW) based extended-gate field-effect-transistor (EGFET) biosensor is capable of specific DNA sequence identification under label-free in situ conditions. Our approach shows excellent integration of the wide bandgap semiconducting nature of GaN, surface-sensitivity of the NW-structure, and high transducing performance of the EGFET-design. The simple sensor-architecture, by direct assembly of as-synthesized GaNNWs with a commercial FET device, can achieve an ultrahigh detection limit below attomolar level concentrations: about 3 orders of magnitude higher in resolution than that of other FET-based DNA-sensors. Comparative in situ studies on mismatches ("hotspot" mutations related to human p53 tumor-suppressor gene) and complementary targets reveal excellent selectivity and specificity of the sensor, even in the presence of noncomplementary DNA strands, suggesting the potential pragmatic application in complex clinical samples. In comparison with GaN thin film, NW-based EGFET exhibits excellent performance with about 2 orders higher sensitivity, over a wide detection range, 10(-19)-10(-6) M, reaching about a 6-orders lower detection limit. Investigations illustrate the unique and distinguished feature of nanomaterials. Detailed studies indicate a positive effect of energy band alignment at the biomaterials-semiconductor hybrid interface influencing the effective capacitance and carrier-mobility of the system.

  14. Label-free impedimetric aptasensor for detection of femtomole level acetamiprid using gold nanoparticles decorated multiwalled carbon nanotube-reduced graphene oxide nanoribbon composites.

    Science.gov (United States)

    Fei, Airong; Liu, Qian; Huan, Juan; Qian, Jing; Dong, Xiaoya; Qiu, Baijing; Mao, Hanping; Wang, Kun

    2015-08-15

    Gold nanoparticles (Au NPs) decorated multiwalled carbon nanotube-reduced graphene oxide nanoribbon (Au/MWCNT-rGONR) composites were synthesized by a one-pot reaction. By employing the resulting Au/MWCNT-rGONR composites as the support for aptamer immobilization, we developed an ultrasensitive label-free electrochemical impedimetric aptasensor for acetamiprid detection, which was based on that the variation of electron transfer resistance was relevant to the formation of acetamiprid-aptamer complex at the modified electrode surface. Compared with pure Au NPs and MWCNT-rGONR, the Au/MWCNT-rGONR composites modified electrode was the most sensitive aptasensing platform for the determination of acetamiprid. The proposed aptasensor displayed a linear response for acetamiprid in the range from 5×10(-14) M to 1×10(-5) M with an extremely low detection limit of 1.7×10(-14) M (S/N=3). In addition, this impedimetric aptasensor possessed great advantages including the simple operation process, low-cost, selectivity and sensitivity, which provided a promising model for the aptamer-based detection with a direct impedimetric method. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Proximity hybridization triggered hemin/G-quadruplex formation for construction a label-free and signal-on electrochemical DNA sensor.

    Science.gov (United States)

    Gao, Fenglei; Fan, Taotao; Wu, Jing; Liu, Sha; Du, Yan; Yao, Yao; Zhou, Fuyi; Zhang, Yu; Liao, Xianjiu; Geng, Deqin

    2017-10-15

    We describe a novel label-free and signal-on electrochemical DNA sensing platform via proximity hybridization triggered hemin/G-quadruplex formation based on the direct electron transfer of hemin. The thiolated modified G-DNA1 was first immobilized onto the Au electrode surface. In the presence of target DNA, Y-junction-structure ternary complex can be formed to trigger the proximity assembly of G-DNA1, hemin, and G-DNA2, which leads to the formation of hemin/G-quadruplex for generation an amplified electrochemical signal by differential pulse voltammetry. The observed signal gain was sufficient to achieve a demonstrated detection limit of 54 fM, with a wide linear dynamic range from 10 -13 to 10 -9 M and discriminated mismatched DNA from perfect matched target DNA with a high selectivity. Benefiting from the one step proximity dependent hemin/G-quadruplex formation, the assay method is extremely simple and can be carried out within 40min. The advantages of free of any label conjugation step, and versatility make it a promising candidate for point-of-care testing and commercial application. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Thermal and electrical energy yield analysis of a directly water cooled photovoltaic module

    Directory of Open Access Journals (Sweden)

    Mtunzi Busiso

    2016-01-01

    Full Text Available Electrical energy of photovoltaic modules drops by 0.5% for each degree increase in temperature. Direct water cooling of photovoltaic modules was found to give improved electrical and thermal yield. A prototype was put in place to analyse the field data for a period of a year. The results showed an initial high performance ratio and electrical power output. The monthly energy saving efficiency of the directly water cooled module was found to be approximately 61%. The solar utilisation of the naturally cooled photovoltaic module was found to be 8.79% and for the directly water cooled module its solar utilisation was 47.93%. Implementation of such systems on households may reduce the load from the utility company, bring about huge savings on electricity bills and help in reducing carbon emissions.

  17. Direct electrical arc ignition of hybrid rocket motors

    Science.gov (United States)

    Judson, Michael I., Jr.

    Hybrid rockets motors provide distinct safety advantages when compared to traditional liquid or solid propellant systems, due to the inherent stability and relative inertness of the propellants prior to established combustion. As a result of this inherent propellant stability, hybrid motors have historically proven difficult to ignite. State of the art hybrid igniter designs continue to require solid or liquid reactants distinct from the main propellants. These ignition methods however, reintroduce to the hybrid propulsion system the safety and complexity disadvantages associated with traditional liquid or solid propellants. The results of this study demonstrate the feasibility of a novel direct electrostatic arc ignition method for hybrid motors. A series of small prototype stand-alone thrusters demonstrating this technology were successfully designed and tested using Acrylonitrile Butadiene Styrene (ABS) plastic and Gaseous Oxygen (GOX) as propellants. Measurements of input voltage and current demonstrated that arc-ignition will occur using as little as 10 watts peak power and less than 5 joules total energy. The motor developed for the stand-alone small thruster was adapted as a gas generator to ignite a medium-scale hybrid rocket motor using nitrous oxide /and HTPB as propellants. Multiple consecutive ignitions were performed. A large data set as well as a collection of development `lessons learned' were compiled to guide future development and research. Since the completion of this original groundwork research, the concept has been developed into a reliable, operational igniter system for a 75mm hybrid motor using both gaseous oxygen and liquid nitrous oxide as oxidizers. A development map of the direct spark ignition concept is presented showing the flow of key lessons learned between this original work and later follow on development.

  18. High-resolution, label-free imaging of living cells with direct electron-beam-excitation-assisted optical microscopy.

    Science.gov (United States)

    Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu

    2015-06-01

    High spatial resolution microscope is desired for deep understanding of cellular functions, in order to develop medical technologies. We demonstrate high-resolution imaging of un-labelled organelles in living cells, in which live cells on a 50 nm thick silicon nitride membrane are imaged by autofluorescence excited with a focused electron beam through the membrane. Electron beam excitation enables ultrahigh spatial resolution imaging of organelles, such as mitochondria, nuclei, and various granules. Since the autofluorescence spectra represent molecular species, this microscopy allows fast and detailed investigations of cellular status in living cells.

  19. Transcriptome- Assisted Label-Free Quantitative Proteomics Analysis Reveals Novel Insights into Piper nigrum—Phytophthora capsici Phytopathosystem

    Science.gov (United States)

    Mahadevan, Chidambareswaren; Krishnan, Anu; Saraswathy, Gayathri G.; Surendran, Arun; Jaleel, Abdul; Sakuntala, Manjula

    2016-01-01

    Black pepper (Piper nigrum L.), a tropical spice crop of global acclaim, is susceptible to Phytophthora capsici, an oomycete pathogen which causes the highly destructive foot rot disease. A systematic understanding of this phytopathosystem has not been possible owing to lack of genome or proteome information. In this study, we explain an integrated transcriptome-assisted label-free quantitative proteomics pipeline to study the basal immune components of black pepper when challenged with P. capsici. We report a global identification of 532 novel leaf proteins from black pepper, of which 518 proteins were functionally annotated using BLAST2GO tool. A label-free quantitation of the protein datasets revealed 194 proteins common to diseased and control protein datasets of which 22 proteins showed significant up-regulation and 134 showed significant down-regulation. Ninety-three proteins were identified exclusively on P. capsici infected leaf tissues and 245 were expressed only in mock (control) infected samples. In-depth analysis of our data gives novel insights into the regulatory pathways of black pepper which are compromised during the infection. Differential down-regulation was observed in a number of critical pathways like carbon fixation in photosynthetic organism, cyano-amino acid metabolism, fructose, and mannose metabolism, glutathione metabolism, and phenylpropanoid biosynthesis. The proteomics results were validated with real-time qRT-PCR analysis. We were also able to identify the complete coding sequences for all the proteins of which few selected genes were cloned and sequence characterized for further confirmation. Our study is the first report of a quantitative proteomics dataset in black pepper which provides convincing evidence on the effectiveness of a transcriptome-based label-free proteomics approach for elucidating the host response to biotic stress in a non-model spice crop like P. nigrum, for which genome information is unavailable. Our dataset

  20. Transcriptome- Assisted Label-Free Quantitative Proteomics Analysis Reveals Novel Insights into Piper nigrum-Phytophthora capsici Phytopathosystem.

    Science.gov (United States)

    Mahadevan, Chidambareswaren; Krishnan, Anu; Saraswathy, Gayathri G; Surendran, Arun; Jaleel, Abdul; Sakuntala, Manjula

    2016-01-01

    Black pepper (Piper nigrum L.), a tropical spice crop of global acclaim, is susceptible to Phytophthora capsici, an oomycete pathogen which causes the highly destructive foot rot disease. A systematic understanding of this phytopathosystem has not been possible owing to lack of genome or proteome information. In this study, we explain an integrated transcriptome-assisted label-free quantitative proteomics pipeline to study the basal immune components of black pepper when challenged with P. capsici. We report a global identification of 532 novel leaf proteins from black pepper, of which 518 proteins were functionally annotated using BLAST2GO tool. A label-free quantitation of the protein datasets revealed 194 proteins common to diseased and control protein datasets of which 22 proteins showed significant up-regulation and 134 showed significant down-regulation. Ninety-three proteins were identified exclusively on P. capsici infected leaf tissues and 245 were expressed only in mock (control) infected samples. In-depth analysis of our data gives novel insights into the regulatory pathways of black pepper which are compromised during the infection. Differential down-regulation was observed in a number of critical pathways like carbon fixation in photosynthetic organism, cyano-amino acid metabolism, fructose, and mannose metabolism, glutathione metabolism, and phenylpropanoid biosynthesis. The proteomics results were validated with real-time qRT-PCR analysis. We were also able to identify the complete coding sequences for all the proteins of which few selected genes were cloned and sequence characterized for further confirmation. Our study is the first report of a quantitative proteomics dataset in black pepper which provides convincing evidence on the effectiveness of a transcriptome-based label-free proteomics approach for elucidating the host response to biotic stress in a non-model spice crop like P. nigrum, for which genome information is unavailable. Our dataset

  1. Direct Nanoscale Sensing of the Internal Electric Field in Operating Semiconductor Devices Using Single Electron Spins.

    Science.gov (United States)

    Iwasaki, Takayuki; Naruki, Wataru; Tahara, Kosuke; Makino, Toshiharu; Kato, Hiromitsu; Ogura, Masahiko; Takeuchi, Daisuke; Yamasaki, Satoshi; Hatano, Mutsuko

    2017-02-28

    The electric field inside semiconductor devices is a key physical parameter that determines the properties of the devices. However, techniques based on scanning probe microscopy are limited to sensing at the surface only. Here, we demonstrate the direct sensing of the internal electric field in diamond power devices using single nitrogen-vacancy (NV) centers. The NV center embedded inside the device acts as a nanoscale electric field sensor. We fabricated vertical diamond p-i-n diodes containing the single NV centers. By performing optically detected magnetic resonance measurements under reverse-biased conditions with an applied voltage of up to 150 V, we found a large splitting in the magnetic resonance frequencies. This indicated that the NV center senses the transverse electric field in the space-charge region formed in the i-layer. The experimentally obtained electric field values are in good agreement with those calculated by a device simulator. Furthermore, we demonstrate the sensing of the electric field in different directions by utilizing NV centers with different N-V axes. This direct and quantitative sensing method using an electron spin in a wide-band-gap material provides a way to monitor the electric field in operating semiconductor devices.

  2. Physicochemical Properties of Biopolymer Hydrogels Treated by Direct Electric Current

    Directory of Open Access Journals (Sweden)

    Żaneta Król

    2016-07-01

    Full Text Available The objective of this study was to evaluate the changes within the physicochemical properties of gelatine (2%; 4%; 8%, carrageenan (1.5%; 2%; 2.5% and sodium alginate (0.75%; 1%; 1.25% hydrogels with different sodium chloride concentrations that were triggered by applying direct current (DC of 400 mA for a duration of five minutes. There were three types of gels prepared for the purpose of the study: C, control; H, gels on the basis of hydrosols that were treated with DC; and G, gels treated with DC. In the course of the study, the authors carried out the following analyses: Texture Profile Analysis (TPA, Fourier Transform Infrared spectroscopy (FTIR, Scanning Electron Microscopy (SEM and Swelling Ratio (SR. Furthermore, the color and pH of hydrogels were measured. The FTIR spectra showed that the structures of gelatine, carrageenan and sodium alginate do not significantly change upon applying DC. The results of TPA, SR, color and pH measurement indicate that hydrogels’ properties are significantly dependent on the type of polymer, its concentration and the type of the gel. By changing those parameters, the characteristics of such gels can be additionally tuned, which extends their applicability, e.g., in the food industry. Moreover, the analysis revealed that SR of H gel gelatine after 72 h of storage was 1.84-times higher than SR of the control sample, which indicated that this gel may be considered as a possible component for wound dressing materials.

  3. Label-free optical imaging technologies for rapid translation and use during intraoperative surgical and tumor margin assessment

    Science.gov (United States)

    Boppart, Stephen A.; Brown, J. Quincy; Farah, Camile S.; Kho, Esther; Marcu, Laura; Saunders, Christobel M.; Sterenborg, Henricus J. C. M.

    2018-02-01

    The biannual International Conference on Biophotonics was recently held on April 30 to May 1, 2017, in Fremantle, Western Australia. This continuing conference series brought together key opinion leaders in biophotonics to present their latest results and, importantly, to participate in discussions on the future of the field and what opportunities exist when we collectively work together for using biophotonics for biological discovery and medical applications. One session in this conference, entitled "Tumor Margin Identification: Critiquing Technologies," challenged invited speakers and attendees to review and critique representative label-free optical imaging technologies and their application for intraoperative assessment and guidance in surgical oncology. We are pleased to share a summary in this outlook paper, with the intent to motivate more research inquiry and investigations, to challenge these and other optical imaging modalities to evaluate and improve performance, to spur translation and adoption, and ultimately, to improve the care and outcomes of patients.

  4. Label-free optical-resolution photoacoustic microscopy of superficial microvasculature using a compact visible laser diode excitation

    Science.gov (United States)

    Zeng, Lvming; Piao, Zhonglie; Huang, Shenghai; Jia, Wangcun; Chen, Zhongping

    2015-01-01

    We have developed laser-diode-based optical-resolution photoacoustic microscopy (LD-OR-PAM) of superficial microvasculature which has the desirable properties of being compact, low-cost, and label-free. A 300-mW visible pulsed laser diode was operated at a 405 ± 5 nm wavelength with a pulse energy as low as 52 nJ. By using a 3.6 MHz ultrasound transducer, the system was tested on carbon fibers with a lateral resolution of 0.95 µm and an SNR of 38 dB. The subcutaneous microvasculature on a mouse back was imaged without an exogenous contrast agent which demonstrates the potential of the proposed prototype for skin chromophores. Our eventual goal is to offer a practical and affordable multi-wavelength functional LD-OR-PAM instrument suitable for clinical applications. PMID:26698732

  5. In vivo label-free lymphangiography of cutaneous lymphatic vessels in human burn scars using optical coherence tomography.

    Science.gov (United States)

    Gong, Peijun; Es'haghian, Shaghayegh; Harms, Karl-Anton; Murray, Alexandra; Rea, Suzanne; Wood, Fiona M; Sampson, David D; McLaughlin, Robert A

    2016-12-01

    We present an automated, label-free method for lymphangiography of cutaneous lymphatic vessels in humans in vivo using optical coherence tomography (OCT). This method corrects for the variation in OCT signal due to the confocal function and sensitivity fall-off of a spectral-domain OCT system and utilizes a single-scattering model to compensate for A-scan signal attenuation to enable reliable thresholding of lymphatic vessels. A segment-joining algorithm is then incorporated into the method to mitigate partial-volume effects with small vessels. The lymphatic vessel images are augmented with images of the blood vessel network, acquired from the speckle decorrelation with additional weighting to differentiate blood vessels from the observed high decorrelation in lymphatic vessels. We demonstrate the method with longitudinal scans of human burn scar patients undergoing ablative fractional laser treatment, showing the visualization of the cutaneous lymphatic and blood vessel networks.

  6. In vivo, label-free, three-dimensional quantitative imaging of liver surface using multi-photon microscopy

    International Nuclear Information System (INIS)

    Zhuo, Shuangmu; Yan, Jie; Kang, Yuzhan; Xu, Shuoyu; Peng, Qiwen

    2014-01-01

    Various structural features on the liver surface reflect functional changes in the liver. The visualization of these surface features with molecular specificity is of particular relevance to understanding the physiology and diseases of the liver. Using multi-photon microscopy (MPM), we have developed a label-free, three-dimensional quantitative and sensitive method to visualize various structural features of liver surface in living rat. MPM could quantitatively image the microstructural features of liver surface with respect to the sinuosity of collagen fiber, the elastic fiber structure, the ratio between elastin and collagen, collagen content, and the metabolic state of the hepatocytes that are correlative with the pathophysiologically induced changes in the regions of interest. This study highlights the potential of this technique as a useful tool for pathophysiological studies and possible diagnosis of the liver diseases with further development.

  7. Label-free monitoring of DNA polymerase activity based on a thrombin-binding aptamer G-quadruplex.

    Science.gov (United States)

    Wang, Jing; Liu, Haisheng; Ma, Changbei; Wang, Jun; Zhong, Linxiu; Wu, Kefeng

    2017-04-01

    We have developed a label-free assay for the detection of DNA polymerase activity based on a thrombin-binding aptamer (TBA) G-quadruplex. In the presence of DNA polymerase, the 3'-OH termini of the hairpin substrate are immediately elongated to replace the TBA, which can be recognized quickly by the ThT dye and results in an increase of fluorescence. This method is highly sensitive with a detection limit of 0.1 U/mL. It is simple and cost-effective without any requirement of labeling with a fluorophore-quencher pair. Furthermore, the proposed method can also be applied to analyze the inhibition of DNA polymerase, which clearly indicates that the proposed method can be applied for screening of potential DNA polymerase inhibitors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Assessing Collagen and Elastin Pressure-Dependent Microarchitectures in Live, Human Resistance Arteries by Label-Free Fluorescence Microscopy

    DEFF Research Database (Denmark)

    Bloksgaard, Maria; Thorsted, Bjarne; Brewer, Jonathan R.

    2017-01-01

    The pathogenic contribution of resistance artery remodeling is documented in essential hypertension, diabetes and the metabolic syndrome. Investigations and development of microstructurally motivated mathematical models for understanding the mechanical properties of human resistance arteries...... in health and disease have the potential to aid understanding how disease and medical treatments affect the human microcirculation. To develop these mathematical models, it is essential to decipher the relationship between the mechanical and microarchitectural properties of the microvascular wall....... In this work, we describe an ex vivo method for passive mechanical testing and simultaneous label-free three-dimensional imaging of the microarchitecture of elastin and collagen in the arterial wall of isolated human resistance arteries. The imaging protocol can be applied to resistance arteries of any species...

  9. A label-free fluorescent adenosine triphosphate biosensor via overhanging aptamer-triggered enzyme protection and target recycling amplification.

    Science.gov (United States)

    Wang, Zhaoyin; Zhao, Jian; Dai, Zhihui

    2016-06-20

    Herein, a label-free fluorescent adenosine triphosphate (ATP) aptasensor is fabricated with a DNA hairpin and an overhanging aptamer. In the presence of ATP, the overhanging sequences of the aptamer may form preferred substrates of exo III, and thus trigger the enzyme-assisted amplification, which results in the release of G-rich sequences. Free G-rich sequences subsequently generate an enhanced flourescent signal by binding with thioflavin T. However, if ATP is absent, the overhanging sequence can induce steric hindrance and protect the DNA hairpin against the digestion of exo III, significantly reducing the noise of this biosensor. Accordingly, the signal-to-noise ratio of the sensing system is greatly improved, which ensures the desirable analytical performance of the proposed aptasensor both in pure samples and real samples.

  10. High-throughput label-free image cytometry and image-based classification of live Euglena gracilis

    Science.gov (United States)

    Lei, Cheng; Ito, Takuro; Ugawa, Masashi; Nozawa, Taisuke; Iwata, Osamu; Maki, Masanori; Okada, Genki; Kobayashi, Hirofumi; Sun, Xinlei; Tiamsak, Pimsiri; Tsumura, Norimichi; Suzuki, Kengo; Di Carlo, Dino; Ozeki, Yasuyuki; Goda, Keisuke

    2016-01-01

    We demonstrate high-throughput label-free single-cell image cytometry and image-based classification of Euglena gracilis (a microalgal species) under different culture conditions. We perform it with our high-throughput optofluidic image cytometer composed of a time-stretch microscope with 780-nm resolution and 75-Hz line rate, and an inertial-focusing microfluidic device. By analyzing a large number of single-cell images from the image cytometer, we identify differences in morphological and intracellular phenotypes between E. gracilis cell groups and statistically classify them under various culture conditions including nitrogen deficiency for lipid induction. Our method holds promise for real-time evaluation of culture techniques for E. gracilis and possibly other microalgae in a non-invasive manner. PMID:27446699

  11. Functionalized gold nanostars for label-free detection of PKA phosphorylation using surface-enhanced Raman spectroscopy

    Science.gov (United States)

    He, Shuai; Kah, James C. Y.

    2017-04-01

    Protein phosphorylation controls fundamental biological processes. Dysregulation of protein kinase is associated with a series of human diseases including cancer. Protein kinase A (PKA) activity has been reported to serve as a potential prognostic marker for cancer. To this end, we developed a non-radioactive, rapid, cheap and robust scheme based on surface-enhanced Raman spectroscopy (SERS) for label-free detection of PKA phosphorylation using gold nanostars (AuNS) functionalized with BSA-kemptide. While bovine serum albumin (BSA) proteins stabilized the AuNS, kemptide, which is a high affinity substrate peptide specific for PKA, were phosphorylated in vitro to generate Raman signals that were identified by performing principal component analysis (PCA) on the acquired SERS spectra.

  12. Label-free shotgun proteomics and metabolite analysis reveal a significant metabolic shift during citrus fruit development.

    Science.gov (United States)

    Katz, Ehud; Boo, Kyung Hwan; Kim, Ho Youn; Eigenheer, Richard A; Phinney, Brett S; Shulaev, Vladimir; Negre-Zakharov, Florence; Sadka, Avi; Blumwald, Eduardo

    2011-11-01

    Label-free LC-MS/MS-based shot-gun proteomics was used to quantify the differential protein synthesis and metabolite profiling in order to assess metabolic changes during the development of citrus fruits. Our results suggested the occurrence of a metabolic change during citrus fruit maturation, where the organic acid and amino acid accumulation seen during the early stages of development shifted into sugar synthesis during the later stage of citrus fruit development. The expression of invertases remained unchanged, while an invertase inhibitor was up-regulated towards maturation. The increased expression of sucrose-phosphate synthase and sucrose-6-phosphate phosphatase and the rapid sugar accumulation suggest that sucrose is also being synthesized in citrus juice sac cells during the later stage of fruit development.

  13. The 3D OrbiSIMS-label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power.

    Science.gov (United States)

    Passarelli, Melissa K; Pirkl, Alexander; Moellers, Rudolf; Grinfeld, Dmitry; Kollmer, Felix; Havelund, Rasmus; Newman, Carla F; Marshall, Peter S; Arlinghaus, Henrik; Alexander, Morgan R; West, Andy; Horning, Stevan; Niehuis, Ewald; Makarov, Alexander; Dollery, Colin T; Gilmore, Ian S

    2017-12-01

    We report the development of a 3D OrbiSIMS instrument for label-free biomedical imaging. It combines the high spatial resolution of secondary ion mass spectrometry (SIMS; under 200 nm for inorganic species and under 2 μm for biomolecules) with the high mass-resolving power of an Orbitrap (>240,000 at m/z 200). This allows exogenous and endogenous metabolites to be visualized in 3D with subcellular resolution. We imaged the distribution of neurotransmitters-gamma-aminobutyric acid, dopamine and serotonin-with high spectroscopic confidence in the mouse hippocampus. We also putatively annotated and mapped the subcellular localization of 29 sulfoglycosphingolipids and 45 glycerophospholipids, and we confirmed lipid identities with tandem mass spectrometry. We demonstrated single-cell metabolomic profiling using rat alveolar macrophage cells incubated with different concentrations of the drug amiodarone, and we observed that the upregulation of phospholipid species and cholesterol is correlated with the accumulation of amiodarone.

  14. Enhanced vibrational spectroscopy, intracellular refractive indexing for label-free biosensing and bioimaging by multiband plasmonic-antenna array.

    Science.gov (United States)

    Chen, Cheng-Kuang; Chang, Ming-Hsuan; Wu, Hsieh-Ting; Lee, Yao-Chang; Yen, Ta-Jen

    2014-10-15

    In this study, we report a multiband plasmonic-antenna array that bridges optical biosensing and intracellular bioimaging without requiring a labeling process or coupler. First, a compact plasmonic-antenna array is designed exhibiting a bandwidth of several octaves for use in both multi-band plasmonic resonance-enhanced vibrational spectroscopy and refractive index probing. Second, a single-element plasmonic antenna can be used as a multifunctional sensing pixel that enables mapping the distribution of targets in thin films and biological specimens by enhancing the signals of vibrational signatures and sensing the refractive index contrast. Finally, using the fabricated plasmonic-antenna array yielded reliable intracellular observation was demonstrated from the vibrational signatures and intracellular refractive index contrast requiring neither labeling nor a coupler. These unique features enable the plasmonic-antenna array to function in a label-free manner, facilitating bio-sensing and imaging development. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Benchtop chemistry for the rapid prototyping of label-free biosensors: Transmission localized surface plasmon resonance platforms

    Science.gov (United States)

    Liao, Wei-Ssu; Chen, Xin; Yang, Tinglu; Castellana, Edward T.; Chen, Jixin; Cremer, Paul S.

    2012-01-01

    Herein, a simple label-free biosensor fabrication method is demonstrated based on transmission localized surface plasmon resonance (T-LSPR). The platform, which consists of a silver nanoparticle array, can be prepared in just a few minutes using benchtop chemistry. The array was made by a templating technique in conjunction with the photoreduction of Ag ions from solution. This metal surface was functionalized with biotin-linked thiol ligands for binding streptavidin molecules from solution. For an array of 19 nm diameter silver nanoparticles, a redshift in the T-LSPR spectrum of 24 nm was observed upon protein-ligand binding at saturation. The binding constant was found to be 2 × 1012 M–1. Platforms were also fabricated with silver nanoparticles of 34, 55, and 72 nm diameters. The maximum LSPR wavelength shift was nanoparticle size dependent and the maximum sensitivity was obtained with the smaller nanoparticles. PMID:20408728

  16. Chemical surface modifications for the development of silicon-based label-free integrated optical (IO) biosensors: a review.

    Science.gov (United States)

    Bañuls, María-José; Puchades, Rosa; Maquieira, Ángel

    2013-05-13

    Increasing interest has been paid to label-free biosensors in recent years. Among them, refractive index (RI) optical biosensors enable high density and the chip-scale integration of optical components. This makes them more appealing to help develop lab-on-a-chip devices. Today, many RI integrated optical (IO) devices are made using silicon-based materials. A key issue in their development is the biofunctionalization of sensing surfaces because they provide a specific, sensitive response to the analyte of interest. This review critically discusses the biofunctionalization procedures, assay formats and characterization techniques employed in setting up IO biosensors. In addition, it provides the most relevant results obtained from using these devices for real sample biosensing. Finally, an overview of the most promising future developments in the fields of chemical surface modification and capture agent attachment for IO biosensors follows. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Nanomechanical microcantilever operated in vibration modes with use of RNA aptamer as receptor molecules for label-free detection of HCV helicase.

    Science.gov (United States)

    Hwang, Kyo Seon; Lee, Sang-Myung; Eom, Kilho; Lee, Jeong Hoon; Lee, Yoon-Sik; Park, Jung Ho; Yoon, Dae Sung; Kim, Tae Song

    2007-11-30

    We report the nanomechanical microcantilevers operated in vibration modes (oscillation) with use of RNA aptamers as receptor molecules for label-free detection of hepatitis C virus (HCV) helicase. The nanomechanical detection principle is that the ligand-receptor binding on the microcantilever surface induces the dynamic response change of microcantilevers. We implemented the label-free detection of HCV helicase in the low concentration as much as 100 pg/ml from measuring the dynamic response change of microcantilevers. Moreover, from the recent studies showing that the ligand-receptor binding generates the surface stress on the microcantilever, we estimate the surface stress, on the oscillating microcantilevers, induced by ligand-receptor binding, i.e. binding between HCV helicase and RNA aptamer. In this article, it is suggested that the oscillating microcantilevers with use of RNA aptamers as receptor molecules may enable one to implement the sensitive label-free detection of very small amount of small-scale proteins.

  18. A Microneedle Functionalized with Polyethyleneimine and Nanotubes for Highly Sensitive, Label-Free Quantification of DNA.

    Science.gov (United States)

    Saadat-Moghaddam, Darius; Kim, Jong-Hoon

    2017-08-16

    The accurate measure of DNA concentration is necessary for many DNA-based biological applications. However, the current methods are limited in terms of sensitivity, reproducibility, human error, and contamination. Here, we present a microneedle functionalized with polyethyleneimine (PEI) and single-walled carbon nanotubes (SWCNTs) for the highly sensitive quantification of DNA. The microneedle was fabricated using ultraviolet (UV) lithography and anisotropic etching, and then functionalized with PEI and SWCNTs through a dip coating process. The electrical characteristics of the microneedle change with the accumulation of DNA on the surface. Current-voltage measurements in deionized water were conducted to study these changes in the electrical properties of the sensor. The sensitivity test found the signal to be discernable from the noise level down to 100 attomolar (aM), demonstrating higher sensitivity than currently available UV fluorescence and UV absorbance based methods. A microneedle without any surface modification only had a 100 femtomolar (fM) sensitivity. All measurement results were consistent with fluorescence microscopy.

  19. Label-free reflectance hyperspectral imaging for tumor margin assessment: a pilot study on surgical specimens of cancer patients

    Science.gov (United States)

    Fei, Baowei; Lu, Guolan; Wang, Xu; Zhang, Hongzheng; Little, James V.; Patel, Mihir R.; Griffith, Christopher C.; El-Diery, Mark W.; Chen, Amy Y.

    2017-08-01

    A label-free, hyperspectral imaging (HSI) approach has been proposed for tumor margin assessment. HSI data, i.e., hypercube (x,y,λ), consist of a series of high-resolution images of the same field of view that are acquired at different wavelengths. Every pixel on an HSI image has an optical spectrum. In this pilot clinical study, a pipeline of a machine-learning-based quantification method for HSI data was implemented and evaluated in patient specimens. Spectral features from HSI data were used for the classification of cancer and normal tissue. Surgical tissue specimens were collected from 16 human patients who underwent head and neck (H&N) cancer surgery. HSI, autofluorescence images, and fluorescence images with 2-deoxy-2-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]-D-glucose (2-NBDG) and proflavine were acquired from each specimen. Digitized histologic slides were examined by an H&N pathologist. The HSI and classification method were able to distinguish between cancer and normal tissue from the oral cavity with an average accuracy of 90%±8%, sensitivity of 89%±9%, and specificity of 91%±6%. For tissue specimens from the thyroid, the method achieved an average accuracy of 94%±6%, sensitivity of 94%±6%, and specificity of 95%±6%. HSI outperformed autofluorescence imaging or fluorescence imaging with vital dye (2-NBDG or proflavine). This study demonstrated the feasibility of label-free, HSI for tumor margin assessment in surgical tissue specimens of H&N cancer patients. Further development of the HSI technology is warranted for its application in image-guided surgery.

  20. Ultrasensitive electrochemical aptasensor based on sandwich architecture for selective label-free detection of colorectal cancer (CT26) cells.

    Science.gov (United States)

    Hashkavayi, Ayemeh Bagheri; Raoof, Jahan Bakhsh; Ojani, Reza; Kavoosian, Saeid

    2017-06-15

    Colorectal cancer is one of the most common cancers in the world and has no effective treatment. Therefore, development of new methods for early diagnosis is instantly required. Biological recognition probes such as synthetic receptor and aptamer is one of the candidate recognition layers to detect important biomolecules. In this work, an electrochemical aptasensor was developed by fabricating an aptamer-cell-aptamer sandwich architecture on an SBA-15-3-aminopropyltriethoxysilane (SBA-15-pr-NH 2 ) and Au nanoparticles (AuNPs) modified graphite screen printed electrode (GSPE) surface for the selective, label-free detection of CT26 cancer cells. Based on the incubation of the thiolated aptamer with CT26 cells, the electron-transfer resistance of Fe (CN) 6 3-/4- redox couple increased considerably on the aptasensor surface. The results obtained from cyclic voltammetry and electrochemical impedance spectroscopy studies showed that the fabricated aptasensor can specifically identify CT26 cells in the concentration ranges of 10-1.0×10 5 cells/mL and 1.0×10 5 -6.0×10 6 cells/mL, respectively, with a detection limit of 2cells/mL. Applying the thiol terminated aptamer (5TR1) as a recognition layer led to a sensor with high affinity for CT26 cancer cells, compared to control cancer cells of AGS cells, VERO Cells, PC3 cells and SKOV-3 cells. Therefore a simple, rapid, label free, inexpensive, excellent, sensitive and selective electrochemical aptasensor based on sandwich architecture was developed for detection of CT26 Cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Rapid label-free profiling of oral cancer biomarker proteins using nano-UPLC-Q-TOF ion mobility mass spectrometry.

    Science.gov (United States)

    Nassar, Ala F; Williams, Brad J; Yaworksy, Dustin C; Patel, Vyomesh; Rusling, James F

    2016-03-01

    It has become quite clear that single cancer biomarkers cannot in general provide high sensitivity and specificity for reliable clinical cancer diagnostics. This paper explores the feasibility of rapid detection of multiple biomarker proteins in model oral cancer samples using label-free protein relative quantitation. MS-based label-free quantitative proteomics offer a rapid alternative that bypasses the need for stable isotope containing compounds to chemically bind and label proteins. Total protein content in oral cancer cell culture conditioned media was precipitated, subjected to proteolytic digestion, and then analyzed using a nano-UPLC (where UPLC is ultra-performance liquid chromatography) coupled to a hybrid Q-Tof ion-mobility mass spectrometry (MS). Rapid, simultaneous identification and quantification of multiple possible cancer biomarker proteins was achieved. In a comparative study between cancer and noncancer samples, approximately 952 proteins were identified using a high-throughput 1D ion mobility assisted data independent acquisition (IM-DIA) approach. As we previously demonstrated that interleukin-8 (IL-8) and vascular endothelial growth factor A (VEGF-A) were readily detected in oral cancer cell conditioned media(1), we targeted these biomarker proteins to validate our approach. Target biomarker protein IL-8 was found between 3.5 and 8.8 fmol, while VEGF-A was found at 1.45 fmol in the cancer cell media. Overall, our data suggest that the nano-UPLC-IM-DIA bioassay is a feasible approach to identify and quantify proteins in complex samples without the need for stable isotope labeling. These results have significant implications for rapid tumor diagnostics and prognostics by monitoring proteins such as IL-8 and VEGF-A implicated in cancer development and progression. The analysis in tissue or plasma is not possible at this time, but the subsequent work would be needed for validation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Non-invasive in vivo characterization of skin wound healing using label-free multiphoton microscopy (Conference Presentation)

    Science.gov (United States)

    Jones, Jake D.; Majid, Fariah; Ramser, Hallie; Quinn, Kyle P.

    2017-02-01

    Non-healing ulcerative wounds, such as diabetic foot ulcers, are challenging to diagnose and treat due to their numerous possible etiologies and the variable efficacy of advanced wound care products. Thus, there is a critical need to develop new quantitative biomarkers and diagnostic technologies that are sensitive to wound status in order to guide care. The objective of this study was to evaluate the utility of label-free multiphoton microscopy for characterizing wound healing dynamics in vivo and identifying potential differences in diabetic wounds. We isolated and measured an optical redox ratio of FAD/(NADH+FAD) autofluorescence to provide three-dimensional maps of local cellular metabolism. Using a mouse model of wound healing, in vivo imaging at the wound edge identified a significant decrease in the optical redox ratio of the epidermis (p≤0.0103) between Days 3 through 14 compared to Day 1. This decrease in redox ratio coincided with a decrease in NADH fluorescence lifetime and thickening of the epithelium, collectively suggesting a sensitivity to keratinocyte hyperproliferation. In contrast to normal wounds, we have found that keratinocytes from diabetic wounds remain in a proliferative state at later time points with a lower redox ratio at the wound edge. Microstructural organization and composition was also measured from second harmonic generation imaging of collagen and revealed differences between diabetic and non-diabetic wounds. Our work demonstrates label-free multiphoton microscopy offers potential to provide non-invasive structural and functional biomarkers associated with different stages of skin wound healing, which may be used to detect delayed healing and guide treatment.

  3. Label-free separation of human embryonic stem cells (hESCs) and their cardiac derivatives using Raman spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Chan, J W; Lieu, D K; Huser, T R; Li, R A

    2008-09-08

    Self-renewable, pluripotent human embryonic stem cells (hESCs) can be differentiated into cardiomyocytes (CMs), providing an unlimited source of cells for transplantation therapies. However, unlike certain cell lineages such as hematopoietic cells, CMs lack specific surface markers for convenient identification, physical separation, and enrichment. Identification by immunostaining of cardiac-specific proteins such as troponin requires permeabilization, which renders the cells unviable and non-recoverable. Ectopic expression of a reporter protein under the transcriptional control of a heart-specific promoter for identifying hESC-derived CMs (hESC-CMs) is useful for research but complicates potential clinical applications. The practical detection and removal of undifferentiated hESCs in a graft, which may lead to tumors, is also critical. Here, we demonstrate a non-destructive, label-free optical method based on Raman scattering to interrogate the intrinsic biochemical signatures of individual hESCs and their cardiac derivatives, allowing cells to be identified and classified. By combining the Raman spectroscopic data with multivariate statistical analysis, our results indicate that hESCs, human fetal left ventricular CMs, and hESC-CMs can be identified by their intrinsic biochemical characteristics with an accuracy of 96%, 98% and 66%, respectively. The present study lays the groundwork for developing a systematic and automated method for the non-invasive and label-free sorting of (i) high-quality hESCs for expansion, and (ii) ex vivo CMs (derived from embryonic or adult stem cells) for cell-based heart therapies.

  4. An ultrasensitive label-free electrochemiluminescent immunosensor for measuring Cry1Ab level and genetically modified crops content.

    Science.gov (United States)

    Gao, Hongfei; Wen, Luke; Wu, Yuhua; Fu, Zhifeng; Wu, Gang

    2017-11-15

    The development of genetically modified (GM) insect-resistant crops has aroused great public concern about the risks on the eco-environment resulting from a release of toxic Cry proteins (such as Cry1Ab) to the soil. Therefore, it is of crucial importance to measure the Cry proteins level and the GM crops content. Here, we have tested for the first time a method that uses novel carbon nanospheres (CNPs) label-free electrochemiluminescent (ECL) immunosensor for the ultrasensitive quantification of Cry1Ab and GM crops. In this work, novel CNPs were prepared from printer toner with a very facile approach, and linked with anti-Cry1Ab antibodies to modify a golden working electrode. The immunoreaction between Cry1Ab and its antibody formed an immunocomplex on the bioreceptor region of the sensor, which inhibited electron transfer between the electrode surface and the ECL substance, leading to a decrease of ECL response. Under the optimal conditions, the fabricated label-free ECL immunosensor determined Cry1Ab down to 3.0pgmL -1 within a linear range of 0.010-1.0ngmL -1 , showing significant improvement of sensitivity than that of most previous reports. Meanwhile, the proposed method was successfully applied for GM rice BT63 and GM maize MON810 detections down to 0.010% and 0.020%, respectively. Due to its outstanding advantages such as high sensitivity, ideal selectivity, simple fabrication, rapid detection, and low cost, the developed method can be considered as a powerful and pioneering tool for GM crops detection. Its use can also be extended to other toxin protein sensing in foods. Copyright © 2017. Published by Elsevier B.V.

  5. Optical integrated chips with micro and nanostructures for refractive index and SERS-based optical label-free sensing

    Directory of Open Access Journals (Sweden)

    Liu Liu

    2015-11-01

    Full Text Available Label-free optical biosensing technologies have superior abilities of quantitative analysis, unmodified targets, and ultrasmall sample volume, compared to conventional fluorescence-label-based sensing techniques, in detecting various biomolecules. In this review article, we introduce our recent results in the field of evanescent-wavebased refractive index sensing and surface enhanced Raman scattering (SERS-based sensing, both of which are promising platforms for label-free optical biosensors. First, silicon-on-insulator (SOI nanowire waveguide and metallic surface plasmon resonance (SPR-based refractive index sensing are discussed. In order to improve the detection limit, phase interrogation techniques are introduced to these types of sensors based on prism-coupled SPR and SOI microring resonators. A detection limit in the order of 10−6 refractive index unit is achieved. Detection of 16.7 pM anti-IgG is also demonstrated based on the SPR devices. Second, SERS substrates based on various nanometallic structures are discussed. Metallic nanowire arrays and inverted nanopyramids and grooves with a thin metal surface are fabricated based on anisotropic wetetching of silicon substrates. Both structures have demonstrated a Raman signal enhancement on the order of 107. In order to improve the extraction efficiency of the Raman signal at a high wave number, a nano-bowtie array substrate is fabricated, which exhibits double resonances at both the excitation wavelength and the desired Raman scattering wavelength. Experimental results have shown that this double-resonance structure can further enhance the received Raman signal, as compared to conventional SERS substrates with only one resonance at the excitation wavelength.

  6. Label-Free Proteomic Identification of Endogenous, Insulin-Stimulated Interaction Partners of Insulin Receptor Substrate-1

    Science.gov (United States)

    Geetha, Thangiah; Langlais, Paul; Luo, Moulun; Mapes, Rebekka; Lefort, Natalie; Chen, Shu-Chuan; Mandarino, Lawrence J.; Yi, Zhengping

    2011-03-01

    Protein-protein interactions are key to most cellular processes. Tandem mass spectrometry (MS/MS)-based proteomics combined with co-immunoprecipitation (CO-IP) has emerged as a powerful approach for studying protein complexes. However, a majority of systematic proteomics studies on protein-protein interactions involve the use of protein overexpression and/or epitope-tagged bait proteins, which might affect binding stoichiometry and lead to higher false positives. Here, we report an application of a straightforward, label-free CO-IP-MS/MS method, without the use of protein overexpression or protein tags, to the investigation of changes in the abundance of endogenous proteins associated with a bait protein, which is in this case insulin receptor substrate-1 (IRS-1), under basal and insulin stimulated conditions. IRS-1 plays a central role in the insulin signaling cascade. Defects in the protein-protein interactions involving IRS-1 may lead to the development of insulin resistance and type 2 diabetes. HPLC-ESI-MS/MS analyses identified eleven novel endogenous insulin-stimulated IRS-1 interaction partners in L6 myotubes reproducibly, including proteins play an important role in protein dephosphorylation [protein phosphatase 1 regulatory subunit 12A, (PPP1R12A)], muscle contraction and actin cytoskeleton rearrangement, endoplasmic reticulum stress, and protein folding, as well as protein synthesis. This novel application of label-free CO-IP-MS/MS quantification to assess endogenous interaction partners of a specific protein will prove useful for understanding how various cell stimuli regulate insulin signal transduction.

  7. A Rapid Label-Free Fluorescent Aptasensor PicoGreen-Based Strategy for Aflatoxin B₁ Detection in Traditional Chinese Medicines.

    Science.gov (United States)

    Zhang, Cheng; Dou, Xiaowen; Zhang, Lei; Sun, Meifeng; Zhao, Ming; OuYang, Zhen; Kong, Dandan; Antonio, F Logrieco; Yang, Meihua

    2018-02-28

    Aflatoxin B₁ (AFB₁) is a very hazardous carcinogen, readily contaminating foodstuffs and traditional Chinese medicines (TCMs) that has inspired increasing health concerns due to dietary exposure. Colloidal nanocrystals have been proposed as optical labels for aptasensor assembly, but these typically require tedious multistep conjugation and suffer from unsatisfactory robustness when used for complex matrices. In the present study, we report a rapid and sensitive method for screening for trace AFB₁ levels in TCMs using a label-free fluorescent aptasensor PicoGreen dye-based strategy. Using PicoGreen to selectively measure complementary double-stranded DNA, fluorescence enhancement due to dsDNA is 'turned off' in the presence of AFB₁ due binding of aptamer target over complementary sequence. Self-assembly of a label-free fluorescent aptasensor based on AFB₁ aptamer and PicoGreen dye was performed. Due to competition between the complementary sequence and AFB₁ target, this rapid method was capable of highly sensitive and selective screening for AFB₁ in five types of TCMs. This proposed approach had a limit of detection as low as 0.1 μg·L -1 and good linearity with a range of 0.1-10 μg·L -1 (0.1-10 ppb). Among the 20 samples tested, 6 batches were found to be contaminated with AFB₁ using this method, which was confirmed using sophisticated liquid chromatography-electrospray ionization-tandem mass spectrometry/mass spectrometry analysis. The results of this study indicate the developed method has the potential to be a simple, quick, and sensitive tool for detecting AFB₁ in TCMs.

  8. Optical integrated chips with micro and nanostructures for refractive index and SERS-based optical label-free sensing

    Science.gov (United States)

    Liu, Liu; Jin, Mingliang; Shi, Yaocheng; Lin, Jiao; Zhang, Yuan; Jiang, Li; Zhou, Guofu; He, Sailing

    2015-11-01

    Label-free optical biosensing technologies have superior abilities of quantitative analysis, unmodified targets, and ultrasmall sample volume, compared to conventional fluorescence-label-based sensing techniques, in detecting various biomolecules. In this review article, we introduce our recent results in the field of evanescent-wavebased refractive index sensing and surface enhanced Raman scattering (SERS)-based sensing, both of which are promising platforms for label-free optical biosensors. First, silicon-on-insulator (SOI) nanowire waveguide and metallic surface plasmon resonance (SPR)-based refractive index sensing are discussed. In order to improve the detection limit, phase interrogation techniques are introduced to these types of sensors based on prism-coupled SPR and SOI microring resonators. A detection limit in the order of 10-6 refractive index unit is achieved. Detection of 16.7 pM anti-IgG is also demonstrated based on the SPR devices. Second, SERS substrates based on various nanometallic structures are discussed. Metallic nanowire arrays and inverted nanopyramids and grooves with a thin metal surface are fabricated based on anisotropic wetetching of silicon substrates. Both structures have demonstrated a Raman signal enhancement on the order of 107. In order to improve the extraction efficiency of the Raman signal at a high wave number, a nano-bowtie array substrate is fabricated, which exhibits double resonances at both the excitation wavelength and the desired Raman scattering wavelength. Experimental results have shown that this double-resonance structure can further enhance the received Raman signal, as compared to conventional SERS substrates with only one resonance at the excitation wavelength.

  9. Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements.

    Science.gov (United States)

    Khosravi, Farhad; Loeian, Seyed Masoud; Panchapakesan, Balaji

    2017-04-17

    This study demonstrates the rapid and label-free detection of Interleukin-6 (IL-6) using carbon nanotube micro-arrays with aptamer as the molecular recognition element. Single wall carbon nanotubes micro-arrays biosensors were manufactured using photo-lithography, metal deposition, and etching techniques. Nanotube biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers. Real time response of the sensor conductance was monitored with increasing concentration of IL-6 (1 pg/mL to 10 ng/mL), exposure to the sensing surface in buffer solution, and clinically relevant spiked blood samples. Non-specific Bovine Serum Albumin (BSA), PBS samples, and anti-IgG functionalized devices gave similar signatures in the real time conductance versus time experiments with no significant change in sensor signal. Exposure of the aptamer functionalized nanotube surface to IL-6 decreased the conductance with increasing concentration of IL-6. Experiments based on field effect transistor arrays suggested shift in drain current versus gate voltage for 1 pg and 1 ng of IL-6 exposure. Non-specific BSA did not produce any appreciable shift in the I ds versus V g suggesting specific interactions of IL-6 on PASE conjugated aptamer surface gave rise to the change in electrical signal. Both Z axis and phase image in an Atomic Force Microscope (AFM) suggested unambiguous molecular interaction of the IL-6 on the nanotube-aptamer surface at 1 pg/mL concentration. The concentration of 1 pg falls below the diagnostic gray zone for cancer (2.3 pg-4 ng/mL), which is an indicator of early stage cancer. Thus, nanotube micro-arrays could potentially be developed for creating multiplexed assays involving cancer biomarker proteins and possibly circulating tumor cells all in a single assay using PASE functionalization protocol.

  10. Label-free cellular structure imaging with 82 nm lateral resolution using an electron-beam excitation-assisted optical microscope.

    Science.gov (United States)

    Fukuta, Masahiro; Masuda, Yuriko; Inami, Wataru; Kawata, Yoshimasa

    2016-07-25

    We present label-free and high spatial-resolution imaging for specific cellular structures using an electron-beam excitation-assisted optical microscope (EXA microscope). Images of the actin filament and mitochondria of stained HeLa cells, obtained by fluorescence and EXA microscopy, were compared to identify cellular structures. Based on these results, we demonstrated the feasibility of identifying label-free cellular structures at a spatial resolution of 82 nm. Using numerical analysis, we calculated the imaging depth region and determined the spot size of a cathodoluminescent (CL) light source to be 83 nm at the membrane surface.

  11. Label-free DNA electrochemical sensor based on a PNA-functionalized conductive polymer

    DEFF Research Database (Denmark)

    Reisberg, S; Dang, L A; Nguyen, Q A

    2008-01-01

    -solution interface. A reagentless and direct electrochemical detection was obtained by detection of the electrochemical changes using square wave voltammetry (SWV). An increase in the peak current of quinone was observed upon hybridization of probe on the target, whereas no change is observed with non...

  12. Mutagenic and genotoxic potential of direct electric current in Escherichia coli and Salmonella thyphimurium strains.

    Science.gov (United States)

    Gomes, Marina das Neves; Cardoso, Janine Simas; Leitão, Alvaro Costa; Quaresma, Carla Holandino

    2016-05-01

    Direct electric current has several therapeutic uses such as antibacterial and antiprotozoal action, tissues scarring and regeneration, as well as tumor treatment. This method has shown promising results in vivo and in vitro, with significant efficacy and almost no side effects. Considering lack of studies regarding direct electric current mutagenic and/or genotoxic effects, the present work evaluated both aspects by using five different bacterial experimental assays: survival of repair-deficient mutants, Salmonella-histidine reversion mutagenesis (Ames test), forward mutations to rifampicin resistance, phage reactivation, and lysogenic induction. In these experimental conditions, cells were submitted to an approach that allows evaluation of anodic, cathodic, and electro-ionic effects generated by 2 mA of direct electric current, with doses ranging from 0.36 to 3.60 Coulombs. Our results showed these doses did not induce mutagenic or genotoxic effects. © 2016 Wiley Periodicals, Inc.

  13. The difference between electrical microstimulation and direct electrical stimulation - towards new opportunities for innovative functional brain mapping?

    Science.gov (United States)

    Vincent, Marion; Rossel, Olivier; Hayashibe, Mitsuhiro; Herbet, Guillaume; Duffau, Hugues; Guiraud, David; Bonnetblanc, François

    2016-04-01

    Both electrical microstimulation (EMS) and direct electrical stimulation (DES) of the brain are used to perform functional brain mapping. EMS is applied to animal fundamental neuroscience experiments, whereas DES is performed in the operating theatre on neurosurgery patients. The objective of the present review was to shed new light on electrical stimulation techniques in brain mapping by comparing EMS and DES. There is much controversy as to whether the use of DES during wide-awake surgery is the 'gold standard' for studying the brain function. As part of this debate, it is sometimes wrongly assumed that EMS and DES induce similar effects in the nervous tissues and have comparable behavioural consequences. In fact, the respective stimulation parameters in EMS and DES are clearly different. More surprisingly, there is no solid biophysical rationale for setting the stimulation parameters in EMS and DES; this may be due to historical, methodological and technical constraints that have limited the experimental protocols and prompted the use of empirical methods. In contrast, the gap between EMS and DES highlights the potential for new experimental paradigms in electrical stimulation for functional brain mapping. In view of this gap and recent technical developments in stimulator design, it may now be time to move towards alternative, innovative protocols based on the functional stimulation of peripheral nerves (for which a more solid theoretical grounding exists).

  14. A theoretical study of the possible use of electroosmotic flow to extend the read length of DNA sequencing by end-labeled free solution electrophoresis.

    Science.gov (United States)

    McCormick, Laurette C; Slater, Gary W

    2006-05-01

    End-labeled free solution electrophoresis (ELFSE) provides a means of separating DNA with free-solution CE, eliminating the need for gels and polymer solutions which increase the run time and can be difficult to load into a capillary. In free-solution electrophoresis, DNA is normally free-draining and all fragments reach the detector at the same time, whereas ELFSE uses an uncharged label molecule attached to each DNA fragment in order to render the electrophoretic mobility size-dependent. With ELFSE, however, the larger molecules are not separated enough (limiting the read length in the case of ssDNA sequencing) while the smaller ones are overseparated; the larger ones are too fast while the shorter ones are too slow, which is the opposite of traditional gel-based methods. In this article, we show how an EOF could be used to overcome these problems and extend the DNA sequencing read length of ELFSE. This counterflow would allow the larger, previously unresolved molecules more time to separate and thereby increase the read length. Through our theoretical investigation, we predict that an EOF mobility of approximately the same magnitude as that of unlabeled DNA would provide the best results for the regime where all molecules move in the same direction. Even better resolution would be possible for smaller values of EOF which allow different directions of migration; however, the migration times then would become too large. The flow would need to be well controlled since the gain in read length decreases as the magnitude of the counterflow increases; an EOF mobility double that of unlabeled DNA would no longer increase the read length, although ELFSE would still benefit from a reduction in migration time.

  15. Analysis of critical thinking ability in direct current electrical problems solving

    Science.gov (United States)

    Hartono; Sunarno, Widha; Sarwanto; Arya Nugraha, Dewanta

    2017-11-01

    This study concern on analyzing the ability of students in critical thinking skills on the subject matter of direct current electricity. Samples were taken using purposive random sampling consisted of 32 students of grade XI, Multimedia 1, SMK Negeri 3 Surakarta in academic year 2016/2017. This study used descriptive quantitative method. The data were collected using tests and interviews regarding the subject matter of direct current electricity. Based on the results, students are getting some difficulties in solving problem in indicator 4. The average of students’ correct answer is 62.8%.

  16. Research Capabilities Directed to all Electric Engineering Teachers, from an Alternative Energy Model

    Directory of Open Access Journals (Sweden)

    Víctor Hugo Ordóñez Navea

    2017-08-01

    Full Text Available The purpose of this work was to contemplate research capabilities directed to all electric engineering teachers from an alternative energy model intro the explanation of a semiconductor in the National Training Program in Electricity. Some authors, such as. Vidal (2016, Atencio (2014 y Camilo (2012 point out to technological applications with semiconductor electrical devices. In this way; a diagnostic phase is presented, held on this field research as a descriptive type about: a how to identify the necessities of alternative energies, and b The research competences in the alternatives energies of researcher from a solar cell model, to boost and innovate the academic praxis and technologic ingenuity. Themselves was applied a survey for a group of 15 teachers in the National Program of Formation in electricity to diagnose the deficiencies in the research area of alternatives energies. The process of data analysis was carried out through descriptive statistic. Later the conclusions are presented the need to generate strategies for stimulate and propose exploration of alternatives energies to the development of research competences directed to the teachers of electrical engineering for develop the research competences in the enforcement of the teachers exercise for the electric engineering, from an alternative energy model and boost the technologic research in the renewal energies field.

  17. Blood Stage Plasmodium falciparum Exhibits Biological Responses to Direct Current Electric Fields.

    Directory of Open Access Journals (Sweden)

    Lorena M Coronado

    Full Text Available The development of resistance to insecticides by the vector of malaria and the increasingly faster appearance of resistance to antimalarial drugs by the parasite can dangerously hamper efforts to control and eradicate the disease. Alternative ways to treat this disease are urgently needed. Here we evaluate the in vitro effect of direct current (DC capacitive coupling electrical stimulation on the biology and viability of Plasmodium falciparum. We designed a system that exposes infected erythrocytes to different capacitively coupled electric fields in order to evaluate their effect on P. falciparum. The effect on growth of the parasite, replication of DNA, mitochondrial membrane potential and level of reactive oxygen species after exposure to electric fields demonstrate that the parasite is biologically able to respond to stimuli from DC electric fields involving calcium signaling pathways.

  18. Bi-directional causality in California's electricity and natural-gas markets

    International Nuclear Information System (INIS)

    Woo, Chi-Keung; Olson, Arne; Horowitz, Ira; Luk, Stephen

    2006-01-01

    The Granger instantaneous-causality test is applied to explore the potential causal relationships between wholesale electricity and natural-gas prices in California. The test shows these relationships to be bi-directional, and reveals California's electricity and natural-gas markets to be as inextricably intertwined as casual observation and theoretical considerations would suggest they ought to be. This meshing of markets exacerbated the effects of California's natural-gas crisis on the contemporaneous electricity crisis, while concurrently the electricity crisis may have contributed to the dysfunction in the national-gas market and helped to precipitate the natural-gas crisis. The finding supports an integrated approach, as opposed to a piecemeal approach, for formulating energy policy recommendations, not just in California but in the world at large

  19. Disulfide-induced self-assembled targets : A novel strategy for the label free colorimetric detection of DNAs/RNAs via unmodified gold nanoparticles

    NARCIS (Netherlands)

    Shokri, Ehsan; Hosseini, Morteza; Davari, Mehdi D.; Ganjali, Mohammad R.; Peppelenbosch, Maikel P.; Rezaee, Farhad

    2017-01-01

    A modified non-cross-linking gold-nanoparticles (Au-NPs) aggregation strategy has been developed for the label free colorimetric detection of DNAs/RNAs based on self-assembling target species in the presence of thiolated probes. Two complementary thiol-modified probes, each of which specifically

  20. Disulfide-induced self-assembled targets: A novel strategy for the label free colorimetric detection of DNAs/RNAs via unmodified gold nanoparticles

    NARCIS (Netherlands)

    Shokri, E. (Ehsan); M. Hosseini (Morteza); Davari, M.D. (Mehdi D.); Ganjali, M.R. (Mohammad R.); M.P. Peppelenbosch (Maikel); F. Rezaee (Farhad)

    2017-01-01

    textabstractA modified non-cross-linking gold-nanoparticles (Au-NPs) aggregation strategy has been developed for the label free colorimetric detection of DNAs/RNAs based on self-assembling target species in the presence of thiolated probes. Two complementary thiol- modified probes, each of which

  1. Differentiating intratumoral melanocytes from Langerhans cells in nonmelanocytic pigmented skin tumors in vivo by label-free third-harmonic generation microscopy

    Science.gov (United States)

    Weng, Wei-Hung; Liao, Yi-Hua; Tsai, Ming-Rung; Wei, Ming-Liang; Huang, Hsin-Yi; Sun, Chi-Kuang

    2016-07-01

    Morphology and distribution of melanocytes are critical imaging information for the diagnosis of melanocytic lesions. However, how to image intratumoral melanocytes noninvasively in pigmented skin tumors is seldom investigated. Third-harmonic generation (THG) is shown to be enhanced by melanin, whereas high accuracy has been demonstrated using THG microscopy for in vivo differential diagnosis of nonmelanocytic pigmented skin tumors. It is thus desirable to investigate if label-free THG microscopy was capable to in vivo identify intratumoral melanocytes. In this study, histopathological correlations of label-free THG images with the immunohistochemical images stained with human melanoma black (HMB)-45 and cluster of differentiation 1a (CD1a) were made. The correlation results indicated that the intratumoral THG-bright dendritic-cell-like signals were endogenously derived from melanocytes rather than Langerhans cells (LCs). The consistency between THG-bright dendritic-cell-like signals and HMB-45 melanocyte staining showed a kappa coefficient of 0.807, 84.6% sensitivity, and 95% specificity. In contrast, a kappa coefficient of -0.37, 21.7% sensitivity, and 30% specificity were noted between the THG-bright dendritic-cell-like signals and CD1a staining for LCs. Our study indicates the capability of noninvasive label-free THG microscopy to differentiate intratumoral melanocytes from LCs, which is not feasible in previous in vivo label-free clinical-imaging modalities.

  2. Molecular recognition of carboxylates in the protein leucine zipper by a multivalent supramolecular ligand: residue-specific, sensitive and label-free probing by UV resonance Raman spectroscopy.

    Science.gov (United States)

    Zakeri, B; Niebling, S; Martinéz, A G; Sokkar, P; Sanchez-Garcia, E; Schmuck, C; Schlücker, S

    2018-01-17

    Ultraviolet resonance Raman (UVRR) spectroscopy is a selective, sensitive and label-free vibrational spectroscopic technique. Here, we demonstrate as proof of concept that UVRR can be used for probing the recognition between a multivalent supramolecular ligand and acidic residues in leucine zipper, an α-helical structural motif of many proteins.

  3. Electric Vehicles in Colorado: Anticipating Consumer Demand for Direct Current Fast Charging

    Energy Technology Data Exchange (ETDEWEB)

    Wood, Eric W. [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Rames, Clement L. [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-07-01

    To support the State of Colorado in planning for growth in direct current fast charging (DCFC) for electric vehicles, the National Renewable Energy Laboratory (NREL) has partnered with the Regional Air Quality Council (RAQC) and the Colorado Department of Transportation (CDOT) to analyze a number of DCFC investment scenarios. NREL analyzed existing electric vehicle registration data from IHS Markit (IHS) to highlight early trends in the electric vehicle market, which were compared with sales forecasts predicting large growth in the Colorado electric vehicle market. Electric vehicle forecasts were then used to develop future DCFC scenarios to be evaluated in a simulation environment to estimate consumer benefits of the hypothetical DCFC networks in terms of increased driving range and electric vehicle miles traveled (eVMT). Simulated utilization of the hypothetical DCFC networks was analyzed for geographic trends, particularly for correlations with vehicle electric range. Finally, a subset of simulations is presented for consumers with potentially inconsistent access to charging at their home location and presumably greater reliance on public DCFC infrastructure.

  4. Electric Power Generation through the Direct Interaction of Pristine Graphene-Oxide with Water Molecules.

    Science.gov (United States)

    Xu, Tong; Ding, Xiaoteng; Shao, Changxiang; Song, Long; Lin, Tengyu; Gao, Xue; Xue, Jiangli; Zhang, Zhipan; Qu, Liangti

    2018-02-26

    Converting ubiquitous environmental energy into electric power holds tremendous social and financial interests. Traditional energy harvesters and converters are limited by the specific materials and complex configuration of devices. Herein, it is presented that electric power can be directly produced from pristine graphene oxide (GO) without any pretreatment or additives once encountering the water vapor, which will generate an open-circuit-voltage of up to 0.4-0.7 V and a short-circuit-current-density of 2-25 µA cm -2 on a single piece of GO film. This phenomenon results from the directional movement of charged hydrogen ions through the GO film. The present work demonstrates and provides an extremely simple method for electric energy generation, which offers more applications of graphene-based materials in green energy converting field. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Label-free SERS detection of relevant bioanalytes on silver-coated carbon nanotubes: The case of cocaine

    Science.gov (United States)

    Sanles-Sobrido, Marcos; Rodríguez-Lorenzo, Laura; Lorenzo-Abalde, Silvia; González-Fernández, África; Correa-Duarte, Miguel A.; Alvarez-Puebla, Ramón A.; Liz-Marzán, Luis M.

    2009-09-01

    Surface-enhanced Raman scattering (SERS) spectroscopy can be used for the label-free determination and quantification of relevant small biometabolites that are hard to identify by conventional immunological methods, in the absence of labelling. In this work, detection is based on monitoring the vibrational changes occurring at a specific biointerface (a monoclonal antibody, mAb) supported on silver-coated carbon nanotubes (CNT@Ag). Engineered CNT@Ag play a key role, as they offer a stable substrate to support the biointerface, with a high density of hot spots. Proof of concept is demonstrated through the analysis and quantification of the main cocaine metabolite benzoylecgonine. These results open a new avenue toward the generation of portable sensors for fast ultradetection and quantification of relevant metabolites. The use of discrete particles (CNT@Ag@mAb) rather than rough films, or other conventional SERS supports, will also enable a safe remote interrogation of highly toxic sources in environmental problems or in biological fluids.Surface-enhanced Raman scattering (SERS) spectroscopy can be used for the label-free determination and quantification of relevant small biometabolites that are hard to identify by conventional immunological methods, in the absence of labelling. In this work, detection is based on monitoring the vibrational changes occurring at a specific biointerface (a monoclonal antibody, mAb) supported on silver-coated carbon nanotubes (CNT@Ag). Engineered CNT@Ag play a key role, as they offer a stable substrate to support the biointerface, with a high density of hot spots. Proof of concept is demonstrated through the analysis and quantification of the main cocaine metabolite benzoylecgonine. These results open a new avenue toward the generation of portable sensors for fast ultradetection and quantification of relevant metabolites. The use of discrete particles (CNT@Ag@mAb) rather than rough films, or other conventional SERS supports, will also

  6. Uptake of label-free graphene oxide by Caco-2 cells is dependent on the cell differentiation status.

    Science.gov (United States)

    Kucki, Melanie; Diener, Liliane; Bohmer, Nils; Hirsch, Cordula; Krug, Harald F; Palermo, Vincenzo; Wick, Peter

    2017-06-21

    Understanding the interaction of graphene-related materials (GRM) with human cells is a key to the assessment of their potential risks for human health. There is a knowledge gap regarding the potential uptake of GRM by human intestinal cells after unintended ingestion. Therefore the aim of our study was to investigate the interaction of label-free graphene oxide (GO) with the intestinal cell line Caco-2 in vitro and to shed light on the influence of the cell phenotype given by the differentiation status on cellular uptake behaviour. Internalisation of two label-free GOs with different lateral size and thickness by undifferentiated and differentiated Caco-2 cells was analysed by scanning electron microscopy and transmission electron microscopy. Semi-quantification of cells associated with GRM was performed by flow cytometry. Undifferentiated Caco-2 cells showed significant amounts of cell-associated GRM, whereas differentiated Caco-2 cells exhibited low adhesion of GO sheets. Transmission electron microscopy analysis revealed internalisation of both applied GO (small and large) by undifferentiated Caco-2 cells. Even large GO sheets with lateral dimensions up to 10 µm, were found internalised by undifferentiated cells, presumably by macropinocytosis. In contrast, no GO uptake could be found for differentiated Caco-2 cells exhibiting an enterocyte-like morphology with apical brush border. Our results show that the internalisation of GO is highly dependent on the cell differentiation status of human intestinal cells. During differentiation Caco-2 cells undergo intense phenotypic changes which lead to a dramatic decrease in GRM internalisation. The results support the hypothesis that the cell surface topography of differentiated Caco-2 cells given by the brush border leads to low adhesion of GO sheets and sterical hindrance for material uptake. In addition, the mechanical properties of GRM, especially flexibility of the sheets, seem to be an important factor for

  7. Quantitative label-free proteomics for discovery of biomarkers in cerebrospinal fluid: assessment of technical and inter-individual variation.

    Directory of Open Access Journals (Sweden)

    Richard J Perrin

    Full Text Available Biomarkers are required for pre-symptomatic diagnosis, treatment, and monitoring of neurodegenerative diseases such as Alzheimer's disease. Cerebrospinal fluid (CSF is a favored source because its proteome reflects the composition of the brain. Ideal biomarkers have low technical and inter-individual variability (subject variance among control subjects to minimize overlaps between clinical groups. This study evaluates a process of multi-affinity fractionation (MAF and quantitative label-free liquid chromatography tandem mass spectrometry (LC-MS/MS for CSF biomarker discovery by (1 identifying reparable sources of technical variability, (2 assessing subject variance and residual technical variability for numerous CSF proteins, and (3 testing its ability to segregate samples on the basis of desired biomarker characteristics.Fourteen aliquots of pooled CSF and two aliquots from six cognitively normal individuals were randomized, enriched for low-abundance proteins by MAF, digested endoproteolytically, randomized again, and analyzed by nano-LC-MS. Nano-LC-MS data were time and m/z aligned across samples for relative peptide quantification. Among 11,433 aligned charge groups, 1360 relatively abundant ones were annotated by MS2, yielding 823 unique peptides. Analyses, including Pearson correlations of annotated LC-MS ion chromatograms, performed for all pairwise sample comparisons, identified several sources of technical variability: i incomplete MAF and keratins; ii globally- or segmentally-decreased ion current in isolated LC-MS analyses; and iii oxidized methionine-containing peptides. Exclusion of these sources yielded 609 peptides representing 81 proteins. Most of these proteins showed very low coefficients of variation (CV<5% whether they were quantified from the mean of all or only the 2 most-abundant peptides. Unsupervised clustering, using only 24 proteins selected for high subject variance, yielded perfect segregation of pooled and

  8. Label-free quantitative proteomics to investigate strawberry fruit proteome changes under controlled atmosphere and low temperature storage.

    Science.gov (United States)

    Li, Li; Luo, Zisheng; Huang, Xinhong; Zhang, Lu; Zhao, Pengyu; Ma, Hongyuan; Li, Xihong; Ban, Zhaojun; Liu, Xia

    2015-04-29

    To elucidate the mechanisms contributing to fruit responses to senescence and stressful environmental stimuli under low temperature (LT) and controlled atmosphere (CA) storage, a label-free quantitative proteomic investigation was conducted in strawberry (Fragaria ananassa, Duch. cv. 'Akihime'). Postharvest physiological quality traits including firmness, total soluble solids, total acidity, ascorbic acid and volatile production were characterized following storage under different conditions. The observed post-storage protein expression profiles may be associated with delayed senescence features in strawberry. A total of 454 proteins were identified in differentially treated strawberry fruits. Quantitative analysis, using normalized spectral counts, revealed 73 proteins common to all treatments, which formed three clusters in a hierarchical clustering analysis. The proteins spanned a range of functions in various metabolic pathways and networks involved in carbohydrate and energy metabolism, volatile biosynthesis, phenylpropanoid activity, stress response and protein synthesis, degradation and folding. After CA and LT storage, 16 (13) and 11 (17) proteins, respectively, were significantly increased (decreased) in abundance, while expression profile of 12 proteins was significantly changed by both CA and LT. To summarize, the differential variability of abundance in strawberry proteome, working in a cooperative manner, provided an overview of the biological processes that occurred during CA and LT storage. Controlled atmosphere storage at an optimal temperature is regarded to be an effective postharvest technology to delay fruit senescence and maintain fruit quality during shelf life. Nonetheless, little information on fruit proteomic changes under controlled atmosphere and/or low temperature storage is available. The significance of this paper is that it is the first study employing a label-free approach in the investigation of strawberry fruit response to

  9. Rapid and Label-Free Strategy to Isolate Aptamers for Metal Ions.

    Science.gov (United States)

    Qu, Hao; Csordas, Andrew T; Wang, Jinpeng; Oh, Seung Soo; Eisenstein, Michael S; Soh, Hyongsok Tom

    2016-08-23

    Generating aptamers that bind to specific metal ions is challenging because existing aptamer discovery methods typically require chemical labels or modifications that can alter the structure and properties of the ions. In this work, we report an aptamer discovery method that enables us to generate high-quality structure-switching aptamers (SSAs) that undergo a conformational change upon binding a metal ion target, without the requirement of labels or chemical modifications. Our method is more efficient than conventional selection methods because it enables direct measurement of target binding via fluorescence-activated cell sorting (FACS), isolating only the desired aptamers with the highest affinity. Using this strategy, we obtained a highly specific DNA SSA with ∼30-fold higher affinity than the best aptamer for Hg(2+) in the literature. We also discovered DNA aptamers that bind to Cu(2+) with excellent affinity and specificity. Both aptamers were obtained within four rounds of screening, demonstrating the efficiency of our aptamer discovery method. Given the growing availability of FACS, we believe our method offers a general strategy for discovering high-quality aptamers for other ions and small-molecule targets in an efficient and reproducible manner.

  10. 75 FR 78980 - Application to Export Electric Energy; Direct Energy Marketing, Inc.

    Science.gov (United States)

    2010-12-17

    ... Marketing, Inc. AGENCY: Office of Electricity Delivery and Energy Reliability, DOE. ACTION: Notice of Application. SUMMARY: Direct Energy Marketing, Inc. (DEMI) has applied to renew its authority to transmit..., Federal power marketing agencies, and other entities within the United States. The existing international...

  11. Directed Orientation of Asymmetric Composite Dumbbells by Electric Field Induced Assembly

    NARCIS (Netherlands)

    Nagao, D.; Sugimoto, M.; Okada, A.; Ishii, H.; Konno, M.; Imhof, A.; van Blaaderen, A.

    2012-01-01

    Assembly and directed orientation of anisotropic particles with an external ac electric field in a range from 1 kHz to 2 MHz were studied for asymmetric composite dumbbells incorporating a silica, titania, or titania/silica (titania:silica = 75:25 vol %) sphere. The asymmetric composite dumbbells,

  12. Electric power generating plant having direct coupled steam and compressed air cycles

    Science.gov (United States)

    Drost, Monte K.

    1982-01-01

    An electric power generating plant is provided with a Compressed Air Energy Storage (CAES) system which is directly coupled to the steam cycle of the generating plant. The CAES system is charged by the steam boiler during off peak hours, and drives a separate generator during peak load hours. The steam boiler load is thereby levelized throughout an operating day.

  13. Electric power generating plant having direct-coupled steam and compressed-air cycles

    Science.gov (United States)

    Drost, M.K.

    1981-01-07

    An electric power generating plant is provided with a Compressed Air Energy Storage (CAES) system which is directly coupled to the steam cycle of the generating plant. The CAES system is charged by the steam boiler during off peak hours, and drives a separate generator during peak load hours. The steam boiler load is thereby levelized throughout an operating day.

  14. 76 FR 68634 - Airworthiness Directives; General Electric Company (GE) CF6 Turbofan Engines

    Science.gov (United States)

    2011-11-07

    ... Airworthiness Directives; General Electric Company (GE) CF6 Turbofan Engines AGENCY: Federal Aviation... (AD) for the engines identified above. That AD currently requires initial and repetitive visual inspections of the forward engine mount assembly side links for cracks, stripping and reapplying the Sermetel...

  15. Optical elastic scattering for early label-free identification of clinical pathogens

    Science.gov (United States)

    Genuer, Valentin; Gal, Olivier; Méteau, Jérémy; Marcoux, Pierre; Schultz, Emmanuelle; Lacot, Éric; Maurin, Max; Dinten, Jean-Marc

    2016-03-01

    We report here on the ability of elastic light scattering in discriminating Gram+, Gram- and yeasts at an early stage of growth (6h). Our technique is non-invasive, low cost and does require neither skilled operators nor reagents. Therefore it is compatible with automation. It is based on the analysis of the scattering pattern (scatterogram) generated by a bacterial microcolony growing on agar, when placed in the path of a laser beam. Measurements are directly performed on closed Petri dishes. The characteristic features of a given scatterogram are first computed by projecting the pattern onto the Zernike orthogonal basis. Then the obtained data are compared to a database so that machine learning can yield identification result. A 10-fold cross-validation was performed on a database over 8 species (15 strains, 1906 scatterograms), at 6h of incubation. It yielded a 94% correct classification rate between Gram+, Gram- and yeasts. Results can be improved by using a more relevant function basis for projections, such as Fourier-Bessel functions. A fully integrated instrument has been installed at the Grenoble hospital's laboratory of bacteriology and a validation campaign has been started for the early screening of MSSA and MRSA (Staphylococcus aureus, methicillin-resistant S. aureus) carriers. Up to now, all the published studies about elastic scattering were performed in a forward mode, which is restricted to transparent media. However, in clinical diagnostics, most of media are opaque, such as blood-supplemented agar. That is why we propose a novel scheme capable of collecting back-scattered light which provides comparable results.

  16. Protein profiling in hepatocellular carcinoma by label-free quantitative proteomics in two west African populations.

    Directory of Open Access Journals (Sweden)

    Haddy K S Fye

    Full Text Available Hepatocellular Carcinoma is the third most common cause of cancer related death worldwide, often diagnosed by measuring serum AFP; a poor performance stand-alone biomarker. With the aim of improving on this, our study focuses on plasma proteins identified by Mass Spectrometry in order to investigate and validate differences seen in the respective proteomes of controls and subjects with LC and HCC.Mass Spectrometry analysis using liquid chromatography electro spray ionization quadrupole time-of-flight was conducted on 339 subjects using a pooled expression profiling approach. ELISA assays were performed on four significantly differentially expressed proteins to validate their expression profiles in subjects from the Gambia and a pilot group from Nigeria. Results from this were collated for statistical multiplexing using logistic regression analysis.Twenty-six proteins were identified as differentially expressed between the three subject groups. Direct measurements of four; hemopexin, alpha-1-antitrypsin, apolipoprotein A1 and complement component 3 confirmed their change in abundance in LC and HCC versus control patients. These trends were independently replicated in the pilot validation subjects from Nigeria. The statistical multiplexing of these proteins demonstrated performance comparable to or greater than ALT in identifying liver cirrhosis or carcinogenesis. This exercise also proposed preliminary cut offs with achievable sensitivity, specificity and AUC statistics greater than reported AFP averages.The validated changes of expression in these proteins have the potential for development into high-performance tests usable in the diagnosis and or monitoring of HCC and LC patients. The identification of sustained expression trends strengthens the suggestion of these four proteins as worthy candidates for further investigation in the context of liver disease. The statistical combinations also provide a novel inroad of analyses able to propose

  17. Concurrent electrical cervicomedullary stimulation and cervical transcutaneous spinal direct current stimulation result in a stimulus interaction.

    Science.gov (United States)

    Dongés, Siobhan C; Bai, Siwei; Taylor, Janet L

    2017-10-01

    What is the central question of this study? We previously showed that the motor pathway is not modified after cervical transcutaneous spinal direct current stimulation (tsDCS) applied using anterior-posterior electrodes. Here, we examine the motor pathway during stimulation. What is the main finding and its importance? We show that electrically elicited muscle responses to cervicomedullary stimulation are modified during tsDCS, whereas magnetically elicited responses are not. Modelling reveals electrical field modifications during concurrent tsDCS and electrical cervicomedullary stimulation. Changes in muscle response probably result from electrical field modifications rather than physiological changes. Care should be taken when applying electrical stimuli simultaneously. Transcutaneous spinal direct current stimulation (tsDCS) can modulate neuronal excitability within the human spinal cord; however, few studies have used tsDCS at a cervical level. This study aimed to characterize cervical tsDCS further by observing its acute effects on motor responses to transcranial magnetic stimulation and cervicomedullary stimulation. In both studies 1 and 2, participants (study 1, n = 8, four female; and study 2, n = 8, three female) received two periods of 10 min, 3 mA cervical tsDCS on the same day through electrodes placed in an anterior-posterior configuration over the neck; one period with the cathode posterior (c-tsDCS) and the other with the anode posterior (a-tsDCS). In study 1, electrically elicited cervicomedullary motor evoked potentials (eCMEPs) and transcranial magnetic stimulation-elicited motor evoked potentials (MEPs) were measured in biceps brachii and flexor carpi radialis before, during and after each tsDCS period. In study 2, eCMEPs and magnetically elicited CMEPs (mCMEPs) were measured before, during and after each tsDCS period. For study 3, computational modelling was used to observe possible interactions of cervical tsDCS and electrical

  18. Label-free silicon nanodots featured ratiometric fluorescent aptasensor for lysosomal imaging and pH measurement.

    Science.gov (United States)

    Zhang, Yanan; Guo, Shan; Cheng, Shibo; Ji, Xinghu; He, Zhike

    2017-08-15

    The homeostasis of lysosomal pH is crucial in cell physiology. Developing small fluorescent nanosensors for lysosome imaging and ratiometric measurement of pH is highly demanded yet challenging. Herein, a pH-sensitive fluorescein tagged aptamer AS1411 has been utilized to covalently modify the label-free fluorescent silicon nanodots via a crosslinker for construction of a ratiometric pH biosensor. The established aptasensor exhibits the advantages of ultrasmall size, hypotoxicity, excellent pH reversibility and good photostability, which favors its application in an intracellular environment. Using human breast MCF-7 cancer cells and MCF-10A normal cells as the model, this aptasensor shows cell specificity for cancer cells and displays a wide pH response range of 4.5-8.0 in living cells. The results demonstrate that the pH of MCF-7 cells is 5.1, which is the expected value for acidic organelles. Lysosome imaging and accurate measurement of pH in MCF-7 cells have been successfully conducted based on this nanosensor via fluorescent microscopy and flow cytometry. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. A label-free detector for liquid chromatography systems using mm-wave technology: First proof of concept.

    Science.gov (United States)

    Zhang, Y; Declerck, S; Mangelings, D; He, G; Matvejev, V; Vander Heyden, Y; Stiens, J

    2017-09-22

    The development of millimeter-wave (mm-wave) technology has enabled the study of bio-molecular interactions by means of electromagnetic waves with frequencies between 30 and 300GHz. In this study, an attempt has been made to exploit the possibility of mm-wave technology as alternative detection technique for liquid chromatographic (LC) systems. The goal is to design and fabricate a label-free mm-wave detector that is compatible with LC systems. As proof-of-concept experiments, the UV absorbing compounds praziquantel and trans-stilbene-oxide as well as a non-UV absorbing compound sorbitol are injected in an open capillary as well as a capillary with stationary phase and measured by both mm-wave and UV detectors. The in-house developed mm-wave detector is capable of detecting all compounds without the need for labelling. Although the detection limit of such detector still needs to be verified and occasionally improved in the future, it already shows great potential as an additional detection technique for LC systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Whole-cell biosensor for label-free detection of GPCR-mediated drug responses in personal cell lines.

    Science.gov (United States)

    Hillger, Julia M; Schoop, Jeffison; Boomsma, Dorret I; Slagboom, P Eline; IJzerman, Adriaan P; Heitman, Laura H

    2015-12-15

    Deciphering how genetic variation in drug targets such as G protein-coupled receptors (GPCRs) affects drug response is essential for precision medicine. GPCR signaling is traditionally investigated in artificial cell lines which do not provide sufficient physiological context. Patient-derived cell lines such as lymphoblastoid cell lines (LCLs) could represent the ideal cellular model system. Here we describe a novel label-free, whole-cell biosensor method for characterizing GPCR-mediated drug responses in LCLs. Generally, such biosensor technology is deemed only compatible with adherent cell lines. We optimized and applied the methodology to study cellular adhesion properties as well as GPCR drug responses in LCLs, which are suspension cells. Coating the detector surface with the extracellular matrix protein fibronectin resulted in cell adherence and allowed detection of cellular responses. A prototypical GPCR present on these cells, i.e. the cannabinoid receptor 2 (CB2), was selected for pharmacological characterization. Receptor activation with the agonist JWH133, blockade by antagonist AM630 as well as downstream signaling inhibition by PTX could be monitored sensitively and receptor-specifically. Potencies and effects were comparable between LCLs of two genetically unrelated individuals, providing the proof-of-principle that this biosensor technology can be applied to LCLs, despite their suspension cell nature, in order to serve as an in vitro model system for the evaluation of individual genetic influences on GPCR-mediated drug responses. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Fiber-optic Fourier transform infrared spectroscopy for remote label-free sensing of medical device surface contamination

    Science.gov (United States)

    Hassan, Moinuddin; Tan, Xin; Welle, Elissa; Ilev, Ilko

    2013-05-01

    As a potential major source of biochemical contamination, medical device surfaces are of critical safety concerns in the clinical practice and public health. The development of innovative sensing methods for accurate and real-time detection of medical device surface contamination is essential to protect patients from high risk infection. In this paper, we demonstrate an alternative fiber-optic Fourier Transform Infrared (FTIR) spectroscopy based sensing approach for remote, non-contact, and label-free detection of biochemical contaminants in the mid-infrared (mid-IR) region. The sensing probe is designed using mid-IR hollow fibers and FTIR measurements are carried out in reflection mode. Bovine Serum Albumin (BSA) and bacterial endotoxin of different concentrations under thoroughly dry condition are used to evaluate the detection sensitivity. The devised system can identify ≤0.0025% (≤4 × 1011 molecules) BSA and 0.5% (0.5 EU/ml) endotoxin concentration. The developed sensing approach may be applied to detect various pathogens that pose public health threats.

  2. 4-Fluoro-3-nitrophenyl grafted gold electrode based platform for label free electrochemical detection of interleukin-2 protein.

    Science.gov (United States)

    Arya, Sunil K; Park, Mi Kyoung

    2014-11-15

    A new platform based on 4-Fluoro-3-nitrophenyl (FNP) grafted gold disk electrode prepared via electrochemical reduction of 4-fluoro-3-nitrobenzene diazonium ion has been developed and utilized for biosensor fabrication. Anti-interleukin-2 (anti-IL2) antibody has been covalently immobilized onto FNP/Au surface and utilized for label free electrochemical impedance based detection of cytokine IL2. FNP acts as a bridge (cross-linker) between gold surface and anti-IL2, where fluoro group of FNP undergoes nucleophilic substitution by amino group of biomolecule and results in its covalent immobilization. The immobilization process and fabricated electrode have been characterized using contact angle (CA) measurements, cyclic voltammetry (CV) and electrochemical impedance (EIS) technique. CV studies show that FNP grafted surface provides conductive surface for anti-IL2 immobilization. The EIS response of studies as a function of IL2 concentrations exhibits a detection in linear range from 1 pg ml(-1) to 10 ng ml(-1) with minimum detectable concentration of 1 pg ml(-1). The electrode has been found to be selective against other cytokine molecules. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Label-free sensor for automatic identification of erythrocytes using digital in-line holographic microscopy and machine learning.

    Science.gov (United States)

    Go, Taesik; Byeon, Hyeokjun; Lee, Sang Joon

    2018-04-30

    Cell types of erythrocytes should be identified because they are closely related to their functionality and viability. Conventional methods for classifying erythrocytes are time consuming and labor intensive. Therefore, an automatic and accurate erythrocyte classification system is indispensable in healthcare and biomedical fields. In this study, we proposed a new label-free sensor for automatic identification of erythrocyte cell types using a digital in-line holographic microscopy (DIHM) combined with machine learning algorithms. A total of 12 features, including information on intensity distributions, morphological descriptors, and optical focusing characteristics, is quantitatively obtained from numerically reconstructed holographic images. All individual features for discocytes, echinocytes, and spherocytes are statistically different. To improve the performance of cell type identification, we adopted several machine learning algorithms, such as decision tree model, support vector machine, linear discriminant classification, and k-nearest neighbor classification. With the aid of these machine learning algorithms, the extracted features are effectively utilized to distinguish erythrocytes. Among the four tested algorithms, the decision tree model exhibits the best identification performance for the training sets (n = 440, 98.18%) and test sets (n = 190, 97.37%). This proposed methodology, which smartly combined DIHM and machine learning, would be helpful for sensing abnormal erythrocytes and computer-aided diagnosis of hematological diseases in clinic. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Systematic Comparison of Label-Free, Metabolic Labeling, and Isobaric Chemical Labeling for Quantitative Proteomics on LTQ Orbitrap Velos

    Energy Technology Data Exchange (ETDEWEB)

    Li, Zhou [ORNL; Adams, Rachel M [ORNL; Chourey, Karuna [ORNL; Hurst, Gregory {Greg} B [ORNL; Hettich, Robert {Bob} L [ORNL; Pan, Chongle [ORNL

    2012-01-01

    A variety of quantitative proteomics methods have been developed, including label-free, metabolic labeling, and isobaric chemical labeling using iTRAQ or TMT. Here, these methods were compared in terms of the depth of proteome coverage, quantification accuracy, precision, and reproducibility using a high-performance hybrid mass spectrometer, LTQ Orbitrap Velos. Our results show that (1) the spectral counting method provides the deepest proteome coverage for identification, but its quantification performance is worse than labeling-based approaches, especially the quantification reproducibility; (2) metabolic labeling and isobaric chemical labeling are capable of accurate, precise, and reproducible quantification and provide deep proteome coverage for quantification. Isobaric chemical labeling surpasses metabolic labeling in terms of quantification precision and reproducibility; (3) iTRAQ and TMT perform similarly in all aspects compared in the current study using a CID-HCD dual scan configuration. Based on the unique advantages of each method, we provide guidance for selection of the appropriate method for a quantitative proteomics study.

  5. Accounting for the Multiple Natures of Missing Values in Label-Free Quantitative Proteomics Data Sets to Compare Imputation Strategies.

    Science.gov (United States)

    Lazar, Cosmin; Gatto, Laurent; Ferro, Myriam; Bruley, Christophe; Burger, Thomas

    2016-04-01

    Missing values are a genuine issue in label-free quantitative proteomics. Recent works have surveyed the different statistical methods to conduct imputation and have compared them on real or simulated data sets and recommended a list of missing value imputation methods for proteomics application. Although insightful, these comparisons do not account for two important facts: (i) depending on the proteomics data set, the missingness mechanism may be of different natures and (ii) each imputation method is devoted to a specific type of missingness mechanism. As a result, we believe that the question at stake is not to find the most accurate imputation method in general but instead the most appropriate one. We describe a series of comparisons that support our views: For instance, we show that a supposedly "under-performing" method (i.e., giving baseline average results), if applied at the "appropriate" time in the data-processing pipeline (before or after peptide aggregation) on a data set with the "appropriate" nature of missing values, can outperform a blindly applied, supposedly "better-performing" method (i.e., the reference method from the state-of-the-art). This leads us to formulate few practical guidelines regarding the choice and the application of an imputation method in a proteomics context.

  6. Label-Free Quantitative Proteomic Analysis of Puccinia psidii Uredospores Reveals Differences of Fungal Populations Infecting Eucalyptus and Guava.

    Directory of Open Access Journals (Sweden)

    Maria Carolina Quecine

    Full Text Available Puccinia psidii sensu lato (s.l. is the causal agent of eucalyptus and guava rust, but it also attacks a wide range of plant species from the myrtle family, resulting in a significant genetic and physiological variability among populations accessed from different hosts. The uredospores are crucial to P. psidii dissemination in the field. Although they are important for the fungal pathogenesis, their molecular characterization has been poorly studied. In this work, we report the first in-depth proteomic analysis of P. psidii s.l. uredospores from two contrasting populations: guava fruits (PpGuava and eucalyptus leaves (PpEucalyptus. NanoUPLC-MSE was used to generate peptide spectra that were matched to the UniProt Puccinia genera sequences (UniProt database resulting in the first proteomic analysis of the phytopathogenic fungus P. psidii. Three hundred and fourty proteins were detected and quantified using Label free proteomics. A significant number of unique proteins were found for each sample, others were significantly more or less abundant, according to the fungal populations. In PpGuava population, many proteins correlated with fungal virulence, such as malate dehydrogenase, proteossomes subunits, enolases and others were increased. On the other hand, PpEucalyptus proteins involved in biogenesis, protein folding and translocation were increased, supporting the physiological variability of the fungal populations according to their protein reservoirs and specific host interaction strategies.

  7. A Label-Free Impedimetric DNA Sensor Based on a Nanoporous SnO2 Film: Fabrication and Detection Performance

    Directory of Open Access Journals (Sweden)

    Minh Hai Le

    2015-05-01

    Full Text Available Nanoporous SnO2 thin films were elaborated to serve as sensing electrodes for label-free DNA detection using electrochemical impedance spectroscopy (EIS. Films were deposited by an electrodeposition process (EDP. Then the non-Faradic EIS behaviour was thoroughly investigated during some different steps of functionalization up to DNA hybridization. The results have shown a systematic decrease of the impedance upon DNA hybridization. The impedance decrease is attributed to an enhanced penetration of ionic species within the film volume. Besides, the comparison of impedance variations upon DNA hybridization between the liquid and vapour phase processes for organosilane (APTES grafting on the nanoporous SnO2 films showed that vapour-phase method is more efficient. This is due to the fact that the vapour is more effective than the solution in penetrating the nanopores of the films. As a result, the DNA sensors built from vapour-treated silane layer exhibit a higher sensitivity than those produced from liquid-treated silane, in the range of tested target DNA concentration going to 10 nM. Finally, the impedance and fluorescence response signals strongly depend on the types of target DNA molecules, demonstrating a high selectivity of the process on nanoporous SnO2 films.

  8. Label-free pharmacological profiling based on dynamic mass redistribution for characterization and authentication of hazardous natural products.

    Science.gov (United States)

    Song, Hui-Peng; Wang, Hong; Zhao, Xiaoai; He, Ling; Zhong, Huailing; Wu, Si-Qi; Li, Ping; Yang, Hua

    2017-07-05

    Natural products are becoming increasingly popular in multiple fields involving medicines, foods and beverages. However, due to the frequent occurrence of poisoning incidents, their toxicity and safety have caused a serious concern. Here we report a method of biosensor-based two-phase pharmacological profiling (BTPP) for discovery, monitor and control of receptor-targeted natural products. BTPP uses a resonant waveguide grating biosensor for label-free and non-invasive detection of intracellular dynamic mass redistribution (DMR), a phenomenon caused by protein relocalization after receptors receiving stimulation from toxicants. The method can not only facilitate the identification of hazardous materials but also quantify their bioactivity by EC 50 . As a proof of concept, the method was successfully applied to recognize Daturae Flos (DF), an herb that can antagonize muscarinic acetylcholine M 2 receptor and further cause poisoning, from other easily confused species. BTPP combined with high performance liquid chromatography revealed that scopolamine and hyoscyamine in DF were the key marker compounds. Moreover, the method accurately picked out 2 M 2 receptor antagonists from 25 natural compounds, displaying its potential in high-throughput screening. This study provides a systematic illustration about the establishment, applicability and advantages of BTPP, which contributes to the safety assessment of natural products in related fields. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. A novel label-free microfluidic paper-based immunosensor for highly sensitive electrochemical detection of carcinoembryonic antigen.

    Science.gov (United States)

    Wang, Yang; Xu, Huiren; Luo, Jinping; Liu, Juntao; Wang, Li; Fan, Yan; Yan, Shi; Yang, Yue; Cai, Xinxia

    2016-09-15

    In this work, a highly sensitive label-free paper-based electrochemical immunosensor employing screen-printed working electrode (SPWE) for detection of carcinoembryonic antigen (CEA) was fabricated. In order to raise the detection sensitivity and immobilize anti-CEA, amino functional graphene (NH2-G)/thionine (Thi)/gold nanoparticles (AuNPs) nanocomposites were synthesized and coated on SPWE. The principle of the immunosensor determination was based on the fact that the decreased response currents of Thi were proportional to the concentrations of corresponding antigens due to the formation of antibody-antigen immunocomplex. Experimental results revealed that the immunoassay enabled the determination of standard CEA solutions with linear working ranges of 50pgmL(-1) to 500ngmL(-1), the limit of detections for CEA is 10pgmL(-1) (S/N=3) and its corresponding correlation coefficients were 0.996. Furthermore, the proposed immunosensor could be used for the determination of clinical serum samples. A large number of clinical serum samples were detected and the relative errors between measured values and reference concentrations were calculated. Results showed that this novel paper-based electrochemical immunosensor could provide a new platform for low cost, sensitive, specific, and point-of-care diagnosis in cancer detection. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Label-Free Quantitative Proteomic Analysis of Puccinia psidii Uredospores Reveals Differences of Fungal Populations Infecting Eucalyptus and Guava

    Science.gov (United States)

    Bini, Andressa Peres; Regiani, Thais; Franceschini, Lívia Maria; Budzinski, Ilara Gabriela Frasson; Marques, Felipe Garbelini; Labate, Mônica Teresa Veneziano; Guidetti-Gonzalez, Simone; Moon, David Henry; Labate, Carlos Alberto

    2016-01-01

    Puccinia psidii sensu lato (s.l.) is the causal agent of eucalyptus and guava rust, but it also attacks a wide range of plant species from the myrtle family, resulting in a significant genetic and physiological variability among populations accessed from different hosts. The uredospores are crucial to P. psidii dissemination in the field. Although they are important for the fungal pathogenesis, their molecular characterization has been poorly studied. In this work, we report the first in-depth proteomic analysis of P. psidii s.l. uredospores from two contrasting populations: guava fruits (PpGuava) and eucalyptus leaves (PpEucalyptus). NanoUPLC-MSE was used to generate peptide spectra that were matched to the UniProt Puccinia genera sequences (UniProt database) resulting in the first proteomic analysis of the phytopathogenic fungus P. psidii. Three hundred and fourty proteins were detected and quantified using Label free proteomics. A significant number of unique proteins were found for each sample, others were significantly more or less abundant, according to the fungal populations. In PpGuava population, many proteins correlated with fungal virulence, such as malate dehydrogenase, proteossomes subunits, enolases and others were increased. On the other hand, PpEucalyptus proteins involved in biogenesis, protein folding and translocation were increased, supporting the physiological variability of the fungal populations according to their protein reservoirs and specific host interaction strategies. PMID:26731728

  11. Label-free fluorescence strategy for sensitive detection of adenosine triphosphate using a loop DNA probe with low background noise.

    Science.gov (United States)

    Lin, Chunshui; Cai, Zhixiong; Wang, Yiru; Zhu, Zhi; Yang, Chaoyong James; Chen, Xi

    2014-07-15

    A simple, rapid, label-free, and ultrasensitive fluorescence strategy for adenosine triphosphate (ATP) detection was developed using a loop DNA probe with low background noise. In this strategy, a loop DNA probe, which is the substrate for both ligation and digestion enzyme reaction, was designed. SYBR green I (SG I), a double-stranded specific dye, was applied for the readout fluorescence signal. Exonuclease I (Exo I) and exonuclease III (Exo III), sequence-independent nucleases, were selected to digest the loop DNA probe in order to minimize the background fluorescence signal. As a result, in the absence of ATP, the loop DNA was completely digested by Exo I and Exo III, leading to low background fluorescence owing to the weak electrostatic interaction between SG I and mononucleotides. On the other hand, ATP induced the ligation of the nicking site, and the sealed loop DNA resisted the digestion of Exo I and ExoIII, resulting in a remarkable increase of fluorescence response. Upon background noise reduction, the sensitivity of the ATP determination was improved significantly, and the detection limitation was found to be 1.2 pM, which is much lower than that in almost all the previously reported methods. This strategy has promise for wide application in the determination of ATP.

  12. Biotin-Streptavidin Binding Interactions of Dielectric Filled Silicon Bulk Acoustic Resonators for Smart Label-Free Biochemical Sensor Applications

    Directory of Open Access Journals (Sweden)

    Amir Heidari

    2014-03-01

    Full Text Available Sensor performance of a dielectric filled silicon bulk acoustic resonator type label-free biosensor is verified with biotin-streptavidin binding interactions as a model system. The mass sensor is a micromachined silicon square plate with a dielectric filled capacitive excitation mechanism. The resonance frequency of the biotin modified resonator decreased 315 ppm when exposed to streptavidin solution for 15 min with a concentration of 10−7 M, corresponding to an added mass of 3.43 ng on the resonator surface. An additional control is added by exposing a bovine serum albumin (BSA-covered device to streptavidin in the absence of the attached biotin. No resonance frequency shift was observed in the control experiment, which confirms the specificity of the detection. The sensor-to-sensor variability is also measured to be 4.3%. Consequently, the developed sensor can be used to observe in biotin-streptavidin interaction without the use of labelling or molecular tags. In addition, biosensor can be used in a variety of different immunoassay tests.

  13. Label-free quantitative mass spectrometry for analysis of protein antigens in a meningococcal group B outer membrane vesicle vaccine.

    Science.gov (United States)

    Dick, Lawrence W; Mehl, John T; Loughney, John W; Mach, Anna; Rustandi, Richard R; Ha, Sha; Zhang, Lan; Przysiecki, Craig T; Dieter, Lance; Hoang, Van M

    2015-01-01

    The development of a multivalent outer membrane vesicle (OMV) vaccine where each strain contributes multiple key protein antigens presents numerous analytical challenges. One major difficulty is the ability to accurately and specifically quantitate each antigen, especially during early development and process optimization when immunoreagents are limited or unavailable. To overcome this problem, quantitative mass spectrometry methods can be used. In place of traditional mass assays such as enzyme-linked immunosorbent assays (ELISAs), quantitative LC-MS/MS using multiple reaction monitoring (MRM) can be used during early-phase process development to measure key protein components in complex vaccines in the absence of specific immunoreagents. Multiplexed, label-free quantitative mass spectrometry methods using protein extraction by either detergent or 2-phase solvent were developed to quantitate levels of several meningococcal serogroup B protein antigens in an OMV vaccine candidate. Precision was demonstrated to be less than 15% RSD for the 2-phase extraction and less than 10% RSD for the detergent extraction method. Accuracy was 70 to 130% for the method using a 2-phase extraction and 90-110% for detergent extraction. The viability of MS-based protein quantification as a vaccine characterization method was demonstrated and advantages over traditional quantitative methods were evaluated. Implementation of these MS-based quantification methods can help to decrease the development time for complex vaccines and can provide orthogonal confirmation of results from existing antigen quantification techniques.

  14. Summarization vs Peptide-Based Models in Label-Free Quantitative Proteomics: Performance, Pitfalls, and Data Analysis Guidelines.

    Science.gov (United States)

    Goeminne, Ludger J E; Argentini, Andrea; Martens, Lennart; Clement, Lieven

    2015-06-05

    Quantitative label-free mass spectrometry is increasingly used to analyze the proteomes of complex biological samples. However, the choice of appropriate data analysis methods remains a major challenge. We therefore provide a rigorous comparison between peptide-based models and peptide-summarization-based pipelines. We show that peptide-based models outperform summarization-based pipelines in terms of sensitivity, specificity, accuracy, and precision. We also demonstrate that the predefined FDR cutoffs for the detection of differentially regulated proteins can become problematic when differentially expressed (DE) proteins are highly abundant in one or more samples. Care should therefore be taken when data are interpreted from samples with spiked-in internal controls and from samples that contain a few very highly abundant proteins. We do, however, show that specific diagnostic plots can be used for assessing differentially expressed proteins and the overall quality of the obtained fold change estimates. Finally, our study also illustrates that imputation under the "missing by low abundance" assumption is beneficial for the detection of differential expression in proteins with low abundance, but it negatively affects moderately to highly abundant proteins. Hence, imputation strategies that are commonly implemented in standard proteomics software should be used with care.

  15. Real-time Image Processing for Microscopy-based Label-free Imaging Flow Cytometry in a Microfluidic Chip.

    Science.gov (United States)

    Heo, Young Jin; Lee, Donghyeon; Kang, Junsu; Lee, Keondo; Chung, Wan Kyun

    2017-09-14

    Imaging flow cytometry (IFC) is an emerging technology that acquires single-cell images at high-throughput for analysis of a cell population. Rich information that comes from high sensitivity and spatial resolution of a single-cell microscopic image is beneficial for single-cell analysis in various biological applications. In this paper, we present a fast image-processing pipeline (R-MOD: Real-time Moving Object Detector) based on deep learning for high-throughput microscopy-based label-free IFC in a microfluidic chip. The R-MOD pipeline acquires all single-cell images of cells in flow, and identifies the acquired images as a real-time process with minimum hardware that consists of a microscope and a high-speed camera. Experiments show that R-MOD has the fast and reliable accuracy (500 fps and 93.3% mAP), and is expected to be used as a powerful tool for biomedical and clinical applications.

  16. Organic Liquids-Responsive β-Cyclodextrin-Functionalized Graphene-Based Fluorescence Probe: Label-Free Selective Detection of Tetrahydrofuran

    Directory of Open Access Journals (Sweden)

    Huawen Hu

    2014-06-01

    Full Text Available In this study, a label-free graphene-based fluorescence probe used for detection of volatile organic liquids was fabricated by a simple, efficient and low-cost method. To fabricate the probe, a bio-based β-cyclodextrin (β-CD was firstly grafted on reduced graphene surfaces effectively and uniformly, as evidenced by various characterization techniques such as Ultraviolet/Visible spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, scanning electron microscopy and transmission electron microscopy. The subsequent inclusion of Rhodamine B (RhB into the inner cavities of the β-CD grafted on the graphene surfaces was achieved easily by a solution mixing method, which yielded the graphene-based fluorescent switch-on probe. In addition, the gradual and controllable quenching of RhB by Fluorescence Resonance Energy Transfer from RhB to graphene during the process of stepwise accommodation of the RhB molecules into the β-CD-functionalized graphene was investigated in depth. A wide range of organic solvents was examined using the as-fabricated fluorescence probe, which revealed the highest sensitivity to tetrahydrofuran with the detection limit of about 1.7 μg/mL. Some insight into the mechanism of the different responsive behaviors of the fluorescence sensor to the examined targets was also described.

  17. Application on Gold Nanoparticles-Dotted 4-Nitrophenylazo Graphene in a Label-Free Impedimetric Deoxynivalenol Immunosensor

    Directory of Open Access Journals (Sweden)

    Christopher Edozie Sunday

    2015-02-01

    Full Text Available In this paper, we report a new concept to construct a label-free electrochemical inhibition-based immunosensor for the detection of the mycotoxin deoxynivalenol (DON in cereal samples. The electrochemical impedance spectroscopy of tris(bipyridine ruthenium (II chloride was used as a marker enhanced with gold nanoparticles-dotted 4-nitrophenylazo functionalized graphene (AuNp/G/PhNO2 nanocatalyst mediated in Nafion on a glassy carbon electrode. Under the optimized conditions, the formation of immunocomplexes inhibited electron flow and increased the charge transfer resistance of the sensing interface linearly. The change in impedance was proportional to DON concentrations in the range of 6–30 ng/mL with a sensitivity and detection limit of 32.14 ΩL/ng and 0.3 µg/mL, respectively, which compares favorably with the ELISA result. The proposed sensor had a stability of 80.3%, good precision and selectivity in DON standard solution containing different interfering agents, indicating promising application prospect for this strategy in designing impedimetric, electrochemiluminescent, voltammetric or amperometric sensors.

  18. Electrochemical label-free and sensitive nanobiosensing of DNA hybridization by graphene oxide modified pencil graphite electrode.

    Science.gov (United States)

    Ahour, F; Shamsi, A

    2017-09-01

    Based on the strong interaction between single-stranded DNA (ss-DNA) and graphene material, we have constructed a novel label-free electrochemical biosensor for rapid and facile detection of short sequences ss-DNA molecules related to hepatitis C virus 1a using graphene oxide modified pencil graphite electrode. The sensing mechanism is based on the superior adsorption of single-stranded DNA to GO over double stranded DNA (ds-DNA). The intrinsic guanine oxidation signal measured by differential pulse voltammetry (DPV) has been used for duplex DNA formation detection. The probe ss-DNA adsorbs onto the surface of GO via the π- π* stacking interactions leading to a strong background guanine oxidation signal. In the presence of complementary target, formation of helix which has weak binding ability to GO induced ds-DNA to release from the electrode surface and significant variation in differential pulse voltammetric response of guanine bases. The results indicated that the oxidation peak current was proportional to the concentration of complementary strand in the range of 0.1 nM-0.5 μM with a detection limit of 4.3 × 10 -11  M. The simple fabricated electrochemical biosensor has high sensitivity, good selectivity, and could be applied as a new platform for a range of target molecules in future. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. High-throughput label-free screening of euglena gracilis with optofluidic time-stretch quantitative phase microscopy

    Science.gov (United States)

    Guo, Baoshan; Lei, Cheng; Ito, Takuro; Yaxiaer, Yalikun; Kobayashi, Hirofumi; Jiang, Yiyue; Tanaka, Yo; Ozeki, Yasuyuki; Goda, Keisuke

    2017-02-01

    The development of reliable, sustainable, and economical sources of alternative fuels is an important, but challenging goal for the world. As an alternative to liquid fossil fuels, microalgal biofuel is expected to play a key role in reducing the detrimental effects of global warming since microalgae absorb atmospheric CO2 via photosynthesis. Unfortunately, conventional analytical methods only provide population-averaged lipid contents and fail to characterize a diverse population of microalgal cells with single-cell resolution in a noninvasive and interference-free manner. Here we demonstrate high-throughput label-free single-cell screening of lipid-producing microalgal cells with optofluidic time-stretch quantitative phase microscopy. In particular, we use Euglena gracilis - an attractive microalgal species that produces wax esters (suitable for biodiesel and aviation fuel after refinement) within lipid droplets. Our optofluidic time-stretch quantitative phase microscope is based on an integration of a hydrodynamic-focusing microfluidic chip, an optical time-stretch phase-contrast microscope, and a digital image processor equipped with machine learning. As a result, it provides both the opacity and phase contents of every single cell at a high throughput of 10,000 cells/s. We characterize heterogeneous populations of E. gracilis cells under two different culture conditions to evaluate their lipid production efficiency. Our method holds promise as an effective analytical tool for microalgaebased biofuel production.

  20. Label-free electrochemical detection of botulinum neurotoxin type E based on its enzymatic activity using interdigitated electrodes

    Science.gov (United States)

    Hyun, Sang Hwa; Park, Dae Keun; Kang, Aeyeon; Kim, Soohyun; Kim, Daehee; Shin, Yu Mi; Song, Ji-Joon; Yun, Wan Soo

    2016-02-01

    We report a simple label-free electrochemical method of detecting low concentrations of botulinum neurotoxin type E light chain (BoNT/E LC) based on its peptide cleavage activity. Dual-mode cyclic voltammetry was employed to observe changes in the redox signal of ferri-/ferro-cyanide on interdigitated microelectrodes, whose surfaces were covered by peptides designed from synaptosomal-associated protein 25 to be cleaved by BoNT/E LC. With the introduction of BoNT/E LC, the redox signal showed a time-dependent increase due to cleavage of the immobilized peptide molecules. In addition to the increased redox signal intensity, its time-dependence can be considered as a strong evidence of BoNT/E sensing, since the time-dependent increase can only result from the enzymatic activity of BoNT/E LC. Using this method, BoNT/E LC, at concentrations as low as 5 pg/ml, was readily measurable with only an hour of incubation.

  1. Proteomic profile of dormancy within Staphylococcus epidermidis biofilms using iTRAQ and label-free strategies.

    Science.gov (United States)

    Carvalhais, Virginia; Cerca, Nuno; Vilanova, Manuel; Vitorino, Rui

    2015-03-01

    Staphylococcus epidermidis is an important nosocomial bacterium among carriers of indwelling medical devices, since it has a strong ability to form biofilms. The presence of dormant bacteria within a biofilm is one of the factors that contribute to biofilm antibiotic tolerance and immune evasion. Here, we provide a detailed characterization of the quantitative proteomic profile of S. epidermidis biofilms with different proportions of dormant bacteria. A total of 427 and 409 proteins were identified by label-free and label-based quantitative methodologies, respectively. From these, 29 proteins were found to be differentially expressed between S. epidermidis biofilms with prevented and induced dormancy. Proteins overexpressed in S. epidermidis with prevented dormancy were associated with ribosome synthesis pathway, which reflects the metabolic state of dormant bacteria. In the opposite, underexpressed proteins were related to catalytic activity and ion binding, with involvement in purine, arginine, and proline metabolism. Additionally, GTPase activity seems to be enhanced in S. epidermidis biofilm with induced dormancy. The role of magnesium in dormancy modulation was further investigated with bioinformatics tool based in predicted interactions. The main molecular function of proteins, which strongly interact with magnesium, was nucleic acid binding. Different proteomic strategies allowed to obtain similar results and evidenced that prevented dormancy led to an expression of a markedly different repertoire of proteins in comparison to the one of dormant biofilms.

  2. Label-free biomolecular characterization of human breast cancer tissue with stimulated Raman scattering (SRS) spectral imaging (Conference Presentation)

    Science.gov (United States)

    Lu, Fa-Ke F.; Calligaris, David; Suo, Yuanzhen; Santagata, Sandro; Golby, Alexandra J.; Xie, X. Sunney; Mallory, Melissa A.; Golshan, Mehra; Dillon, Deborah A.; Agar, Nathalie Y. R.

    2017-02-01

    Stimulated Raman scattering (SRS) microscopy has been used for rapid label-free imaging of various biomolecules and drugs in living cells and tissues (Science, doi:10.1126/science.aaa8870). Our recent work has demonstrated that lipid and protein mapping of cancer tissue renders pathology-like images, providing essential histopathological information with subcellular resolution of the entire specimen (Cancer Research, doi: 10.1158/0008-5472.CAN-16-027). We have also established the first SRS imaging Atlas of human brain tumors (Harvard Dataverse, doi: (doi:10.7910/DVN/EZW4EK). SRS imaging of tissue could provide invaluable information for cancer diagnosis and surgical guidance in two aspects: rapid surgical pathology and quantitative biomolecular characterization. In this work, we present the use of SRS microscopy for characterization of a few essential biomolecules in breast cancer. Human breast cancer tissue specimens at the tumor core, tumor margin and normal area (5 cm away from the tumor) from surgical cases will be imaged with SRS at multiple Raman shifts, including the peaks for lipid, protein, blood (absorption), collagen, microcalcification (calcium phosphates and calcium oxalate) and carotenoids. Most of these Raman shifts have relatively strong Raman cross sections, which ensures high-quality and fast imaging. This proof-of-principle study is sought to demonstrate the feasibility and potential of SRS imaging for ambient diagnosis and surgical guidance of breast cancer.

  3. Label-free detection and identification of waterborne parasites using a microfluidic multi-angle laser scattering system

    Science.gov (United States)

    Huang, Wei; Yang, Limei; Lei, Lei; Li, Feng

    2017-10-01

    A microfluidic-based multi-angle laser scattering (MALS) system capable of acquiring scattering patterns of a single particle is designed and demonstrated. The system includes a sheathless nozzle microfluidic glass chip, and an on-chip MALS unit being in alignment with the nozzle exit in the chip. The size and relative refractive indices (RI) of polystyrene (PS) microspheres were deduced with accuracies of 60 nm and 0.002 by comparing the experimental scattering patterns with theoretical ones. We measured scattering patterns of waterborne parasites i.e., Cryptosporidium parvum (C.parvum) and Giardia lamblia (G. lamblia), and some other representative species suspended in deionized water at a maximum flow rate of 12 μL/min, and a maximum of 3000 waterborne parasites can be identified within one minute with a mean accuracy higher than 96% by classification of distinctive scattering patterns using a support-vector-machine (SVM) algorithm. The system provides a promising tool for label-free detection of waterborne parasites and other biological contaminants.

  4. Single-walled carbon nanotubes based chemiresistive genosensor for label-free detection of human rheumatic heart disease

    International Nuclear Information System (INIS)

    Singh, Swati; Kumar, Ashok; Khare, Shashi; Mulchandani, Ashok; Rajesh

    2014-01-01

    A specific and ultrasensitive, label free single-walled carbon nanotubes (SWNTs) based chemiresistive genosensor was fabricated for the early detection of Streptococcus pyogenes infection in human causing rheumatic heart disease. The mga gene of S. pyogenes specific 24 mer ssDNA probe was covalently immobilized on SWNT through a molecular bilinker, 1-pyrenemethylamine, using carbodiimide coupling reaction. The sensor was characterized by the current-voltage (I-V) characteristic curve and scanning electron microscopy. The sensing performance of the sensor was studied with respect to changes in conductance in SWNT channel based on hybridization of the target S. pyogenes single stranded genomic DNA (ssG-DNA) to its complementary 24 mer ssDNA probe. The sensor shows negligible response to non-complementary Staphylococcus aureus ssG-DNA, confirming the specificity of the sensor only with S. pyogenes. The genosensor exhibited a linear response to S. pyogenes G-DNA from 1 to1000 ng ml −1 with a limit of detection of 0.16 ng ml −1

  5. Analysis of Drosophila melanogaster proteome dynamics during embryonic development by a combination of label-free proteomics approaches.

    Science.gov (United States)

    Fabre, Bertrand; Korona, Dagmara; Groen, Arnoud; Vowinckel, Jakob; Gatto, Laurent; Deery, Michael J; Ralser, Markus; Russell, Steven; Lilley, Kathryn S

    2016-08-01

    During embryogenesis, organisms undergo considerable cellular remodelling requiring the combined action of thousands of proteins. In case of the well-studied model Drosophila melanogaster, transcriptomic studies, most notably from the modENCODE project, have described in detail changes in gene expression at the mRNA level across development. Although such data are clearly very useful to understand how the genome is regulated during embryogenesis, it is important to understand how changes in gene expression are reflected at the level of the proteome. In this study, we describe a combination of two quantitative label-free approaches, SWATH and data-dependent acquisition, to monitor changes in protein expression across a timecourse of D. melanogaster embryonic development. We demonstrate that both approaches provide robust and reproducible methods for the analysis of proteome changes. In a preliminary analysis of Drosophila embryogenesis, we identified several pathways, including the heat-shock response, nuclear protein import and energy production that are regulated during embryo development. In some cases changes in protein expression mirrored transcript levels across development, whereas other proteins showed signatures of post-transcriptional regulation. Taken together, our pilot study provides a solid platform for a more detailed exploration of the embryonic proteome. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. A graphene-based label-free voltammetric immunosensor for sensitive detection of the egg allergen ovalbumin.

    Science.gov (United States)

    Eissa, Shimaa; L'Hocine, Lamia; Siaj, Mohamed; Zourob, Mohammed

    2013-08-07

    A graphene-based label-free voltammetric immunosensor for the sensitive detection of the egg white allergen ovalbumin has been developed. Graphene-modified screen printed carbon electrodes have been covalently functionalized using electrochemical reduction of in situ generated aryl diazonium salt forming a carboxyphenyl film on the graphene surface. The blocking property of the carboxyphenyl film grafted on to the graphene electrodes using different cyclic voltammetry cycles has been characterized using differential pulse voltammetry in [Fe(CN)6](3-/4-) solution. Then, the terminal carboxylic groups on the graphene surface were activated using EDC/NHS and used to immobilize the ovalbumin antibody and construct the immunosensor. The fabrication steps of the immunosensor have also been characterized using differential pulse voltammetry. The decrease in the [Fe(CN)6](3-/4-) reduction peak current after the immunochemical reaction with ovalbumin has been used for the ovalbumin detection. The developed immunosensor has been used for ovalbumin detection in the concentration range of 1 pg mL(-1) to 0.5 μg mL(-1) with a detection limit of 0.83 pg mL(-1) in PBS buffer. The food matrix effect studied with ovalbumin spiked cake extract showed a good percentage of recovery, indicating the possible applicability of the developed immunosensor in real food samples.

  7. Single-walled carbon nanotubes based chemiresistive genosensor for label-free detection of human rheumatic heart disease

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Swati; Kumar, Ashok, E-mail: rajesh-csir@yahoo.com, E-mail: ashokigib@rediffmail.com [CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi 110007 (India); Academy of Scientific and Innovative Research (AcSIR), New Delhi (India); Khare, Shashi [National Centre for Disease Control, Sham Nath Marg, Delhi 110054 (India); Mulchandani, Ashok [Department of Chemical and Environmental Engineering, University of California, Riverside, California 92521 (United States); Rajesh, E-mail: rajesh-csir@yahoo.com, E-mail: ashokigib@rediffmail.com [CSIR-National Physical Laboratory, Dr. K. S. Krishnan Road, New Delhi 110012 (India)

    2014-11-24

    A specific and ultrasensitive, label free single-walled carbon nanotubes (SWNTs) based chemiresistive genosensor was fabricated for the early detection of Streptococcus pyogenes infection in human causing rheumatic heart disease. The mga gene of S. pyogenes specific 24 mer ssDNA probe was covalently immobilized on SWNT through a molecular bilinker, 1-pyrenemethylamine, using carbodiimide coupling reaction. The sensor was characterized by the current-voltage (I-V) characteristic curve and scanning electron microscopy. The sensing performance of the sensor was studied with respect to changes in conductance in SWNT channel based on hybridization of the target S. pyogenes single stranded genomic DNA (ssG-DNA) to its complementary 24 mer ssDNA probe. The sensor shows negligible response to non-complementary Staphylococcus aureus ssG-DNA, confirming the specificity of the sensor only with S. pyogenes. The genosensor exhibited a linear response to S. pyogenes G-DNA from 1 to1000 ng ml{sup −1} with a limit of detection of 0.16 ng ml{sup −1}.

  8. A carbon nanotube screen-printed electrode for label-free detection of the human cardiac troponin T.

    Science.gov (United States)

    Silva, Bárbara V M; Cavalcanti, Igor T; Silva, Mízia M S; Dutra, Rosa F

    2013-12-15

    Label-free immunosensor based on amine-functionalized carbon nanotubes screen-printed electrode is described for detection of the cardiac troponin T, an important marker of acute myocardial infarction. The disposable sensor was fabricated by tightly squeezing an adhesive carbon ink containing carbon nanotubes onto a polyethylene terephthalate substrate forming a thin film. The use of carbon nanotubes increased the reproducibility and stability of the sensor, and the amine groups permitted nonrandom immobilization of antibodies against cardiac troponin T. Amperometric responses were obtained by differential pulse voltammetry in presence of a ferrocyanide/ferricyanide redox probe after troponin T incubation. The calibration curve indicated a linear response of troponin T between 0.0025 ng mL(-1) and 0.5 ng mL(-1), with a good correlation coefficient (r=0.995; p<0.0001, n=7). The limit of detection (0.0035 ng mL(-1) cardiac troponin T) was lower than any previously described by immunosensors and was comparable with conventional analytical methods. The high reproducibility and clinical range obtained using this immunosensor support its utility as a potential tool for point-of-care acute myocardial infarction diagnostic testing. © 2013 Elsevier B.V. All rights reserved.

  9. In-depth evaluation of software tools for data-independent acquisition based label-free quantification.

    Science.gov (United States)

    Kuharev, Jörg; Navarro, Pedro; Distler, Ute; Jahn, Olaf; Tenzer, Stefan

    2015-09-01

    Label-free quantification (LFQ) based on data-independent acquisition workflows currently experiences increasing popularity. Several software tools have been recently published or are commercially available. The present study focuses on the evaluation of three different software packages (Progenesis, synapter, and ISOQuant) supporting ion mobility enhanced data-independent acquisition data. In order to benchmark the LFQ performance of the different tools, we generated two hybrid proteome samples of defined quantitative composition containing tryptically digested proteomes of three different species (mouse, yeast, Escherichia coli). This model dataset simulates complex biological samples containing large numbers of both unregulated (background) proteins as well as up- and downregulated proteins with exactly known ratios between samples. We determined the number and dynamic range of quantifiable proteins and analyzed the influence of applied algorithms (retention time alignment, clustering, normalization, etc.) on quantification results. Analysis of technical reproducibility revealed median coefficients of variation of reported protein abundances below 5% for MS(E) data for Progenesis and ISOQuant. Regarding accuracy of LFQ, evaluation with synapter and ISOQuant yielded superior results compared to Progenesis. In addition, we discuss reporting formats and user friendliness of the software packages. The data generated in this study have been deposited to the ProteomeXchange Consortium with identifier PXD001240 (http://proteomecentral.proteomexchange.org/dataset/PXD001240). © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Label-free serum ribonucleic acid analysis for colorectal cancer detection by surface-enhanced Raman spectroscopy and multivariate analysis

    Science.gov (United States)

    Chen, Yanping; Chen, Gang; Feng, Shangyuan; Pan, Jianji; Zheng, Xiongwei; Su, Ying; Chen, Yan; Huang, Zufang; Lin, Xiaoqian; Lan, Fenghua; Chen, Rong; Zeng, Haishan

    2012-06-01

    Studies with circulating ribonucleic acid (RNA) not only provide new targets for cancer detection, but also open up the possibility of noninvasive gene expression profiling for cancer. In this paper, we developed a surface-enhanced Raman scattering (SERS), platform for detection and differentiation of serum RNAs of colorectal cancer. A novel three-dimensional (3-D), Ag nanofilm formed by dry MgSO4 aggregated silver nanoparticles, Ag NP, as the SERS-active substrate was presented to effectively enhance the RNA Raman signals. SERS measurements were performed on two groups of serum RNA samples. One group from patients, n=55 with pathologically diagnosed colorectal cancer and the other group from healthy controls, n=45. Tentative assignments of the Raman bands in the normalized SERS spectra demonstrated that there are differential expressions of cancer-related RNAs between the two groups. Linear discriminate analysis, based on principal component analysis, generated features can differentiate the colorectal cancer SERS spectra from normal SERS spectra with sensitivity of 89.1 percent and specificity of 95.6 percent. This exploratory study demonstrated great potential for developing serum RNA SERS analysis into a useful clinical tool for label-free, noninvasive screening and detection of colorectal cancers.

  11. Detection of ESAT-6 by a label free miniature immuno-electrochemical biosensor as a diagnostic tool for tuberculosis.

    Science.gov (United States)

    Diouani, Mohamed Fethi; Ouerghi, Oussama; Refai, Amira; Belgacem, Kamel; Tlili, Chaker; Laouini, Dhafer; Essafi, Makram

    2017-05-01

    Tuberculosis is a worldwide disease considered as a major health problem with high morbidity and mortality rates. Poor detection of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis remains a major obstacle to the global control of this disease. Here we report the development of a new test based on the detection of the major virulent factor of Mtb, namely the early secreted antigenic target 6-kDa protein or ESAT-6. A label free electrochemical immunosensor using an anti-ESAT-6 monoclonal antibody as a bio-receptor is described herein. Anti-ESAT-6 antibodies were first covalently immobilized on the surface of a gold screen-printed electrode functionalized via a self-assembled thiol monolayer. Interaction between the bio-receptor and ESAT-6 antigen was evaluated by square wave voltammetry method using [F e (CN) 6 ] 3-/4- as redox probe. The detection limit of ESAT-6 antigen was 7ng/ml. The immunosensor has also been able to detect native ESAT-6 antigen secreted in cell culture filtrates of three pathogenic strains of Mtb (CDC1551, H37RV and H8N8). Overall, this work describes an immune-electrochemical biosensor, based on ESAT-6 antigen detection, as a useful diagnostic tool for tuberculosis. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Single-Cell Optical Distortion Correction and Label-Free 3D Cell Shape Reconstruction on Lattices of Nanostructures.

    Science.gov (United States)

    Stephan, Jürgen; Keber, Felix; Stierle, Valentin; Rädler, Joachim O; Paulitschke, Philipp

    2017-12-13

    Imaging techniques can be compromised by aberrations. Especially when imaging through biological specimens, sample-induced distortions can limit localization accuracy. In particular, this phenomenon affects localization microscopy, traction force measurements, and single-particle tracking, which offer high-resolution insights into biological tissue. Here we present a method for quantifying and correcting the optical distortions induced by single, adherent, living cells. The technique uses periodically patterned gold nanostructures as a reference framework to quantify optically induced displacements with micrometer-scale sampling density and an accuracy of a few nanometers. The 3D cell shape and a simplified geometrical optics approach are then utilized to remap the microscope image. Our experiments reveal displacements of up to several hundred nanometers, and in corrected images these distortions are reduced by a factor of 3. Conversely, the relationship between cell shape and distortion provides a novel method of 3D cell shape reconstruction from a single image, enabling label-free 3D cell analysis.

  13. Surface stress sensor based on MEMS Fabry-Perot interferometer with high wavelength selectivity for label-free biosensing

    Science.gov (United States)

    Takahashi, Toshiaki; Hizawa, Takeshi; Misawa, Nobuo; Taki, Miki; Sawada, Kazuaki; Takahashi, Kazuhiro

    2018-05-01

    We have developed a surface stress sensor based on a microelectromechanical Fabry-Perot interferometer with high wavelength selectivity by using Au half-mirrors, for highly sensitive label-free biosensing. When the target molecule is adsorbed by the antigen-antibody reaction onto a movable membrane with a thin Au film, which acts as an upper mirror of the optical interferometer, the amount of deflection of the movable membrane deflected by the change in surface stress can be detected with high sensitivity. To improve the signal at the small membrane deflection region of this biosensor resulting in detection of low concentration molecules, by integrating 50 nm-thick Au half-mirrors, the wavelength selectivity of the optical interferometer has been successfully improved 6.6 times. Furthermore, the peak shift in the reflection spectrum due to the adsorption of bovine serum albumin (BSA) antigen with a concentration of 10 ng ml-l by the antigen-antibody reaction was spectroscopically measured on the fabricated optical interferometer, and the deflection amount of the movable membrane after 10 min treatment was 2.4 times larger than that of nonspecific adsorption with the avidin molecules. This result indicated that the proposed sensor can be used for selective detection of low-concentration target antigen molecules.

  14. Quantitative evaluation of alternatively spliced mRNA isoforms by label-free real-time plasmonic sensing.

    Science.gov (United States)

    Huertas, César S; Carrascosa, L G; Bonnal, S; Valcárcel, J; Lechuga, L M

    2016-04-15

    Alternative splicing of mRNA precursors enables cells to generate different protein outputs from the same gene depending on their developmental or homeostatic status. Its deregulation is strongly linked to disease onset and progression. Current methodologies for monitoring alternative splicing demand elaborate procedures and often present difficulties in discerning between closely related isoforms, e.g. due to cross-hybridization during their detection. Herein, we report a general methodology using a Surface Plasmon Resonance (SPR) biosensor for label-free monitoring of alternative splicing events in real-time, without any cDNA synthesis or PCR amplification requirements. We applied this methodology to RNA isolated from HeLa cells for the quantification of alternatively spliced isoforms of the Fas gene, involved in cancer progression through regulation of programmed cell death. We demonstrate that our methodology is isoform-specific, with virtually no cross-hybridization, achieving limits of detection (LODs) in the picoMolar (pM) range. Similar results were obtained for the detection of the BCL-X gene mRNA isoforms. The results were independently validated by RT-qPCR, with excellent concordance in the determination of isoform ratios. The simplicity and robustness of this biosensor technology can greatly facilitate the exploration of alternative splicing biomarkers in disease diagnosis and therapy. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element

    Directory of Open Access Journals (Sweden)

    Özlem Ertekin

    2016-08-01

    Full Text Available This study introduces the use of an IgA isotype aflatoxin (AF specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl carbodiimide/N-hydroxy succinimide (EDC/NHS method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1.

  16. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element.

    Science.gov (United States)

    Ertekin, Özlem; Öztürk, Selma; Öztürk, Zafer Ziya

    2016-08-11

    This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1.

  17. Label-free electrochemical immunoassay for neuron specific enolase based on 3D macroporous reduced graphene oxide/polyaniline film.

    Science.gov (United States)

    Zhang, Qi; Li, Xiaoyan; Qian, Chunhua; Dou, Li; Cui, Feng; Chen, Xiaojun

    2018-01-01

    The content of neuron specific enolase (NSE) in serum is considered to be an essential indicator of small cell lung cancer (SCLC). Here, a novel label-free electrochemical immunoassay for the detection of NSE based on the three dimensionally macroporous reduced graphene oxide/polyaniline (3DM rGO/PANI) film has been proposed. The 3DM rGO/PANI film was constructed by electrochemical co-deposition of GO and aniline into the interspaces of a sacrificial silica opal template modified Au slice. During the co-deposition, GO was successfully reduced by aniline and PANI could be deposited on the surfaces of rGO sheets. The ratio of rGO and PANI in the composite was also optimized to achieve the maximum electrochemical performance. The 3DM rGO/PANI composite provided larger specific surface area for the antibody immobilization, exhibited enhanced conductivity for electron transfer, and more important was that PANI acted as the electroactive probe for indicating the NSE concentration. Under the optimal conditions, a linear current response of PANI to NSE concentration was obtained over 0.5 pg mL -1 -10.0 ng mL -1 with a detection limit of 0.1 pg mL -1 . Moreover, the immunosensor showed excellent selectivity, good stability, satisfactory reproducibility and regeneration, and was employed to detect NSE in clinical serum specimens. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. A silicon-based electrochemical sensor for highly sensitive, specific, label-free and real-time DNA detection

    International Nuclear Information System (INIS)

    Guo, Yuanyuan; Su, Shao; Wei, Xinpan; Zhong, Yiling; Su, Yuanyuan; He, Yao; Huang, Qing; Fan, Chunhai

    2013-01-01

    We herein present a new kind of silicon-based electrochemical sensor using a gold nanoparticles-decorated silicon wafer (AuNPs@Si) as a high-performance electrode, which is facilely prepared via in situ AuNPs growth on a silicon wafer. Particularly significantly, the resultant electrochemical sensor is efficacious for label-free DNA detection with high sensitivity due to the unique merits of the prepared silicon-based electrode. Typically, DNA at remarkably low concentrations (1–10 fM) could be readily detected without requiring additional signal-amplification procedures, which is better than or comparable to the lowest DNA concentration ever detected via well-studied signal-amplification-assisted electrochemical sensors. Moreover, the silicon-based sensor features high specificity, allowing unambiguous discrimination of single-based mismatches. We further show that real-time DNA assembly is readily monitored via recording the intensity changes of current signals due to the robust thermal stability of the silicon-based electrode. The unprecedented advantages of the silicon-based electrochemical sensor would offer new opportunities for myriad sensing applications. (paper)

  19. Label-free detection of kanamycin based on a G-quadruplex DNA aptamer-based fluorescent intercalator displacement assay

    Science.gov (United States)

    Xing, Yun-Peng; Liu, Chun; Zhou, Xiao-Hong; Shi, Han-Chang

    2015-01-01

    This work was the first to report that the kanamycin-binding DNA aptamer (5'-TGG GGG TTG AGG CTA AGC CGA-3') can form stable parallel G-quadruplex DNA (G4-DNA) structures by themselves and that this phenomenon can be verified by nondenaturing polyacrylamide gel electrophoresis and circular dichroism spectroscopy. Based on these findings, we developed a novel label-free strategy for kanamycin detection based on the G4-DNA aptamer-based fluorescent intercalator displacement assay with thiazole orange (TO) as the fluorescence probe. In the proposed strategy, TO became strongly fluorescent upon binding to kanamycin-binding G4-DNA. However, the addition of kanamycin caused the displacement of TO from the G4-DNA-TO conjugate, thereby resulting in decreased fluorescent signal, which was inversely related to the kanamycin concentration. The detection limit of the proposed assay decreased to 59 nM with a linear working range of 0.1 μM to 20 μM for kanamycin. The cross-reactivity against six other antibiotics was negligible compared with the response to kanamycin. A satisfactory recovery of kanamycin in milk samples ranged from 80.1% to 98.0%, confirming the potential of this bioassay in the measurement of kanamycin in various applications. Our results also served as a good reference for developing similar fluorescent G4-DNA-based bioassays in the future.

  20. Foreign direct investment (FDI) in energy/electricity projects in the economies in transition

    International Nuclear Information System (INIS)

    Brendow, K.

    1995-01-01

    Registered foreign direct investments (FDI) in energy/ electricity projects in central and eastern Europe and the CIS are weak and disappointing compared with expectations and business opportunities. In addition, energy FDI is highly concentrated in oil and gas exploration/extraction. FDI opportunities in electricity generation, transmission and distribution while being explored, have not been implemented, mostly for legislation restriction reasons towards foreign participation. The major issue appears to be the extent of privatization of this politically and socially sensitive industry. Progress are made in Hungary, Poland and the Czech Republic. A significant breakthrough depends on political stability, economic recovery and progress in privatization. 2 figs

  1. The directive on ''internal market of electric power'': a satisfactory balance

    International Nuclear Information System (INIS)

    1996-01-01

    The european directive on the ''internal market for electric power'' has allowed for the first time the implementation of harmonized regulations among european partners concerning the operation of electric utilities and systems in Europe, while taking into consideration the subsidiarity principles to which France is very much attached and finding an equilibrium between the status-quo based on national or regional monopolies and a global deregulation inspired by ultra-liberal views coming from Britain or the US. The main consequences on France's energy policy and organization are discussed

  2. Influence of Direct Current Electric Field on Corrosion Behavior of Tin Under a Thin Electrolyte Layer

    Science.gov (United States)

    Huang, H. L.; Bu, F. R.; Tian, J.; Liu, D.

    2017-12-01

    The influence of a direct current electric field (DCEF) on corrosion behavior of tin under a thin electrolyte layer was investigated based on an array electrode technology by polarization, electrochemical impedance spectroscopy and surface analysis. The experimental results indicate that the corrosion rate of tin near the positive plate of DCEF increases with increased electric field intensity, which could be attributed to the acceleration of the migration of ions, the removal of corrosion products under DCEF and the damage of tin surface oxide film. Furthermore, tin at different positions in a DCEF exhibits different corrosion behavior, which could be ascribed to the difference of the local corrosion environment caused by the DCEF.

  3. Effects of the Electric Field on the Direct Loss Asymmetries on TJ-II

    International Nuclear Information System (INIS)

    Guasp, J.

    1997-01-01

    The effects of the radial electric fields upon the direct loss asymmetries have been studied for low energy (0.1-1keV)ions. The analysis shows the permanence of the predominant downwards asymmetry that is only slightly modified in the poloidal rotation direction for moderate electric fields. In TJ-II this poloidal rotation is directed towards decreasing poloidal angles for positive (outward) electric fields and the ions drift along the same direction. Nevertheless for very strong fields the lost particles concentrate in the extreme plasma periphery (at the same time that the rate loss decreases strongly) and have no time, any more, to feel the poloidal rotation before escaping. The gaps between the TF coils receive the most of the losses. On the Vacuum Vessel the losses have a very strong concentration along the Hard Core, in particular on the PLT-2 plane plate (the one that is placed down at Phi=0 degree centigree) and a moderate preference for the 2''nd toroidal octant of each period. The poloidal rotation is much less visible than on the plasma border. The resulting power loads are small. (Author)

  4. High electrical conductivity in out of plane direction of electrodeposited Bi2Te3 films

    Directory of Open Access Journals (Sweden)

    Miguel Muñoz Rojo

    2015-08-01

    Full Text Available The out of plane electrical conductivity of highly anisotropic Bi2Te3 films grown via electro-deposition process was determined using four probe current-voltage measurements performed on 4.6 - 7.2 μm thickness Bi2Te3 mesa structures with 80 - 120 μm diameters sandwiched between metallic film electrodes. A three-dimensional finite element model was used to predict the electric field distribution in the measured structures and take into account the non-uniform distribution of the current in the electrodes in the vicinity of the probes. The finite-element modeling shows that significant errors could arise in the measured film electrical conductivity if simpler one-dimensional models are employed. A high electrical conductivity of (3.2 ± 0.4 ⋅ 105 S/m is reported along the out of plane direction for Bi2Te3 films highly oriented in the [1 1 0] direction.

  5. Direct Measurement of Bipolar Cell Responses to Electrical Stimulation in Wholemount Mouse Retina.

    Science.gov (United States)

    Walston, Steven T; Chow, Robert H; Weiland, James D

    2018-03-07

    This in vitro investigation examines the response of retinal bipolar cells to extracellular electrical stimulation. Approach: In vitro investigations characterizing the response of retinal neurons to electrical stimulation have primarily focused on retinal ganglion cells because they are the output neurons of the retina and their superficial position in the retina makes them readily accessible to in vitro recording techniques. Thus, the majority of information regarding the response of inner retinal neurons has been inferred from ganglion cell activity. Here we use patch clamp electrophysiology to directly record electrically-evoked activity in bipolar cells within the inner retina of normal Tg(Gng13-EGFP)GI206Gsat and degenerate rd10 Tg(Gng13-EGFP)GI206Gsat mice using a wholemount preparation. Main Results: Bipolar cells respond to electrical stimulation with time-locked depolarizing voltage transients. The latency of the response declines with increases in stimulation amplitude. A desensitizing response is observed during repeated stimulation with 25-ms biphasic current pulses delivered at pulse rates greater than 6 pps. A burst of long-latency (200-1000 ms) inhibitory postsynaptic potentials are evoked by the stimulus and the burst exhibits evidence of a lower and upper stimulation threshold. Significance: These results provide insights into the various types of bipolar cell activity elicited by electrical stimulation and may be useful for future retinal prosthesis stimulation protocols. . © 2018 IOP Publishing Ltd.

  6. Direct nanoscale imaging of evolving electric field domains in quantum structures.

    Science.gov (United States)

    Dhar, Rudra Sankar; Razavipour, Seyed Ghasem; Dupont, Emmanuel; Xu, Chao; Laframboise, Sylvain; Wasilewski, Zbig; Hu, Qing; Ban, Dayan

    2014-11-28

    The external performance of quantum optoelectronic devices is governed by the spatial profiles of electrons and potentials within the active regions of these devices. For example, in quantum cascade lasers (QCLs), the electric field domain (EFD) hypothesis posits that the potential distribution might be simultaneously spatially nonuniform and temporally unstable. Unfortunately, there exists no prior means of probing the inner potential profile directly. Here we report the nanoscale measured electric potential distribution inside operating QCLs by using scanning voltage microscopy at a cryogenic temperature. We prove that, per the EFD hypothesis, the multi-quantum-well active region is indeed divided into multiple sections having distinctly different electric fields. The electric field across these serially-stacked quantum cascade modules does not continuously increase in proportion to gradual increases in the applied device bias, but rather hops between discrete values that are related to tunneling resonances. We also report the evolution of EFDs, finding that an incremental change in device bias leads to a hopping-style shift in the EFD boundary--the higher electric field domain expands at least one module each step at the expense of the lower field domain within the active region.

  7. Direct current electric field assembly of colloidal crystals displaying reversible structural color.

    Science.gov (United States)

    Shah, Aayush A; Ganesan, Mahesh; Jocz, Jennifer; Solomon, Michael J

    2014-08-26

    We report the application of low-voltage direct current (dc) electric fields to self-assemble close-packed colloidal crystals in nonaqueous solvents from colloidal spheres that vary in size from as large as 1.2 μm to as small as 0.1 μm. The assemblies are created rapidly (∼2 min) from an initially low volume fraction colloidal particle suspension using a simple capacitor-like electric field device that applies a steady dc electric voltage. Confocal microscopy is used to observe the ordering that is produced by the assembly method. This spatial evidence for ordering is consistent with the 6-fold diffraction patterns identified by light scattering. Red, green, and blue structural color is observed for the ordered assemblies of colloids with diameters of 0.50, 0.40, and 0.29 μm, respectively, consistent with spectroscopic measurements of reflectance. The diffraction and spectrophotometry results were found to be consistent with the theoretical Bragg's scattering expected for closed-packed crystals. By switching the dc electric field from on to off, we demonstrate reversibility of the structural color response on times scales ∼60 s. The dc electric field assembly method therefore represents a simple method to produce reversible structural color in colloidal soft matter.

  8. Direct electrical stimulation of human cortex evokes high gamma activity that predicts conscious somatosensory perception

    Science.gov (United States)

    Muller, Leah; Rolston, John D.; Fox, Neal P.; Knowlton, Robert; Rao, Vikram R.; Chang, Edward F.

    2018-04-01

    Objective. Direct electrical stimulation (DES) is a clinical gold standard for human brain mapping and readily evokes conscious percepts, yet the neurophysiological changes underlying these percepts are not well understood. Approach. To determine the neural correlates of DES, we stimulated the somatosensory cortex of ten human participants at frequency-amplitude combinations that both elicited and failed to elicit conscious percepts, meanwhile recording neural activity directly surrounding the stimulation site. We then compared the neural activity of perceived trials to that of non-perceived trials. Main results. We found that stimulation evokes distributed high gamma activity, which correlates with conscious perception better than stimulation parameters themselves. Significance. Our findings suggest that high gamma activity is a reliable biomarker for perception evoked by both natural and electrical stimuli.

  9. Measurement of the transverse electric field profile of light by a self-referencing method with direct phase determination.

    Science.gov (United States)

    Bamber, C; Sutherland, B; Patel, A; Stewart, C; Lundeen, J S

    2012-01-30

    We present a method for measuring the transverse electric field profile of a beam of light which allows for direct phase retrieval. The measured values correspond, within a normalization constant, to the real and imaginary parts of the electric field in a plane normal to the direction of propagation. This technique represents a self-referencing method for probing the wavefront characteristics of light.

  10. 76 FR 292 - Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines

    Science.gov (United States)

    2011-01-04

    ...-24145; Directorate Identifier 2006-NE-06-AD] RIN 2120-AA64 Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines AGENCY: Federal Aviation Administration (FAA), DOT... airworthiness directive (AD) that applies to General Electric Company (GE) CF6-45 and CF6-50 series turbofan...

  11. 75 FR 12661 - Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines

    Science.gov (United States)

    2010-03-17

    ... Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines AGENCY: Federal... a new airworthiness directive (AD) for General Electric Company (GE) CF6-45 and CF6-50 series turbofan engines with certain low-pressure turbine (LPT) stage 3 disks installed. This AD requires...

  12. 75 FR 32649 - Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines

    Science.gov (United States)

    2010-06-09

    ... Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines AGENCY: Federal Aviation... airworthiness directive (AD) for General Electric Company (GE) CF6-45 and CF6-50 series turbofan engines with... issuing this AD to prevent critical life-limited rotating engine part failure, which could result in an...

  13. 76 FR 14796 - Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines

    Science.gov (United States)

    2011-03-18

    ... Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines AGENCY: Federal... airworthiness directive (AD) for General Electric Company (GE) CF6-45 and CF6-50 series turbofan engines. That... the LFCEN assembly from separating from the engine, causing damage to the engine, and damage to the...

  14. 76 FR 6323 - Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines

    Science.gov (United States)

    2011-02-04

    ... Airworthiness Directives; General Electric Company CF6-45 and CF6-50 Series Turbofan Engines AGENCY: Federal... an existing airworthiness directive (AD) for General Electric Company (GE) CF6-45 and CF6-50 series turbofan engines with certain low-pressure turbine (LPT) rotor stage 3 disks installed. That AD currently...

  15. Starting assistance device for directly-driven electric vehicle with variable-idle speed control

    OpenAIRE

    Xiong, Ping; Gu, Chenglin

    2017-01-01

    By introducing a novel permanent magnetic bi-stable electromagnetic clutch unit (PMBECU) in the driveline of directly-driven electric vehicle, a flexible connection between motor and hub is realized with characteristic of separated starting process of drive motor and wheel load. Firstly, the working principle and power supply scheme for the PMBECU is discussed. Then the assembly for the wheel-drive system with starting assistance device by the clutch is investigated. Considering the good dyna...

  16. Pyrrolidinyl PNA polypyrrole/silver nanofoam electrode as a novel label-free electrochemical miRNA-21 biosensor.

    Science.gov (United States)

    Kangkamano, Tawatchai; Numnuam, Apon; Limbut, Warakorn; Kanatharana, Proespichaya; Vilaivan, Tirayut; Thavarungkul, Panote

    2018-04-15

    A label-free electrochemical miRNA biosensor was developed based on a pyrrolidinyl peptide nucleic acid (acpcPNA)/polypyrrole (PPy)/silver nanofoam (AgNF) modified electrode. The AgNF was electrodeposited as redox indicator on a gold electrode, which was then functionalized with an electropolymerized layer of PPy, a conducting polymer, to immobilize the PNA probes. The fabrication process was investigated by electrochemical impedance spectroscopy. The biosensor was used to detect miRNA-21, a biomarker abnormally expressed in most cancers. The signal was monitored by the change in current of the AgNF redox reaction before and after hybridization using cyclic voltammetry. Two PNA probe lengths were investigated and the longer probe exhibited a better performance. Nucleotide overhangs on the electrode side affected the signal more than overhangs on the solution side due to the greater insulation of the sensing surface. Under optimal conditions, the electrochemical signal was proportional to miRNA-21 concentrations between 0.20fM and 1.0nM, with a very low detection limit of 0.20fM. The biosensor showed a high specificity which could discriminate between complementary, single-, doubled-base mismatched, and non-complementary targets. Three out of the seven tested plasma samples provided detectable concentrations (63 ± 4, 111 ± 4 and 164 ± 7fM). The sensor also showed good recoveries (81-119%). The results indicated the possibilities of this biosensor for analysis without RNA extraction and/or amplification, making the sensor potentially useful for both the prognosis and diagnosis of cancer in clinical application. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Coreactant-free and Label-free Eletrochemiluminescence Immunosensor for Copeptin Based on Luminescent Immuno-Gold Nanoassemblys.

    Science.gov (United States)

    Han, Zhili; Shu, Jiangnan; Jiang, Qiaoshi; Cui, Hua

    2018-04-25

    In this work, the eletrochemiluminescence (ECL) behavior of Cu 2+ /cysteine complexes and N-(aminobutyl)-N-(ethylisoluminol) (ABEI) functionalized gold nanoparticles combined with chitosan (Cu 2+ -Cys-ABEI-GNPs-CS) were studied by cyclic voltammetry and a double-step potential, which exhibited excellent ECL properties without any coreactant. It was found that the ECL intensity of Cu 2+ -Cys-ABEI-GNPs-CS could increase at least one order of magnitude compared with that of Cu 2+ -Cys-ABEI-GNPs. Furthermore, a coreactant-free and label-free ECL immunosensor has been established for the determination of early acute myocardial infarction biomarker copeptin based on luminescent immuno-gold nanoassemblys consisting of Cu 2+ -Cys-ABEI-GNPs-CS and immuno-gold nanoparticles prepared by connecting copeptin antibody with trisodium citrate stabilized gold nanoparticles. In the presence of copeptin, an obvious decrease in ECL intensity was observed due to the formation of antibody-antigen immunocomplex, which could be used for the determination of copeptin in the range of 2.0×10 -14 -1.0×10 -11 mol/L with a detection limit of 5.18×10 -15 mol/L. The detection limit of the ECL immunosensor is at least two orders of magnitude lower than that of sandwich immunoassays based on labeling technology. And the ECL immunosensor does not need any coreactant, and avoids complicated labeling and purification procedure. It is ultrasensitive, simple, specific and low-cost. This work reveals that the proposed luminescent immuno-gold nanoassemblys are ideal nanointerfaces for the construction of immunosensors. The proposed strategy may be used for the determination of other antigens if corresponding antibodies are available.

  18. Label-free quantitative proteomic analysis reveals strong involvement of complement alternative and terminal pathways in human glomerular sclerotic lesions.

    Science.gov (United States)

    Zhang, Ying; Xu, Bo; Kinoshita, Naohiko; Yoshida, Yutaka; Tasaki, Masayuki; Fujinaka, Hidehiko; Magdeldin, Sameh; Yaoita, Eishin; Yamamoto, Tadashi

    2015-06-18

    Since glomerular sclerosis frequently accompanies various glomerular diseases at the end stages, it is challenging to differentiate ubiquitous biological processes underlying this pathology from those critically involved in specific diseases. Furthermore, in-depth proteomic profile of human glomerular sclerosis remains limited. In this study, human glomeruli with intermediate (i-GS) and advanced (GS) sclerotic lesions, which were excluded from specific renal diseases and assumed to be aging-related, were laser captured from macroscopically normal cortex distant from urological carcinoma, and subjected to label-free quantitative proteomic analysis. We explicate an evident increase of membrane attack complex in i-GS and GS with an up-going tendency, which is accompanied by increasing of inhibitory regulators of alternative and terminal pathways. GO annotation and IPA pathway analysis agree to these results. Proteomic findings are validated by immunohistochemical studies which indicate that alternative and terminal pathways are positively involved in the glomerular sclerosis seen in distinct renal diseases. Furthermore, proteomic analysis also demonstrates remarkable increases of complement factor B in GS and TGF-ß1 in both GS and i-GS. Identification of complement factor B implicates that on-site activation of alternative pathway may occur in injured glomeruli and stepwise increase of TGF-ß1 suggests its contribution to the progression of glomerulosclerosis. This study provides in-depth quantitative proteomic profiles of human glomeruli with intermediate and advanced sclerotic lesions. It reveals that the over-expression of alternative and terminal pathway components is significantly involved in human glomerulosclerosis seen in distinct renal diseases. Proteomic identification of the increased TGF-ß1 provides supporting evidence for the role of podocyte apoptosis leading to human glomerulosclerosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. A Raman Flow Cytometer: An Innovative Microfluidic Approach for Continuous Label-Free Analysis of Cells via Raman Spectroscopy

    KAUST Repository

    De Grazia, Antonio

    2015-05-05

    In this work a Raman flow cytometer is presented. It is a whole new microfluidic device that takes advantage of basic principles of Raman spectroscopy and fluorescent flow cytometry mixed together in a system of particularly shaped channels. These are indeed composed by specific shape and sizes – thanks to which cells can flow one-by-one – and a trap by means of which cells are trapped in order to perform Raman analysis on single ones in a constant and passive way. In this sense the microfluidic device promotes a fast method to look for single cells in a whole multicellular sample. It is a label-free analysis and this means that, on the contrary of what happens with fluorescent flow cytometry, the sample does not need to undergo any particular time-consuming pretreatment before being analyzed. Moreover it gives a complete information about the biochemical content of the sample thanks to the involvement of Raman spectroscopy as method of analysis. Many thought about a device like this, but eventually it is the first one being designed, fabricated and tested. The materials involved in the production of the Raman flow cytometer are chosen wisely. In particular the chip – the most important component of the device – is multilayered, being composed by a slide of calcium fluoride (which gives a negligible signal in Raman analyses), a photosensitive resist containing a pattern with channels and another slide of calcium fluoride in order for the channels to be sealed on both sides. The chip is, in turn, connected to gaskets and external frames. Several fabrication processes are followed to ultimately get the complete Raman flow cytometer and experiments on red blood cells demonstrate its validity in this field.

  20. Cost-effective and label-free holographic biosensor for detection of herpes simplex virus (Conference Presentation)

    Science.gov (United States)

    Ray, Aniruddha; Ho, Ha; Daloglu, Mustafa; Torres, Avee; McLeod, Euan; Ozcan, Aydogan

    2017-03-01

    Herpes is one of the most widespread sexually transmitted viral diseases. Timely detection of Herpes Simplex Virus (HSV) can help prevent the rampant spreading of the virus. Current detection techniques such as viral culture, immuno-assays or Polymerase-Chain-Reaction, are time extensive and require expert handling. Here we present a field-portable, easy-to-use, and cost-effective biosensor for the detection of HSV based on holographic imaging. The virus is first captured from a target solution onto specifically developed substrates, prepared by coating glass coverslips with HSV-specific antibodies, and imaged using a lensfree holographic microscope. Several light-emitting-diodes (LEDs), coupled to multi-mode optical-fibers, are used to illuminate the sample containing the viruses. A micro-controller is used to activate the LEDs one at a time and in-line holograms are recorded using a CMOS imager placed immediately above the substrate. These sub-pixel shifted holograms are used to generate a super-resolved hologram, which is reconstructed to obtain the phase and amplitude images of the viruses. The signal of the viruses is enhanced using self-assembled PEG-based nanolenses, formed around the viral particles. Based on the phase information of the reconstructed images we can estimate the size of the viral particles, with an accuracy of +/- 11 nm, as well as quantify the viral load. The limit-of-detection of this system is estimated to be <500 viral copies per 100 μL sample volume that is imaged over 30 mm^2 field-of-view. This holographic microscopy based biosensor is label-free, cost-effective and field-portable, providing results in 2 hours, including sample preparation and imaging time.

  1. A versatile label-free and signal-on electrochemical biosensing platform based on triplex-forming oligonucleotide probe.

    Science.gov (United States)

    Wang, Xiuzhong; Jiang, Aiwen; Hou, Ting; Li, Feng

    2015-08-26

    Nucleic acid and protein assays are very important in modern life sciences, and the recently developed triplex-forming oligonucleotide probes provide a unique means for biological analysis of different kinds of analytes. Herein, we report a label-free and signal-on electrochemical sensor for the detection of specific targets, which is based on the triple-helix structure formation between the hairpin molecular beacon and the capture probe through the intermolecular DNA hybridization induced by Watson-Crick and Hoogsteen base pairings. Upon the introduction of a specific target, the triple-helical stem region is dissembled to liberate the hemin aptamer, and a G-quadruplex- hemin complex can be formed in the presence of K(+) and hemin on the electrode surface to give an electrochemical response, thus signaling the presence of the target. With the use of Human Immunodeficiency Virus type 1 (HIV-1) as a proof-of-principle analyte, we first demonstrated this approach by using a molecular beacon, which consists of a central section with the DNA sequence complementary to HIV-1, flanked by two arm segments. This newly designed protocol provides an ultrasensitive electrochemical detection of HIV-1 with a limit of detection down to 0.054 nM, and also exhibit good selectivity. Therefore, the as-proposed strategy holds a great potential for early diagnosis in gene-related diseases, and with further development, it could be used as a universal protocol for the detection of various DNA sequences and may be extended for the detection of aptamer-binding molecules. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Label-free imaging and identification of typical cells of acute myeloid leukaemia and myelodysplastic syndrome by Raman microspectroscopy.

    Science.gov (United States)

    Vanna, R; Ronchi, P; Lenferink, A T M; Tresoldi, C; Morasso, C; Mehn, D; Bedoni, M; Picciolini, S; Terstappen, L W M M; Ciceri, F; Otto, C; Gramatica, F

    2015-02-21

    In clinical practice, the diagnosis and classification of acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS) start from the manual examination of stained smears of bone marrow (BM) and peripheral blood (PB) by using an optical microscope. This step is subjective and scarcely reproducible. Therefore, the development of subjective and potentially automatable methods for the recognition of typical AML/MDS cells is necessary. Here we have used Raman spectroscopy for distinguishing myeloblasts, promyelocytes, abnormal promyelocytes and erhytroblasts, which have to be counted for a correct diagnosis and morphological classification of AML and MDS. BM samples from patients affected by four different AML subtypes, mostly characterized by the presence of the four subpopulations selected for this study, were analyzed. First, each cell was scanned by acquiring 4096 spectra, thus obtaining Raman images which demonstrate an accurate description of morphological features characteristic of each subpopulation. Raman imaging coupled with hierarchical cluster analysis permitted the automatic discrimination and localization of the nucleus, the cytoplasm, myeloperoxidase containing granules and haemoglobin. Second, the averaged Raman fingerprint of each cell was analysed by multivariate analysis (principal component analysis and linear discriminant analysis) in order to study the typical vibrational features of each subpopulation and also for the automatic recognition of cells. The leave-one-out cross validation of a Raman-based classification model demonstrated the correct classification of myeloblasts, promyelocytes (normal/abnormal) and erhytroblasts with an accuracy of 100%. Normal and abnormal promyelocytes were distinguished with 95% accuracy. The overall classification accuracy considering the four subpopulations was 98%. This proof-of-concept study shows that Raman micro-spectroscopy could be a valid approach for developing label-free, objective and automatic

  3. Shotgun label-free proteomic analysis of clubroot (Plasmodiophora brassicae resistance conferred by the gene Rcr1 in Brassica rapa

    Directory of Open Access Journals (Sweden)

    Tao Song

    2016-07-01

    Full Text Available Clubroot, caused by the plasmodiophorid pathogen Plasmodiophora brassicae, is one of the most serious diseases on Brassica crops worldwide and a major threat to canola production in western Canada. Host resistance is the key strategy for clubroot management on canola. Several clubroot resistance (CR genes have been identified, but the mechanisms associated with these CR genes are poorly understood. In the current study, a label-free shotgun proteomic approach was used to profile and compare the proteomes of B. rapa carrying and not carrying the CR gene Rcr1 upon P. brassicae infection. A total of 527 differentially accumulated proteins (DAPs were identified between the resistant and susceptible samples, and functional annotation of these DAPs indicates that the perception of P. brassicae and activation of defense responses is triggered via an unique signaling pathway distinct from common modes of recognition receptors reported with many other plant-pathogen interactions; this pathway appears to act in a calcium-independent manner through a not-well defined cascade of mitogen-activated protein kinases and may require the ubiquitin-26S proteasome related to abiotic stresses, especially the cold-stress tolerance. Both up-regulation of defense-related and down-regulation of pathogenicity-related metabolism were observed in plants carrying Rcr1, and these functions may all contribute to the clubroot resistance mediated by this CR gene. These results, combined with those of transcriptomic analysis reported earlier, improved our understanding of molecular mechanisms associated with Rcr1 and clubroot resistance at large, and identified candidate metabolites or pathways for further confirmation of specific resistance mechanisms. Deploying CR genes with different modes of action may help improve the durability of clubroot resistance.

  4. Tyramine Hydrochloride Based Label-Free System for Operating Various DNA Logic Gates and a DNA Caliper for Base Number Measurements.

    Science.gov (United States)

    Fan, Daoqing; Zhu, Xiaoqing; Dong, Shaojun; Wang, Erkang

    2017-07-05

    DNA is believed to be a promising candidate for molecular logic computation, and the fluorogenic/colorimetric substrates of G-quadruplex DNAzyme (G4zyme) are broadly used as label-free output reporters of DNA logic circuits. Herein, for the first time, tyramine-HCl (a fluorogenic substrate of G4zyme) is applied to DNA logic computation and a series of label-free DNA-input logic gates, including elementary AND, OR, and INHIBIT logic gates, as well as a two to one encoder, are constructed. Furthermore, a DNA caliper that can measure the base number of target DNA as low as three bases is also fabricated. This DNA caliper can also perform concatenated AND-AND logic computation to fulfil the requirements of sophisticated logic computing. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Label-free solution-based kinetic study of aptamer-small molecule interactions by kinetic capillary electrophoresis with UV detection revealing how kinetics control equilibrium.

    Science.gov (United States)

    Bao, Jiayin; Krylova, Svetlana M; Reinstein, Oren; Johnson, Philip E; Krylov, Sergey N

    2011-11-15

    Here we demonstrate a label-free solution-based approach for studying the kinetics of biopolymer-small molecule interactions. The approach utilizes kinetic capillary electrophoresis (KCE) separation and UV light absorption detection of the unlabeled small molecule. In this proof-of-concept work, we applied KCE-UV to study kinetics of interaction between a small molecule and a DNA aptamer. From the kinetic analysis of a series of aptamers, we found that dissociation rather than binding controls the stability of the complex. Because of its label-free features and generic nature, KCE-UV promises to become a practical tool for challenging kinetic studies of biopolymer-small molecule interactions.

  6. Directed orientation of asymmetric composite dumbbells by electric field induced assembly.

    Science.gov (United States)

    Nagao, Daisuke; Sugimoto, Maki; Okada, Ayako; Ishii, Haruyuki; Konno, Mikio; Imhof, Arnout; van Blaaderen, Alfons

    2012-04-24

    Assembly and directed orientation of anisotropic particles with an external ac electric field in a range from 1 kHz to 2 MHz were studied for asymmetric composite dumbbells incorporating a silica, titania, or titania/silica (titania:silica = 75:25 vol %) sphere. The asymmetric composite dumbbells, which were composed of a polymethylmethacrylate (PMMA)-coated sphere (core-shell part) and a polystyrene (PSt) lobe, were synthesized with a soap-free emulsion polymerization to prepare PMMA-coated inorganic spheres and another soap-free emulsion polymerization to form a polystyrene (PSt) lobe from the PMMA-coated inorganic spheres. The composite dumbbells dispersed in water were directly observed with optical microscopy. The dumbbells incorporating a silica sphere oriented parallel to an electric field in the whole frequency range and they formed a pearl chain structure at a high frequency of 2 MHz. The titania-incorporated dumbbells formed chain structures, in which they contacted their core-shell parts and oriented perpendicularly to a low-frequency (kHz) field, whereas they oriented parallel to a high-frequency (MHz) field. Since the alignment of dumbbells in the chains depends not only on the interparticle forces but also on the torque that the induced dipoles in the dumbbells experience in the electric field, the orientation of dumbbells perpendicular to the electric field was the case dominated by the interparticle force, whereas the other orientation was the case dominated by the torque. The present experiments show that the incorporation of inorganic dumbbells is an effective way to control the assembled structure and orientation with an electric field.

  7. Optically resonant magneto-electric cubic nanoantennas for ultra-directional light scattering

    Energy Technology Data Exchange (ETDEWEB)

    Sikdar, Debabrata, E-mail: debabrata.sikdar@monash.edu; Premaratne, Malin [Advanced Computing and Simulation Laboratory (A chi L), Department of Electrical and Computer Systems Engineering, Monash University, Clayton 3800, Victoria (Australia); Cheng, Wenlong [Department of Chemical Engineering, Faculty of Engineering, Monash University, Clayton 3800, Victoria (Australia); The Melbourne Centre for Nanofabrication, 151 Wellington Road, Clayton 3168, Victoria (Australia)

    2015-02-28

    Cubic dielectric nanoparticles are promising candidates for futuristic low-loss, ultra-compact, nanophotonic applications owing to their larger optical coefficients, greater packing density, and relative ease of fabrication as compared to spherical nanoparticles; besides possessing negligible heating at nanoscale in contrast to their metallic counterparts. Here, we present the first theoretical demonstration of azimuthally symmetric, ultra-directional Kerker's-type scattering of simple dielectric nanocubes in visible and near-infrared regions via simultaneous excitation and interference of optically induced electric- and magnetic-resonances up to quadrupolar modes. Unidirectional forward-scattering by individual nanocubes is observed at the first generalized-Kerker's condition for backward-scattering suppression, having equal electric- and magnetic-dipolar responses. Both directionality and magnitude of these unidirectional-scattering patterns get enhanced where matching electric- and magnetic-quadrupolar responses spectrally overlap. While preserving azimuthal-symmetry and backscattering suppression, a nanocube homodimer provides further directionality improvement for increasing interparticle gap, but with reduced main-lobe magnitude due to emergence of side-scattering lobes from diffraction-grating effect. We thoroughly investigate the influence of interparticle gap on scattering patterns and propose optimal range of gap for minimizing side-scattering lobes. Besides suppressing undesired side-lobes, significant enhancement in scattering magnitude and directionality is attained with increasing number of nanocubes forming a linear chain. Optimal directionality, i.e., the narrowest main-scattering lobe, is found at the wavelength of interfering quadrupolar resonances; whereas the largest main-lobe magnitude is observed at the wavelength satisfying the first Kerker's condition. These unique optical properties of dielectric nanocubes thus can

  8. Label-free characterization of ultra violet-radiation-induced changes in skin fibroblasts with Raman spectroscopy and quantitative phase microscopy

    OpenAIRE

    Singh, S. P.; Kang, Sungsam; Kang, Jeon Woong; So, Peter T. C.; Dasari, Ramanchandra Rao; Yaqoob, Zahid; Barman, Ishan

    2017-01-01

    Minimizing morbidities and mortalities associated with skin cancers requires sustained research with the goal of obtaining fresh insights into disease onset and progression under specific stimuli, particularly the influence of ultraviolet rays. In the present study, label-free profiling of skin fibroblasts exposed to time-bound ultra-violet radiation has been performed using quantitative phase imaging and Raman spectroscopy. Statistically significant differences in quantifiable biophysical pa...

  9. A Breast Cell Atlas: Organelle analysis of the MDA-MB-231 cell line by density-gradient fractionation using isotopic marking and label-free analysis

    Directory of Open Access Journals (Sweden)

    Marianne Sandin

    2015-09-01

    Full Text Available Protein translocation between organelles in the cell is an important process that regulates many cellular functions. However, organelles can rarely be isolated to purity so several methods have been developed to analyse the fractions obtained by density gradient centrifugation. We present an analysis of the distribution of proteins amongst organelles in the human breast cell line, MDA-MB-231 using two approaches: an isotopic labelling and a label-free approach.

  10. HoloMonitor M4: holographic imaging cytometer for real-time kinetic label-free live-cell analysis of adherent cells

    Science.gov (United States)

    Sebesta, Mikael; Egelberg, Peter J.; Langberg, Anders; Lindskov, Jens-Henrik; Alm, Kersti; Janicke, Birgit

    2016-03-01

    Live-cell imaging enables studying dynamic cellular processes that cannot be visualized in fixed-cell assays. An increasing number of scientists in academia and the pharmaceutical industry are choosing live-cell analysis over or in addition to traditional fixed-cell assays. We have developed a time-lapse label-free imaging cytometer HoloMonitorM4. HoloMonitor M4 assists researchers to overcome inherent disadvantages of fluorescent analysis, specifically effects of chemical labels or genetic modifications which can alter cellular behavior. Additionally, label-free analysis is simple and eliminates the costs associated with staining procedures. The underlying technology principle is based on digital off-axis holography. While multiple alternatives exist for this type of analysis, we prioritized our developments to achieve the following: a) All-inclusive system - hardware and sophisticated cytometric analysis software; b) Ease of use enabling utilization of instrumentation by expert- and entrylevel researchers alike; c) Validated quantitative assay end-points tracked over time such as optical path length shift, optical volume and multiple derived imaging parameters; d) Reliable digital autofocus; e) Robust long-term operation in the incubator environment; f) High throughput and walk-away capability; and finally g) Data management suitable for single- and multi-user networks. We provide examples of HoloMonitor applications of label-free cell viability measurements and monitoring of cell cycle phase distribution.

  11. Real-Time, Label-Free Detection of Biomolecular Interactions in Sandwich Assays by the Oblique-Incidence Reflectivity Difference Technique

    Directory of Open Access Journals (Sweden)

    Yung-Shin Sun

    2014-12-01

    Full Text Available One of the most important goals in proteomics is to detect the real-time kinetics of diverse biomolecular interactions. Fluorescence, which requires extrinsic tags, is a commonly and widely used method because of its high convenience and sensitivity. However, in order to maintain the conformational and functional integrality of biomolecules, label-free detection methods are highly under demand. We have developed the oblique-incidence reflectivity difference (OI-RD technique for label-free, kinetic measurements of protein-biomolecule interactions. Incorporating the total internal refection geometry into the OI-RD technique, we are able to detect as low as 0.1% of a protein monolayer, and this sensitivity is comparable with other label-free techniques such as surface plasmon resonance (SPR. The unique advantage of OI-RD over SPR is no need for dielectric layers. Moreover, using a photodiode array as the detector enables multi-channel detection and also eliminates the over-time signal drift. In this paper, we demonstrate the applicability and feasibility of the OI-RD technique by measuring the kinetics of protein-protein and protein-small molecule interactions in sandwich assays.

  12. Self directed home based electrical muscle stimulation training improves exercise tolerance and strength in healthy elderly.

    Science.gov (United States)

    Caulfield, Brian; Prendergast, Ann; Rainsford, Gary; Minogue, Conor

    2013-01-01

    Advancing age is associated with a gradual decline in muscle strength, exercise tolerance and subsequent capacity for activities of daily living. It is important that we develop effective strategies to halt this process of gradual decline in order to enhance functional ability and capacity for independent living. To achieve this, we must overcome the challenge of sustaining ongoing engagement in physical exercise programmes in the sedentary elderly population, particularly those who experience barriers to exercise participation. Recent developments in electrical muscle stimulation technology could provide a potential solution. In this pilot case-control study we investigated the effects of a self-directed home based programme of electrical muscle stimulation training on muscle strength and exercise tolerance in a group of 16 healthy elderly volunteers (10f, 6m). Study participants completed 30 separate 1-hour electrical muscle stimulation sessions at home over a 6-week period. We observed significant improvements in quadriceps muscle strength and 6-minute walk distance, suggesting that this form of electrical muscle stimulation training has promise as an exercise modality in the elderly population.

  13. The direct writing and focusing of nanoparticles generated by an electrical discharge

    International Nuclear Information System (INIS)

    Saleh, E.; Praeger, M.; Vaughan, A. S.; Stewart, W.; Loh, W. H.

    2012-01-01

    Direct writing aims to deposit materials onto substrates in localised positions. In this paper, we demonstrate a new method for direct writing of nanoparticles at ambient-air-pressure. An electrical discharge is used to generate gold nanoparticles of the order of 10 nm diameter, which are then transported and ‘focused’ by an electric field in air, through the process of electric field-assisted diffusion, as opposed to normal ballistic focusing since the mean free path in air is very short. This process is novel and allows for practical normal atmospheric-pressure focused deposition of nanoparticles. The focusing mechanism is capable of producing patterned arrays of deposited nanoparticles with widths that are less than 10 % of the diameter of the focusing apparatus; in the present experimental configuration, gold spots with diameters of a few tens of micrometres were achieved, with ultimate size being limited by transverse diffusion and by charged particle mutual repulsion. In this study, the process of generating nanoparticles from bulk material, transporting and focusing these particles takes place in one operation, which is a key advantage in rapid prototyping and manufacturing techniques.

  14. Label-free detection of rheumatoid factor using YbY{sub x}O{sub y} electrolyte–insulator–semiconductor devices

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Tung-Ming, E-mail: tmpan@mail.cgu.edu.tw [Department of Electronics Engineering, Chang Gung University, Taoyuan 33302, Taiwan (China); Lin, Ting-Wei [School of Medicine, Chang Gung University, Taoyuan 33302, Taiwan (China); Chen, Ching-Yi [Department of Electronics Engineering, Chang Gung University, Taoyuan 33302, Taiwan (China)

    2015-09-03

    In this study, we investigated the effect of yttrium content on the structural properties and sensing characteristics of YbY{sub x}O{sub y} sensing membranes for electrolyte–insulator–semiconductor (EIS) sensors to detect the rheumatoid factor (RF). The YbY{sub x}O{sub y} EIS device prepared at the 60 W plasma condition exhibited a higher sensitivity of 65.77 mV/pH, a lower hysteresis voltage of ∼1 mV, and a smaller drift rate of 0.14 mV/h than did those prepared at the other conditions. We attribute this behavior to the optimal yttrium content in the YbY{sub x}O{sub y} film forming a smooth surface. Furthermore, we used a novel YbTi{sub x}O{sub y} EIS biosensor to measure the RF antigen in human serum because of its rapid and label-free detection. Two different techniques were used for the immobilization of RF antibody onto the surface of an YbTi{sub x}O{sub y} EIS sensor. The RF antibody was directly immobilized on the EIS surface modified with 3-aminopropyltriethoxysilane (APTES) followed by glutaraldehyde (GA). In contrast, a mixture of 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) solution was used to functionalize the carboxyl groups at the tail of RF antibodies. RF antibodies functionalized with the active NHS esters were covalently immobilized on the APTES-modified YbTi{sub x}O{sub y} surface. The immobilized RF antibodies on the EIS that are functionalized with the EDC and NHS exhibit higher (41.11 mV/pC{sub RF}) for detection of serum RF antigen in the range 10{sup −7} to 10{sup −3} M, compared to traditional antibody immobilization technique via APTES and GA linkage. The YbTi{sub x}O{sub y} EIS biosensor is a promising analytical tool for RF antigen monitoring due to its good sensitivity, stability and repeatability. - Highlights: • Effect of yttrium content on the structural and sensing properties of YbY{sub x}O{sub y} sensing membrane was explored. • YbY{sub x}O{sub y} EIS device prepared at the

  15. Electric Signals Regulate the Directional Migration of Oligodendrocyte Progenitor Cells (OPCs via β1 Integrin

    Directory of Open Access Journals (Sweden)

    Bangfu Zhu

    2016-11-01

    Full Text Available The guided migration of neural cells is essential for repair in the central nervous system (CNS. Oligodendrocyte progenitor cells (OPCs will normally migrate towards an injury site to re-sheath demyelinated axons; however the mechanisms underlying this process are not well understood. Endogenous electric fields (EFs are known to influence cell migration in vivo, and have been utilised in this study to direct the migration of OPCs isolated from neonatal Sprague-Dawley rats. The OPCs were exposed to physiological levels of electrical stimulation, and displayed a marked electrotactic response that was dependent on β1 integrin, one of the key subunits of integrin receptors. We also observed that F-actin, an important component of the cytoskeleton, was re-distributed towards the leading edge of the migrating cells, and that this asymmetric rearrangement was associated with β1 integrin function.

  16. In situ–Directed Growth of Organic Nanofibers and Nanoflakes: Electrical and Morphological Properties

    Directory of Open Access Journals (Sweden)

    de Oliveira Hansen Roana

    2011-01-01

    Full Text Available Abstract Organic nanostructures made from organic molecules such as para-hexaphenylene (p-6P could form nanoscale components in future electronic and optoelectronic devices. However, the integration of such fragile nanostructures with the necessary interface circuitry such as metal electrodes for electrical connection continues to be a significant hindrance toward their large-scale implementation. Here, we demonstrate in situ–directed growth of such organic nanostructures between pre-fabricated contacts, which are source–drain gold electrodes on a transistor platform (bottom-gate on silicon dioxide patterned by a combination of optical lithography and electron beam lithography. The dimensions of the gold electrodes strongly influence the morphology of the resulting structures leading to notably different electrical properties. The ability to control such nanofiber or nanoflake growth opens the possibility for large-scale optoelectronic device fabrication.

  17. Coordinated Operation of the Electricity and Natural Gas Systems with Bi-directional Energy Conversion

    DEFF Research Database (Denmark)

    Zeng, Qing; Zhang, Baohua; Fang, Jiakun

    2017-01-01

    A coordinated operation of the natural gas and electricity network with bi-directional energy conversion is expected to accommodate high penetration levels of renewables. This work focuses on the unified optimal operation of the integrated natural gas and electricity system considering the network...... constraints in both systems. An iterative method is proposed to deal with the nonlinearity in the proposed model. The models of the natural gas and power system are linearized in every iterative step. Simulation results demonstrate the effectiveness of the approach. Applicability of the proposed method...... is tested in the sample case. Finally, the effect of Power to Gas (P2G) on the daily economic dispatch is also investigated....

  18. Three-dimensional direct laser written graphitic electrical contacts to randomly distributed components

    Science.gov (United States)

    Dorin, Bryce; Parkinson, Patrick; Scully, Patricia

    2018-04-01

    The development of cost-effective electrical packaging for randomly distributed micro/nano-scale devices is a widely recognized challenge for fabrication technologies. Three-dimensional direct laser writing (DLW) has been proposed as a solution to this challenge, and has enabled the creation of rapid and low resistance graphitic wires within commercial polyimide substrates. In this work, we utilize the DLW technique to electrically contact three fully encapsulated and randomly positioned light-emitting diodes (LEDs) in a one-step process. The resolution of the contacts is in the order of 20 μ m, with an average circuit resistance of 29 ± 18 kΩ per LED contacted. The speed and simplicity of this technique is promising to meet the needs of future microelectronics and device packaging.

  19. Label-free viscosity measurement of complex fluids using reversal flow switching manipulation in a microfluidic channel.

    Science.gov (United States)

    Jun Kang, Yang; Ryu, Jeongeun; Lee, Sang-Joon

    2013-01-01

    The accurate viscosity measurement of complex fluids is essential for characterizing fluidic behaviors in blood vessels and in microfluidic channels of lab-on-a-chip devices. A microfluidic platform that accurately identifies biophysical properties of blood can be used as a promising tool for the early detections of cardiovascular and microcirculation diseases. In this study, a flow-switching phenomenon depending on hydrodynamic balancing in a microfluidic channel was adopted to conduct viscosity measurement of complex fluids with label-free operation. A microfluidic device for demonstrating this proposed method was designed to have two inlets for supplying the test and reference fluids, two side channels in parallel, and a junction channel connected to the midpoint of the two side channels. According to this proposed method, viscosities of various fluids with different phases (aqueous, oil, and blood) in relation to that of reference fluid were accurately determined by measuring the switching flow-rate ratio between the test and reference fluids, when a reverse flow of the test or reference fluid occurs in the junction channel. An analytical viscosity formula was derived to measure the viscosity of a test fluid in relation to that of the corresponding reference fluid using a discrete circuit model for the microfluidic device. The experimental analysis for evaluating the effects of various parameters on the performance of the proposed method revealed that the fluidic resistance ratio ( R J L / R L , fluidic resistance in the junction channel ( R J L ) to fluidic resistance in the side channel ( R L )) strongly affects the measurement accuracy. The microfluidic device with smaller R J L / R L values is helpful to measure accurately the viscosity of the test fluid. The proposed method accurately measured the viscosities of various fluids, including single-phase (Glycerin and plasma) and oil-water phase (oil vs. deionized water) fluids, compared with conventional

  20. The sheep (Ovis aries) muscle proteome: Decoding the mechanisms of tolerance to Seasonal Weight Loss using label-free proteomics.

    Science.gov (United States)

    Ferreira, Ana M; Grossmann, Jonas; Fortes, Claudia; Kilminster, Tanya; Scanlon, Tim; Milton, John; Greeff, Johan; Oldham, Chris; Nanni, Paolo; Almeida, André M

    2017-05-24

    Seasonal Weight Loss (SWL) is one of the most pressing issues in animal production in the tropics and Mediterranean. This work aims to characterize muscle proteome changes as a consequence of SWL in meat producing sheep, using a label-free proteomics approach. We compare three breeds: the Australian Merino (SWL susceptible), the Damara (SWL tolerant) and the Dorper (SWL intermediate tolerance). We identified 668 proteins of the sheep proteome, 95 with differential regulation. Also we observe that the more vulnerable to SWL a breed is, the more differential abundance proteins we find. Protein binding was the most frequently altered molecular function identified. We suggest 6 putative markers for restricted nutritional conditions independently of breed: ferritin heavy-chain; immunoglobulin V lambda chain; transgelin; fatty acid synthase; glutathione S-transferase A2; dihydrodiol dehydrogenase 3-like. Moreover, we suggest as related to SWL tolerance: S100-A10 Serpin A3-5-like and Catalase, subject however to necessary validation assays. The identification of SWL-tolerance related proteins using proteomics will lead to increased stock productivity of relevant interest to animal production, particularly if identified at the muscle level, the tissue of economic importance in meat production. Seasonal Weight Loss (SWL) is the most pressing issue in animal production in the tropics and the Mediterranean. To counter SWL, farmers often use animal breeds that have a natural ability to withstand pasture scarcity. Here we study the sheep muscle proteome at the muscle level, the tissue of economic importance in meat production. Furthermore, the identification of proteins that change their abundance in response to SWL using proteomics can contribute to increased stock productivity of relevant interest to animal production. We identified 668 proteins of the sheep proteome. We demonstrate that the following proteins are affected by restricted nutritional conditions: ferritin heavy

  1. Targeted proteomics guided by label-free global proteome analysis in saliva reveal transition signatures from health to periodontal disease.

    Science.gov (United States)

    Bostanci, Nagihan; Selevsek, Nathalie; Wolski, Witold; Grossmann, Jonas; Bao, Kai; Wahlander, Asa; Trachsel, Christian; Schlapbach, Ralph; Özturk, Veli Özgen; Afacan, Beral; Emingil, Gulnur; Belibasakis, Georgios N

    2018-04-02

    Periodontal diseases are among the most prevalent worldwide, but largely silent, chronic diseases. They affect the tooth-supporting tissues with multiple ramifications on life quality. Their early diagnosis is still challenging, due to lack of appropriate molecular diagnostic methods. Saliva offers a non-invasively collectable reservoir of clinically relevant biomarkers, which, if utilized efficiently, could facilitate early diagnosis and monitoring of ongoing disease. Despite several novel protein markers being recently enlisted by discovery proteomics, their routine diagnostic application is hampered by the lack of validation platforms that allow for rapid, accurate and simultaneous quantification of multiple proteins in large cohorts. We carried out a pipeline of two proteomic platforms; firstly, we applied open ended label-free quantitative (LFQ) proteomics for discovery in saliva (n=67, health, gingivitis, and periodontitis), followed by selected-reaction monitoring (SRM)-targeted proteomics for validation in an independent cohort (n=82). The LFQ platform led to the discovery of 119 proteins with at least two-fold significant difference between health and disease. The 65 proteins chosen for the subsequent SRM platform included 50 related proteins derived from the significantly enriched processes of the LFQ data, 11 from literature-mining, and four house-keeping ones. Among those, 60 were reproducibly quantifiable proteins (92% success rate), represented by a total of 143 peptides. Machine-learning modeling led to a narrowed-down panel of five proteins of high predictive value for periodontal diseases (higher in disease: Matrix metalloproteinase-9, Ras-related protein-1, Actin-related protein 2/3 complex subunit 5; lower in disease: Clusterin, Deleted in Malignant Brain Tumors 1), with maximum area under the receiver operating curve >0.97. This panel enriches the pool of credible clinical biomarker candidates for diagnostic assay development. Yet, the quantum

  2. Electroosmotic velocity and electric conductivity in a fibrous porous medium in the transverse direction.

    Science.gov (United States)

    Keh, Huan J; Wu, Yi Y

    2011-07-28

    The steady electroosmosis and electric conduction in a fibrous medium constructed by a homogeneous array of parallel, identical, charged, circular cylinders filled with an electrolyte solution is analytically examined. The imposed electric field is constant and normal to the axes of the cylinders. The electric double layer surrounding each dielectric cylinder may have an arbitrary thickness relative to the radius of the cylinder. A unit cell model that allows for the overlap of the double layers of adjacent cylinders is employed. The electrokinetic equations that govern the ionic concentration distributions, the electrostatic potential profile, and the fluid flow field in the electrolyte solution surrounding the charged cylinder in a cylindrical cell are linearized assuming that the system is only slightly distorted from equilibrium. Through the use of a regular perturbation method, these linearized equations are solved with the surface charge density (or zeta potential) of the cylinder as the small perturbation parameter. Analytical expressions for the electroosmotic velocity of the fluid solution and the effective electric conductivity in the array of cylinders are obtained in closed forms as functions of the porosity of the fiber matrix and other characteristics of the porous system. Comparisons of the results of the cell model with different conditions at the outer boundary of the cell are made. The cell model predicts that, under otherwise identical conditions, the electric conductivity in a porous medium composed of an array of parallel cylinders in the transverse direction in general is smaller than that of a suspension of spheres, but there are some exceptions. The effect of interactions among the cylinders or spheres on the effective conductivity can be significant under appropriate conditions. © 2011 American Chemical Society

  3. Ultrasensitive label-free detection of DNA hybridization by sapphire-based graphene field-effect transistor biosensor

    Science.gov (United States)

    Xu, Shicai; Jiang, Shouzhen; Zhang, Chao; Yue, Weiwei; Zou, Yan; Wang, Guiying; Liu, Huilan; Zhang, Xiumei; Li, Mingzhen; Zhu, Zhanshou; Wang, Jihua

    2018-01-01

    Graphene has attracted much attention in biosensing applications for its unique properties. Because of one-atom layer structure, every atom of graphene is exposed to the environment, making the electronic properties of graphene are very sensitive to charged analytes. Therefore, graphene is an ideal material for transistors in high-performance sensors. Chemical vapor deposition (CVD) method has been demonstrated the most successful method for fabricating large area graphene. However, the conventional CVD methods can only grow graphene on metallic substrate and the graphene has to be transferred to the insulating substrate for further device fabrication. The transfer process creates wrinkles, cracks, or tears on the graphene, which severely degrade electrical properties of graphene. These factors severely degrade the sensing performance of graphene. Here, we directly fabricated graphene on sapphire substrate by high temperature CVD without the use of metal catalysts. The sapphire-based graphene was patterned and make into a DNA biosensor in the configuration of field-effect transistor. The sensors show high performance and achieve the DNA detection sensitivity as low as 100 fM (10-13 M), which is at least 10 times lower than prior transferred CVD G-FET DNA sensors. The use of the sapphire-based G-FETs suggests a promising future for biosensing applications.

  4. An energy management system for a directly-driven electric scooter

    International Nuclear Information System (INIS)

    Yang, Yee-Pien; Liu, Jieng-Jang; Hu, Tsung-Hsien

    2011-01-01

    An energy management system with an electronic gearshift and regenerative braking is presented to improve the gross efficiency and driving range of an electric scooter, driven directly by a four-phase axial-flux DC brushless wheel motor. The integration of stator windings, batteries, ultracapacitors, and a digital controller constitutes an energy management system, which features smooth electronic gear shifting and regenerative braking. The gross efficiency of the experimental scooter is improved in the drivable range by 20% with respect to that without regenerative braking. The battery-to-wheel efficiency was also above 70% for both low- and high-speed gears.

  5. Electrical Thermal Network for Direct Contact Membrane Distillation Modeling and Analysis

    KAUST Repository

    Karam, Ayman M.

    2015-02-04

    Membrane distillation is an emerging water distillation technology that offers several advantages compared to conventional water desalination processes. Although progress has been made to model and understand the physics of the process, many studies are based on steady-state assumptions or are computationally not appropriate for real time control. This paper presents the derivation of a novel dynamical model, based on analogy between electrical and thermal systems, for direct contact membrane distillation (DCMD). The proposed model captures the dynamics of temperature distribution and distilled water flux. To demonstrate the adequacy of the proposed model, validation with transient and steady-state experimental data is presented.

  6. Simulation of LOF accidents with directly electrical heated UO2 pins

    International Nuclear Information System (INIS)

    Alexas, A.

    1976-01-01

    The behavior of directly electrical heated UO 2 pins has been investigated under loss of coolant conditions. Two types of hypothetical accidents have been simulated, first, a LOF accident without power excursion (LOF accident) and second, a LOF accident with subsequent power excursion (LOF-TOP accident). A high-speed film shows the sequence of events for two characteristic experiments. In consequence of the high-speed film analysis as well as the metallographical evaluation statements are given in respect to the cladding meltdown process, the fuel melt fraction and the energy input from the beginning of a power transient to the beginning of the molten fuel ejections

  7. Atomistic modeling of metal surfaces under electric fields: direct coupling of electric fields to a molecular dynamics algorithm

    CERN Document Server

    Djurabekova, Flyura; Pohjonen, Aarne; Nordlund, Kai

    2011-01-01

    The effect of electric fields on metal surfaces is fairly well studied, resulting in numerous analytical models developed to understand the mechanisms of ionization of surface atoms observed at very high electric fields, as well as the general behavior of a metal surface in this condition. However, the derivation of analytical models does not include explicitly the structural properties of metals, missing the link between the instantaneous effects owing to the applied field and the consequent response observed in the metal surface as a result of an extended application of an electric field. In the present work, we have developed a concurrent electrodynamic–molecular dynamic model for the dynamical simulation of an electric-field effect and subsequent modification of a metal surface in the framework of an atomistic molecular dynamics (MD) approach. The partial charge induced on the surface atoms by the electric field is assessed by applying the classical Gauss law. The electric forces acting on the partially...

  8. Cytotoxicity, Bactericidal, and Antioxidant Activity of Sodium Alginate Hydrosols Treated with Direct Electric Current.

    Science.gov (United States)

    Król, Żaneta; Marycz, Krzysztof; Kulig, Dominika; Marędziak, Monika; Jarmoluk, Andrzej

    2017-03-22

    The aim of the study was to investigate the effect of using direct electric current (DC) of 0, 200, and 400 mA for five minutes on the physiochemical properties, cytotoxicity, antibacterial, and antioxidant activity of sodium alginate hydrosols with different sodium chloride concentrations. The pH, oxidation-reduction potential (ORP), electrical conductivity (EC), and available chlorine concentration (ACC) were measured. The effect of sodium alginate hydrosols treated with DC on Staphylococcus aureus , Listeria monocytogenes , Bacillus cereus , Micrococcus luteus , Escherichia coli , Salmonella enteritidis , Yersinia enterocolitica , Pseudomonas fluorescence , and RAW 264.7 and L929 cells was investigated. Subsequently, the antioxidant properties of hydrosols were evaluated by determining the scavenging ability of 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH) and ferric reducing antioxidant power (FRAP). The results have shown that after applying 400 mA in hydrosol samples with 0.1% and 0.2% NaCl all tested bacteria were inactivated. The ACC concentration of C400 samples with NaCl was equal to 13.95 and 19.71 mg/L, respectively. The cytotoxicity analysis revealed that optimized electric field conditions and the addition of sodium chloride allow for the avoidance of toxicity effects on normal cells without disturbing the antibacterial effects. Due to the presence of oxidizing substances, the DPPH of variants treated with DC was lower than the DPPH of control samples.

  9. Optical gain in colloidal quantum dots achieved with direct-current electrical pumping

    Science.gov (United States)

    Lim, Jaehoon; Park, Young-Shin; Klimov, Victor I.

    2018-01-01

    Chemically synthesized semiconductor quantum dots (QDs) can potentially enable solution-processable laser diodes with a wide range of operational wavelengths, yet demonstrations of lasing from the QDs are still at the laboratory stage. An important challenge--realization of lasing with electrical injection--remains unresolved, largely due to fast nonradiative Auger recombination of multicarrier states that represent gain-active species in the QDs. Here we present population inversion and optical gain in colloidal nanocrystals realized with direct-current electrical pumping. Using continuously graded QDs, we achieve a considerable suppression of Auger decay such that it can be outpaced by electrical injection. Further, we apply a special current-focusing device architecture, which allows us to produce high current densities (j) up to ~18 A cm-2 without damaging either the QDs or the injection layers. The quantitative analysis of electroluminescence and current-modulated transmission spectra indicates that with j = 3-4 A cm-2 we achieve the population inversion of the band-edge states.

  10. Electrically conductive epoxy nanocomposites with expanded graphite/carbon nanotube hybrid fillers prepared by direct hybridization.

    Science.gov (United States)

    Yu, Lan; Kang, Hyokyung; Lim, Yun-Soo; Lee, Churl Seung; Shin, Kwonwoo; Park, Ji Sun; Han, Jong Hun

    2014-12-01

    Carbon nanotubes (CNTs) are generally used to promote the electrical conductivity of the polymer nanocomposites. However, in spite of their superior properties, CNT's high cost has limited their commercial application, so far. Thus, the development of hybrid carbon nanomaterials (CNMs) composed of CNTs and cheaper CNMs such as carbon fibers (CFs), expanded graphites (EGs), and graphene nanoplatelets (GNPs) is important in terms of reducing the cost of CNT-based fillers. In this study, we prepared EG/CNT hybrid fillers via direct CNT synthesis on the EG support using modified combustion method and thermal chemical vapor deposition (CVD) method, and investigated the electrical conductivity of the expoxy nanocomposite with EG/CNT hybrid fillers. The epoxy nanocomposites with EG/CNT hybrid fillers at 20 wt% filler loading showed 260% and 170% electrical conductivity enhancement in comparison with the EG and the simply mixed EG and CNT fillers, respectively. Our approach provides various applications including electromagnetic interference (EMI) shielding materials, thermal interface materials (TIMs), and reinforced nanocomposites.

  11. Directed Field Ionization: A Genetic Algorithm for Evolving Electric Field Pulses

    Science.gov (United States)

    Kang, Xinyue; Rowley, Zoe A.; Carroll, Thomas J.; Noel, Michael W.

    2017-04-01

    When an ionizing electric field pulse is applied to a Rydberg atom, the electron's amplitude traverses many avoided crossings among the Stark levels as the field increases. The resulting superposition determines the shape of the time resolved field ionization spectrum at a detector. An engineered electric field pulse that sweeps back and forth through avoided crossings can control the phase evolution so as to determine the electron's path through the Stark map. In the region of n = 35 in rubidium there are hundreds of potential avoided crossings; this yields a large space of possible pulses. We use a genetic algorithm to search this space and evolve electric field pulses to direct the ionization of the Rydberg electron in rubidium. We present the algorithm along with a comparison of simulated and experimental results. This work was supported by the National Science Foundation under Grants No. 1607335 and No. 1607377 and used the Extreme Science and Engineering Discovery Environment (XSEDE), which is supported by National Science Foundation Grant Number OCI-1053575.

  12. Cytotoxicity, Bactericidal, and Antioxidant Activity of Sodium Alginate Hydrosols Treated with Direct Electric Current

    Directory of Open Access Journals (Sweden)

    Żaneta Król

    2017-03-01

    Full Text Available The aim of the study was to investigate the effect of using direct electric current (DC of 0, 200, and 400 mA for five minutes on the physiochemical properties, cytotoxicity, antibacterial, and antioxidant activity of sodium alginate hydrosols with different sodium chloride concentrations. The pH, oxidation-reduction potential (ORP, electrical conductivity (EC, and available chlorine concentration (ACC were measured. The effect of sodium alginate hydrosols treated with DC on Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Micrococcus luteus, Escherichia coli, Salmonella enteritidis, Yersinia enterocolitica, Pseudomonas fluorescence, and RAW 264.7 and L929 cells was investigated. Subsequently, the antioxidant properties of hydrosols were evaluated by determining the scavenging ability of 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH and ferric reducing antioxidant power (FRAP. The results have shown that after applying 400 mA in hydrosol samples with 0.1% and 0.2% NaCl all tested bacteria were inactivated. The ACC concentration of C400 samples with NaCl was equal to 13.95 and 19.71 mg/L, respectively. The cytotoxicity analysis revealed that optimized electric field conditions and the addition of sodium chloride allow for the avoidance of toxicity effects on normal cells without disturbing the antibacterial effects. Due to the presence of oxidizing substances, the DPPH of variants treated with DC was lower than the DPPH of control samples.

  13. 75 FR 70096 - Airworthiness Directives; General Electric Company (GE) CT7-9C and -9C3 Turboprop Engines

    Science.gov (United States)

    2010-11-17

    ... Airworthiness Directives; General Electric Company (GE) CT7-9C and -9C3 Turboprop Engines AGENCY: Federal... published life limit, and which could result in an uncontained engine failure and damage to the airplane... service information identified in this AD, contact General Electric Company, GE-Aviation, Room 285, 1...

  14. Directional Congestion and Regime Switching in a Long Memory Model for Electricity Prices

    DEFF Research Database (Denmark)

    Haldrup, Niels; Nielsen, Morten Ø.

    The functioning of electricity markets has experienced increasing complexityas a result of deregulation in recent years. Consequently this affects the multilateral price behaviour across regions with physical exchange of power. It has been documented elsewhere that features such aslong memory...... and regime switching reflecting congestion and non-congestion periods are empirically relevant and hence are features that need to be taken into account when modeling price behavior. In the present paper we further elaborate on the co-existence of long memory and regime switches by focusing on the effect...... that the direction of possible congestion episodes has on the price dynamics. Under non-congestion prices are identical. The direction of possible congestion is identified by the region with excess demand of power through the sign of price differences and hence three different states can be considered: Non...

  15. Electrical equivalent thermal network for direct contact membrane distillation modeling and analysis

    KAUST Repository

    Karam, Ayman M.

    2016-09-19

    Membrane distillation (MD) is an emerging water desalination technology that offers several advantages compared to conventional desalination methods. Although progress has been made to model the physics of the process, there are two common limitations of existing models. Firstly, many of the models are based on the steady-state analysis of the process and secondly, some of the models are based on partial differential equations, which when discretized introduce many states which are not accessible in practice. This paper presents the derivation of a novel dynamic model, based on the analogy between electrical and thermal systems, for direct contact membrane distillation (DCMD). An analogous electrical thermal network is constructed and its elements are parameterized such that the response of the network models the DCMD process. The proposed model captures the spatial and temporal responses of the temperature distribution along the flow direction and is able to accurately predict the distilled water flux output. To demonstrate the adequacy of the proposed model, validation with time varying and steady-state experimental data is presented. (C) 2016 Elsevier Ltd. All rights reserved.

  16. Direct prediction of spatially and temporally varying physical properties from time-lapse electrical resistance data

    Science.gov (United States)

    Hermans, Thomas; Oware, Erasmus; Caers, Jef

    2016-09-01

    Time-lapse applications of electrical methods have grown significantly over the last decade. However, the quantitative interpretation of tomograms in terms of physical properties, such as salinity, temperature or saturation, remains difficult. In many applications, geophysical models are transformed into hydrological models, but this transformation suffers from spatially and temporally varying resolution resulting from the regularization used by the deterministic inversion. In this study, we investigate a prediction-focused approach (PFA) to directly estimate subsurface physical properties with electrical resistance data, circumventing the need for classic tomographic inversions. First, we generate a prior set of resistance data and physical property forecast through hydrogeological and geophysical simulations mimicking the field experiment. We reduce the dimension of both the data and the forecast through principal component analysis in order to keep the most informative part of both sets in a reduced dimension space. Then, we apply canonical correlation analysis to explore the relationship between the data and the forecast in their reduced dimension space. If a linear relationship can be established, the posterior distribution of the forecast can be directly sampled using a Gaussian process regression where the field data scores are the conditioning data. In this paper, we demonstrate PFA for various physical property distributions. We also develop a framework to propagate the estimated noise level in the reduced dimension space. We validate the results by a Monte Carlo study on the posterior distribution and demonstrate that PFA yields accurate uncertainty for the cases studied.

  17. Field emission driven direct current argon discharges and electrical breakdown mechanism across micron scale gaps

    Science.gov (United States)

    Matejčik, Štefan; Radjenović, Branislav; Klas, Matej; Radmilović-Radjenović, Marija

    2015-11-01

    In this paper results of the experimental and theoretical studies of the field emission driven direct current argon microdischarges for the gaps between 1 μm and 100 μm are presented and discussed. The breakdown voltage curves and Volt-Ampere characteristics proved to be a fertile basis providing better understanding of the breakdown phenomena in microgaps. Based on the measured breakdown voltage curves, the effective yields have been estimated confirming that the secondary electron emission due to high electric field generated in microgaps depends primarily on the electric field leading directly to the violation of the Paschen's law. Experimental data are supported by the theoretical predictions that suggest departure from the scaling law and a flattening of the Paschen curves at higher pressures confirming that Townsend phenomenology breaks down when field emission becomes the key mechanism leading to the breakdown. Field emission of electrons from the cathode, the space charge effects in the breakdown and distinction between the Fowler-Nordheim field emission and the space charge limited current density are also analyzed. Images and Volt-Ampere characteristics recorded at the electrode gap size of 20 μm indicate the existence of a discharge region similar to arc at the pressure of around 200 Torr has been observed. Contribution to the Topical Issue "Recent Breakthroughs in Microplasma Science and Technology", edited by Kurt Becker, Jose Lopez, David Staack, Klaus-Dieter Weltmann and Wei Dong Zhu.

  18. High-performance liquid-catalyst fuel cell for direct biomass-into-electricity conversion.

    Science.gov (United States)

    Liu, Wei; Mu, Wei; Deng, Yulin

    2014-12-01

    Herein, we report high-performance fuel cells that are catalyzed solely by polyoxometalate (POM) solution without any solid metal or metal oxide. The novel design of the liquid-catalyst fuel cells (LCFC) changes the traditional gas-solid-surface heterogeneous reactions to liquid-catalysis reactions. With this design, raw biomasses, such as cellulose, starch, and even grass or wood powders can be directly converted into electricity. The power densities of the fuel cell with switchgrass (dry powder) and bush allamanda (freshly collected) are 44 mW cm(-2) and 51 mW cm(-2) respectively. For the cellulose-based biomass fuel cell, the power density is almost 3000 times higher than that of cellulose-based microbial fuel cells. Unlike noble-metal catalysts, POMs are tolerant to most organic and inorganic contaminants. Therefore, almost any raw biomass can be used directly to produce electricity without prior purification. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Investigation of students’ intermediate conceptual understanding levels: the case of direct current electricity concepts

    International Nuclear Information System (INIS)

    Aktan, D Cobanoglu

    2013-01-01

    Conceptual understanding is one of the main topics in science and physics education research. In the majority of conceptual understanding studies, students’ understanding levels were categorized dichotomously, either as alternative or scientific understanding. Although they are invaluable in many ways, namely developing new instructional materials and assessment instruments, students’ alternative understandings alone are not sufficient to describe students’ conceptual understanding in detail. This paper introduces an example of a study in which a method was developed to assess and describe students’ conceptual understanding beyond alternative and scientific understanding levels. In this study, six undergraduate students’ conceptual understanding levels of direct current electricity concepts were assessed and described in detail by using their answers to qualitative problems. In order to do this, conceptual understanding indicators are described based on science and mathematics education literature. The students’ understanding levels were analysed by assertion analysis based on the conceptual understanding indicators. The results indicated that the participants demonstrated three intermediate understanding levels in addition to alternative and scientific understanding. This paper presents the method and its application to direct current electricity concepts. (paper)

  20. Label-free quantitative proteomic analysis of pre-flowering PMeV-infected Carica papaya L.

    Science.gov (United States)

    Soares, Eduardo de A; Werth, Emily G; Madroñero, Leidy J; Ventura, José A; Rodrigues, Silas P; Hicks, Leslie M; Fernandes, Patricia M B

    2017-01-16

    Papaya meleira virus (PMeV) infects papaya (Carica papaya L.) and leads to Papaya Sticky Disease (PSD) or "Meleira", characterized by a spontaneous exudation of latex from fruits and leaves only in the post-flowering developmental stage. The latex oxidizes in contact with air and accumulates as a sticky substance on the plant organs, impairing papaya fruit's marketing and exportation. To understand pre-flowering C. papaya resistance to PMeV, an LC-MS/MS-based label-free proteomics approach was used to assess the differential proteome of PMeV-infected pre-flowering C. papaya vs. uninfected (control) plants. In this study, 1333 proteins were identified, of which 111 proteins showed a significant abundance change (57 increased and 54 decreased) and supports the hypothesis of increased photosynthesis and reduction of 26S-proteassoma activity and cell-wall remodeling. All of these results suggest that increased photosynthetic activity has a positive effect on the induction of plant immunity, whereas the reduction of caspase-like activity and the observed changes in the cell-wall associated proteins impairs the full activation of defense response based on hypersensitive response and viral movement obstruction in pre-flowering C. papaya plants. The papaya (Carica papaya L.) fruit's production is severely limited by the occurrence of Papaya meleira virus (PMeV) infection, which causes Papaya Sticky Disease (PSD). Despite the efforts to understand key features involved with the plant×virus interaction, PSD management is still largely based on the observation of the first disease symptoms in the field, followed by the elimination of the diseased plants. However, C. papaya develops PSD only after flowering, i.e. about six-months after planting, and the virus inoculum sources are kept in field. The development of PMeV resistant genotypes is impaired by the limited knowledge about C. papaya resistance against viruses. The occurrence of a resistance/tolerance mechanism to PSD