Download or read book Development of a Novel Microfluidic Platform to Study T Cell Signaling written by Shannon L. Faley and published by . This book was released on 2007 with total page 109 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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.
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
Download or read book Development of Microfluidic Cell Culture Platforms for Investigating Cellular Phenomena written by Bryson McKinley Brewer and published by . This book was released on 2015 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Studying Cell Metabolism and Cell Interactions Using Microfluidic Devices Coupled with Mass Spectrometry written by Huibin Wei and published by Springer Science & Business Media. This book was released on 2012-10-17 with total page 117 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis describes a new approach for cell analysis by the rapid developing microfluidic technology. The nominee has made great contributions to develop a new analysis platform which combined microfluidic devices with mass spectrometry to determine the trace compounds secreted by cells. Based on this analysis platform, she studied the specific cell secreting behaviors under controlled microenvironment, of which the secretion compounds were qualified and semi-quantified by mass spectrometry. A novel cell sorting device integrated homogenous porous PDMS membrane was invented to classify cells from real samples based on the size difference. The nominee further studied the signal transmission between different cells, and the signal chemicals were qualitative and quantitative monitored by the analysis platform. This indicates the potential significant application of the new cell analysis platform in medicine screening and early diagnosis.
Download or read book Microfluidic Methods for Molecular Biology written by Chang Lu and published by Springer. This book was released on 2016-05-14 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the state-of-the-art research on molecular biology assays and molecular techniques enabled or enhanced by microfluidic platforms. Topics covered include microfluidic methods for cellular separations and single cell studies, droplet-based approaches to study protein expression and forensics, and microfluidic in situ hybridization for RNA analysis. Key molecular biology studies using model organisms are reviewed in detail. This is an ideal book for students and researchers in the microfluidics and molecular biology fields as well as engineers working in the biotechnology industry. This book also: Reviews exhaustively the latest techniques for single-cell genetic, epigenetic, metabolomic, and proteomic analysis Illustrates microfluidic approaches for inverse metabolic engineering, as well as analysis of circulating exosomes Broadens readers’ understanding of microfluidics convection-based PCR technology, microfluidic RNA-seq, and microfluidics for robust mobile diagnostics
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.
Download or read book Development of Novel Microfluidic Platforms for Neural Stem Cell Research written by Bonggeun Chung and published by . This book was released on 2007 with total page 236 pages. Available in PDF, EPUB and Kindle. Book excerpt: To make novel microfluidic devices integrated with multiple functions, we have also developed a microfluidic multi-injector (MMI) that can generate temporal and spatial concentration gradients. MMI consists of fluidic channels and control channels with pneumatically actuated on-chip barrier valves. Repetitive actuations of on-chip valves control pulsatile release of solution that establishes microscopic chemical gradients. The development of novel gradient-generating microfluidic platforms will help in advancing our understanding of brain development and provide a versatile tool with basic and applied studies in stem cell biology.
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.
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.
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.
Download or read book Development of Multi purpose Microfluidic Platform for Single cell and Pairwise Cell Interaction Assay written by Young-Ji Kim and published by . This book was released on 2009 with total page 95 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Microfluidic Organ on a Chip Revolutionary Platforms for Disease Comprehension and Treatment written by Shi-Cong Tao and published by Frontiers Media SA. This book was released on 2022-01-11 with total page 176 pages. Available in PDF, EPUB and Kindle. Book excerpt: Existing culture systems have a limited ability to reproduce the complicated and dynamic microenvironment of a functioning organ. To solve the issues of conventional culture techniques, multidisciplinary researchers, involving medical doctors, stem cell and developmental biology experts, engineers and physical scientists, have emerged to innovate methods and devices. A microfluidic organ-on-a-chip (μOOC) is a cell culture device, based on microfluidic technology, which contains continuously perfused chambers with cells to simulate organ-level physiology/pathology. The μOOC is not to build a whole living organ, but rather to synthesize minimal functional units that recapitulate organ-/tissue-level functions. The μOOC can be applied to study not only the convention stimulation on cells by molecular/drugs, but also physical forces (fluid shear stress, cyclic strain and mechanical compression), organ-specific cell-cell intercommunication, and organ-organ coupling responses. There is an emerging need for innovative approaches for the production, control, analysis, and utilization of the μOOC, and even the multiple interconnected μOOC (Human-on-a-Chip). Although the μOOC has attracted much attention and is continuous being studied, there are still many difficult problems to be solved. Some of the most mentioned challenges include microenvironmental (biochemical, biophysical, biomechanical, nutrient, etc.) control, modeling tissue–tissue and multiorgan interactions, and reducing variability (automated control, high-throughput manipulation/analysis, integration of biosensing and etc.).
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.
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.
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.
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.
