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Book Study of Early Signaling Events in T Cell Activation Enabled Through a Modular and Multi time Point Microfluidic Device

Download or read book Study of Early Signaling Events in T Cell Activation Enabled Through a Modular and Multi time Point Microfluidic Device written by Catherine Aurelie Rivet and published by . This book was released on 2008 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Binding of the antigen receptor on T cells initiates a rapid series of signaling events leading to an immune response. To fully understand T cell mediated immunity, underlying regulatory properties of the receptor network must be understood. Monitoring dynamic protein signaling events allows for network analysis. Unfortunately, dynamic data acquisition is often extremely time-consuming and expensive with conventional methods; the number of proteins monitored at the same time on the same sample is limited. Furthermore, with conventional, multi-well plate assays it is difficult to achieve adequate resolution at sub-minute timescales. Microfluidics is a capable alternative, providing uniformity in sample handling to reduce error between experiments and precision in timing, an important factor in monitoring phosphorylation events that occur within minutes of stimulation. We used a two-module microfluidic platform for simultaneous multi-time point stimulation and lysis of T cells to investigate early signaling events with a resolution down to 20 seconds using only small amounts of cells and reagents. The device did not elicit adverse cellular stress in Jurkat cells. The activation of 6 important proteins in the signaling cascade upon stimulation with a soluble form of Î"--CD3 in the device was quantified and compared under a variety of conditions. First, in comparison to manual pipetting, the microdevice exhibits significantly less error between experiments. Secondly, a comparison between Jurkat cells and primary T cells shows similar dynamic trends across the 6 proteins. Finally, we have used the device to compare properties of long-term vs, short-term cultured primary T cells. As expected, older cells present a much weakened response to antigenic cues, as measured with TCR response markers. This modular microdevice provides a flexible format for investigating cell signaling properties through the use of soluble cue stimuli.

Book Interrogating Immune Signaling at Single cell Resolution Using Microfluidic Systems

Download or read book Interrogating Immune Signaling at Single cell Resolution Using Microfluidic Systems written by Meiye Wu and published by . This book was released on 2012 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Mammalian cell populations are heterogeneous and plastic in their response to stimuli and communication with each other. Multi-parameter quantitative experimental and computational methods that preserve single-cell resolution in these complex cell populations are necessary for elucidating the highly dynamic mechanisms that underlie cell signaling. This thesis explores various aspects of immune cell signaling using integrated microfluidic lab-on-a-chip technologies, capable of generating such multi-parameter single-cell measurements. A brief survey of existing single-cell resolution techniques and their applications in cell signaling studies, including microfluidic devices that evolved into those used in this thesis is discussed. Three microfluidic methods are developed for studying innate and adaptive immune cell signaling. The first method enables the spatiotemporal profiling of the toll-like receptor 4 (TLR4) pathway using a microfluidic device that seamlessly integrates cell culture, sample preparation, imaging and flow cytometry capabilities. Systems-level profiling of signaling proteins in the TLR4 pathway is performed in a single experiment on the integrated microfluidic platform, to generate data used for subsequent mathematical modeling. Next, a new single-cell resolution microfluidic method that combines metabolic labeling of bioorthogonal probes and proximity ligation assay to measure dynamically glycosylated proteins, a post-translational modification with widespread significance in cell signaling modulation, is demonstrated in T cells. The third method extends the utility of the microfluidic platform to measure microRNAs concurrently with proteins in intact T cells, to further expand the repertoire of signaling molecules that can be multiplexed and quantified at single-cell resolution. The lab-on-a-chip technologies developed in this thesis present potential uses for several important applications including basic research, drug and biomarker discovery, biodefense, and for development of cost-effective point-of-care clinical diagnostics.

Book Leveraging Open Microfluidics for Platform Development and Cell signaling Studies In Vitro

Download or read book Leveraging Open Microfluidics for Platform Development and Cell signaling Studies In Vitro written by Samuel Berry and published by . This book was released on 2020 with total page 174 pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation discusses the fundamentals, development, validation, and application of open microfluidic technologies as a research tool in biological studies. Open microfluidics is a rapidly evolving and expanding field, characterized by the study of fluid behavior in channels with dimensions

Book Development of a Two phase Microfluidic Platform for Drug Screening

Download or read book Development of a Two phase Microfluidic Platform for Drug Screening written by Jenifer Clausell-Tormos and published by . This book was released on 2010 with total page 288 pages. Available in PDF, EPUB and Kindle. Book excerpt: High-throughput cell-based assays require small sam pie volumes to reduce assay costs and to allow for rapid sample manipulation. However, further miniaturization of conventional microtiter plate technology is problematic due to evaporation and capillary action. To overcome these limitations, we have developed a two-phase microfluidic platform in which human cells and multicellular organisms can be cultivated for several days in aqueous microcomparments separated by an inert perfluorocarbon carrier oil. Furthermore, we focused on the automated generation of chemically-dictinct microcompartment to exploit the technology for screening purposes. ln particular, we interfaced an autosampler with our microfluidic platform sequentially loading compounds from microtiter plates into a length of tubing. Ali compounds are loaded in form of aqueous plugs (nanoliter volumes) separated by fluorinated oil. The resulting array of plugs can be split into multiple small volume copies which can be used as replicates for the same assay as weil as for different assays. Moreover, each array of plugs can be injected into a microfluidic chip for further manipulation. Since the order of the compounds and thus their identity is known throughout the whole screening procedure, the system does not require direct compound labelling. Furthermore, each individual plug can be monitored over time, thus allowing the recording of kinetic data. In the last part of the work we focussed on the development of a novel assay coupling a positive fluorescence signal with the inhibition of viral transduction. This should ultimately allow the screening of antivirals in the previously developed microfluidic systems.

Book A Microfluidic Platform for the Genome wide Analysis of Electrical Phenotype

Download or read book A Microfluidic Platform for the Genome wide Analysis of Electrical Phenotype written by Michael David Vahey and published by . This book was released on 2010 with total page 143 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis presents the development of a new microfluidic method for separating and characterizing cells based upon differences in their electrical properties. The objective of this work is to obtain a genome-wide mapping of genotype to electrical phenotype in the budding yeast Saccharomyces cerevisiae. Towards this end, we present: (1) the development of a novel equilibrium gradient separation method, called isodielectric separation (IDS); (2) the development of physical theories describing how interactions between particles effect microscale separations; and (3) the application of IDS to a screen for electrical phenotypes in the yeast deletion library. Despite the variety of technologies available for cell sorting, a relative lack of intrinsic separation methods - those which separate cells according to their natural, unmodified characteristics - persists. To address this need, we have developed isodielectric separation. IDS separates cells according to differences in their intrinsic electrical properties. Using dielectrophoresis in a medium with spatially varying electrical conductivity, IDS drives cells to the locations where their polarization charges vanish, spatially resolving cells with different electrical properties. Our implementation of IDS offers label-free, continuous-flow separation, and is capable of resolving graded differences in electrical properties. Additionally, we demonstrate the ability to extract quantitative information from samples during separation, establishing IDS as an analytical technique as well as a preparative one. Any platform for performing genetic screens must have high throughput. Although satisfying this requirement would be greatly facilitated by using high cell densities, physical interactions between cells under these conditions can affect the performance of devices used for screening. Although pervasive, interactions between cells or particles are challenging to describe quantitatively, especially in the confined environments typical of microfluidic devices. By studying the effects of electrostatic and hydrodynamic interactions between particles in a microfluidic device, we have found that ensembles of interacting particles exhibit emergent behaviors that we are able to predict through numerical simulations and a simple analytic model based on hydrodynamic coupling. Applying these theoretical tools to IDS and other microfluidic separation methods has provided insight into how particle interactions can profoundly influence separation performance in counterintuitive ways. Having demonstrated the performance metrics necessary for a genetic screen, we apply IDS to the genome-wide analysis of electrical properties in the budding yeast S. cerevisiae. Although others have studied changes in electrical properties induced by drastic changes in gene expression (e.g. in differentiation) or by specific mutations in a small number of genes, a systematic and comprehensive analysis of the relationship between genotype and electrical phenotype has yet to be performed. Using IDS, we have screened the -5000 strains in the yeast deletion collection for altered electrical phenotype. This work has identified a number of genes associated with distinct electrical properties, and, by analyzing known interactions and correlations between these genes, we have identified pathways and morphologies that appear to be primary determinants of electrical phenotype.

Book Microfluidics for Single Cell Analysis

Download or read book Microfluidics for Single Cell Analysis written by Jin-Ming Lin and published by Springer Nature. This book was released on 2019-08-28 with total page 263 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book summarizes the various microfluidic-based approaches for single-cell capture, isolation, manipulation, culture and observation, lysis, and analysis. Single-cell analysis reveals the heterogeneities in morphology, functions, composition, and genetic performance of seemingly identical cells, and advances in single-cell analysis can overcome the difficulties arising due to cell heterogeneity in the diagnostics for a targeted model of disease. This book provides a detailed review of the state-of-the-art techniques presenting the pros and cons of each of these methods. It also offers lessons learned and tips from front-line investigators to help researchers overcome bottlenecks in their own studies. Highlighting a number of techniques, such as microfluidic droplet techniques, combined microfluidics-mass-spectrometry systems, and nanochannel sampling, it describes in detail a new microfluidic chip-based live single-cell extractor (LSCE) developed in the editor’s laboratory, which opens up new avenues to use open microfluidics in single-cell extraction, single-cell mass spectrometric analysis, single-cell adhesion analysis and subcellular operations. Serving as both an elementary introduction and advanced guidebook, this book interests and inspires scholars and students who are currently studying or wish to study microfluidics-based cell analysis methods.

Book Molecular Biology of B Cells

Download or read book Molecular Biology of B Cells written by Tasuku Honjo and published by Academic Press. This book was released on 2014-12-22 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular Biology of B Cells, Second Edition is a comprehensive reference to how B cells are generated, selected, activated and engaged in antibody production. All of these developmental and stimulatory processes are described in molecular, immunological, and genetic terms to give a clear understanding of complex phenotypes. Molecular Biology of B Cells, Second Edition offers an integrated view of all aspects of B cells to produce a normal immune response as a constant, and the molecular basis of numerous diseases due to B cell abnormality. The new edition continues its success with updated research on microRNAs in B cell development and immunity, new developments in understanding lymphoma biology, and therapeutic targeting of B cells for clinical application. With updated research and continued comprehensive coverage of all aspects of B cell biology, Molecular Biology of B Cells, Second Edition is the definitive resource, vital for researchers across molecular biology, immunology and genetics.

Book Biochip Technology

    Book Details:
  • Author : Jing Cheng
  • Publisher : CRC Press
  • Release : 2003-09-02
  • ISBN : 0203305043
  • Pages : 453 pages

Download or read book Biochip Technology written by Jing Cheng and published by CRC Press. This book was released on 2003-09-02 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biochip technology has experienced explosive growth in recent years and Biochip technology describes the basic manufacturing and fabrication processes and the current range of applications of these chips. Top scientists from the biochip industry and related areas explain the diverse applications of biochips in gene sequencing, expression monitoring, disease diagnosis, tumor examination, ligand assay and drug discovery.

Book Bioelectrics

Download or read book Bioelectrics written by Hidenori Akiyama and published by Springer. This book was released on 2016-09-19 with total page 491 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book focuses on bioelectrics, a new multidisciplinary field encompassing engineering and biology with applications to the medical, environmental, food, energy, and biotechnological fields. At present, 15 universities and institutes in Japan, the USA and the EU comprise the International Consortium of Bioelectrics, intended to advance this novel and important research field. This book will serve as an introductory resource for young scientists and also as a textbook for use by both undergraduate and graduate students – the world’s first such work solely devoted to bioelectrics.

Book Microfluidics and Lab on a Chip

Download or read book Microfluidics and Lab on a Chip written by Andreas Manz and published by Royal Society of Chemistry. This book was released on 2020-09-24 with total page 307 pages. Available in PDF, EPUB and Kindle. Book excerpt: Responding to the need for an affordable, easy-to-read textbook that introduces microfluidics to undergraduate and postgraduate students, this concise book will provide a broad overview of the important theoretical and practical aspects of microfluidics and lab-on-a-chip, as well as its applications.

Book Microfluidics

    Book Details:
  • Author : Yu Song
  • Publisher : John Wiley & Sons
  • Release : 2018-05-07
  • ISBN : 3527341064
  • Pages : 576 pages

Download or read book Microfluidics written by Yu Song and published by John Wiley & Sons. This book was released on 2018-05-07 with total page 576 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first book offering a global overview of fundamental microfluidics and the wide range of possible applications, for example, in chemistry, biology, and biomedical science. As such, it summarizes recent progress in microfluidics, including its origin and development, the theoretical fundamentals, and fabrication techniques for microfluidic devices. The book also comprehensively covers the fluid mechanics, physics and chemistry as well as applications in such different fields as detection and synthesis of inorganic and organic materials. A useful reference for non-specialists and a basic guideline for research scientists and technicians already active in this field or intending to work in microfluidics.

Book Insights in Nanobiotechnology 2022 2023  Novel Developments  Current Challenges  and Future Perspectives

Download or read book Insights in Nanobiotechnology 2022 2023 Novel Developments Current Challenges and Future Perspectives written by Gianni Ciofani and published by Frontiers Media SA. This book was released on 2023-12-12 with total page 138 pages. Available in PDF, EPUB and Kindle. Book excerpt: We are now entering the third decade of the 21st century, and, especially in the last years, the achievements made by scientists have been exceptional, leading to major advancements in the rapidly growing fields of bioengineering and biotechnology. This annual collection, which highlights article submissions from our Editorial Board members, looks to explore new insights, novel developments, current challenges, latest discoveries, recent advances, and future perspectives in the field of Nanobiotechnology. The Research Topic solicits brief, forward-looking contributions that describe the state of the art, outlining recent developments and major accomplishments that have been achieved and that need to occur to move the field forward. Authors are encouraged to identify the greatest challenges in the sub-disciplines, and how to address those challenges. The goal of this special edition Research Topic is to shed light on the progress made in the past decade and on its future challenges to provide a thorough overview of the nanobiotechnology field. This article collection will inspire, inform and provide direction and guidance to researchers. This collection is part of the 'Insights In' series which has been launched in each section of the journal. Other titles in the series include: Insights in Bioprocess Engineering 2022: Novel Developments, Current Challenges, and Future Perspective Insights in Biomaterials 2022: Novel Developments, Current Challenges, and Future Perspectives Insights In Biosafety & Biosecurity 2022: Novel Developments, Current Challenges, and Future Perspectives Insights in Biofabrication 2022: Novel Developments, Current Challenges, and Future Perspectives Insights in Tissue Engineering and Regenerative Medicine 2022 / 2023: Novel Developments, Current Challenges, and Future Perspectives Insights in Synthetic Biology 2022 / 2023: Novel Developments, Current Challenges, and Future Perspectives

Book Nanotechnology for Microfluidics

Download or read book Nanotechnology for Microfluidics written by Xingyu Jiang and published by John Wiley & Sons. This book was released on 2020-09-08 with total page 444 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book focuses on microfluidics with applications in nanotechnology. The first part summarizes the recent advances and achievements in the field of microfluidic technology, with emphasize on the the influence of nanotechnology. The second part introduces various applications of microfluidics in nanotechnology, such as drug delivery, tissue engineering and biomedical diagnosis.

Book Towards a New 3Rs Era in Experimental Research

Download or read book Towards a New 3Rs Era in Experimental Research written by Christopher R. Cederroth and published by Frontiers Media SA. This book was released on 2024-04-29 with total page 478 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the persisting need of animal experimentation for fundamental and applied research, the relevance and importance of the 3Rs Principle cannot be ignored. The 3Rs Principle was put forward over 50 years ago, providing an essential framework for more humane animal experimentation in research. In this half-century the research landscape within which this principle is applied has dramatically changed and evolved, with ever more emerging venues to explore for 3Rs advancement and implementation.