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Enzyme Activity In Single Cells
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Book Synopsis Enzyme Activity in Single Cells by :
Download or read book Enzyme Activity in Single Cells written by and published by Academic Press. This book was released on 2019-11-12 with total page 326 pages. Available in PDF, EPUB and Kindle. Book excerpt: Enzyme Activity in Single Cells, Volume 628, the latest release in the Methods of Enzymology series, discusses groundbreaking cellular physiology research that is taking place in the biological sciences. Chapters in this new release cover Spatial and temporal resolution of caspase waves in single Xenopus eggs during apoptosis, Spatial and temporal organization of metabolic complexes in cells, Measuring cellular efflux and biomolecular delivery: synthetic approaches to imaging and engineering cells, Slide-based, single-cell enzyme assays, Single-cell assays using integrated continuous-flow microfluidics, High-throughput screening of single-cell lysates, Microfluidic capture of single cells for drug resistance assays, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology series Includes the latest information on Enzyme Activity in Single Cells
Book Synopsis Microfluidic Concentration-enhanced Single Cell Enzyme Activity Assay by : Aniruddh Sarkar
Download or read book Microfluidic Concentration-enhanced Single Cell Enzyme Activity Assay written by Aniruddh Sarkar and published by . This book was released on 2013 with total page 91 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cells sense stimuli, process information and respond using signaling networks regulated by enzymatic activity of various proteins. Aberrations in signaling are associated with diseases such as cancer. Most current methods lack the sensitivity to measure enzymatic activity in single cells and instead measure the average of large cell populations. Cellular heterogeneity, overlooked in these methods, is widespread and relevant. Microfabricated tools are uniquely suited to single cell analysis due to the match in size scale which enables high sensitivity, high throughput measurements. In this thesis we develop a microfluidic platform for the direct measurement of enzyme activities from selected single cells without disrupting their extracellular context. We develop modules to: enhance enzyme assay sensitivity by microfluidic confinement, interface microfluidic devices with selected single cells, enable multiplexing and then integrate these modules to perform single cell assays. We first investigate electrokinetic trapping of charged biomolecules in a nanofluidic concentrator for enhancing enzyme assay sensitivity by simultaneously accumulating enzyme and substrate into a reaction plug. Non-linear enhancement of reaction kinetics in this device is predicted by a mathematical model and experimentally verified. A linear enhancement mode is developed where only the enzyme is accumulated and is reacted with substrate later in an enclosed volume defined by integrated pneumatic valves or by micro-droplets formed using an integrated droplet generator. This device is then used to perform high-throughput measurement of secreted cellular proteases. We then develop a nicrofluidic probe for lysis and capture of the contents of selected single adherent cells from standard tissue culture platforms by creating a small lysis zone at its tip using hydrodynamic confinement. The single cell lysate is then divided and mixed with different substrates and confined in small chambers for fluorimetric assays. An integrated nanofluidic concentrator enables further concentration-enhancement. We demonstrate the ability to measure, from selected single cells, the activity of kinases: Akt, MAPKAPK2, PKA and a metabolic enzyme, GAPDH - separately or simultaneously. This assay platform can correlate single cell phenotype or extracellular context to intracellular biochemical state. We present preliminary explorations of the correlation of cell morphology or local cell population density to kinase activity.
Book Synopsis Molecular Biology of The Cell by : Bruce Alberts
Download or read book Molecular Biology of The Cell written by Bruce Alberts and published by . This book was released on 2002 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Single Cell Analysis of Biochemical Phenotypes by : Amanda L. Richer
Download or read book Single Cell Analysis of Biochemical Phenotypes written by Amanda L. Richer and published by . This book was released on 2018 with total page 221 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recent development of high throughput single cell methods has expanded our understanding of tissue heterogeneity, cell states, and developmental biology. Current single cell methods aim to understand cell function and phenotype by measuring a variety of cell features, like DNA sequence, mRNA abundance, chromatin accessibility, cell surface proteins, and histone modifications. However, cell phenotypes are regulated by a number of factors that escape the abundance measurements of current single cell methods. To directly measure cell phenotypes, I developed a method to measure enzyme activities in single cells. To this end, I designed DNA repair substrates that can be used to measure strand incision events catalyzed by endogenous DNA repair enzymes and called the method "Haircut". Haircut semi-quantitatively measures base excision repair and ribonucleotide excision repair and has been adapted to work simultaneously in a single cell mRNA sequencing experiment. Using Haircut, I measured mRNA expression and DNA repair activities in primary human immune cells and found differences in several DNA repair enzyme activities between immune cell types. Some of the repair activity measurements were supported by mRNA abundance measurements in those cell types. While other activities, especially those catalyzed by multimeric proteins, did not correlate with gene expression measurements. Additionally, I used Haircut to measure mRNA expression and DNA repair heterogeneity in immune cells from a preliminary cohort of individuals with trisomy 21 and found no differences in DNA repair between individuals with trisomy 21 and individuals with disomy 21. In summary, I developed a platform to measure enzymatic activities in single cells. I used Haircut to measure known and previously unknown differences in DNA repair across immune cell types. The platform I developed can be modified to measure other enzymatic activities and mRNA expression in thousands of single cells and can be expanded further to measure many enzymatic activities in millions of cells. Cell phenotypes are regulated by many complex mechanisms and are difficult to predict using DNA sequence or gene expression alone. Single cell analysis of biochemical phenotypes has the potential to bridge the gap between gene expression and cell function and provide direct functional readouts where other single cell methods cannot.
Book Synopsis Single Cell ‘Omics of Neuronal Cells by : Jonathan V. Sweedler
Download or read book Single Cell ‘Omics of Neuronal Cells written by Jonathan V. Sweedler and published by Springer Nature. This book was released on 2022-08-10 with total page 302 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume discusses the latest techniques used in the diverse fields of single cell ‘omics and covers topics such as quantifying the single cell transcriptome; isolation of cells in nanoliter volumes for single cell proteomics measurements by nano-LC-MS/MS; and single cell protein characterization by immunoblotting. A wide range of methodologies are highlighted, ranging from high-yield chemical amplification to mass spectrometry and nanotechnology for the analysis of the chemical constituents of cells. In the Neuromethods series style, chapters include both in-depth overviews, as well as detailed protocols that provide the key advice from specialists you need to get successful results in your laboratory. Cutting-edge and comprehensive, Single Cell ‘Omics of Neuronal Cells is a valuable resource for experienced and novice researchers interested in learning more about this field and its future developments.
Book Synopsis Enzymes of Molecular Biology by : Michael M. Burrell
Download or read book Enzymes of Molecular Biology written by Michael M. Burrell and published by Greenwood Publishing Group. This book was released on 1993-05-22 with total page 386 pages. Available in PDF, EPUB and Kindle. Book excerpt: Amasses key information on a broad range of enzymes commonly used as tools in molecular biology, condensing scattered sources for ease of reference, and making data accessible to nonenzymologists designing an experiment. After an overview of nucleases, 19 chapters give background on selected enzymes (nucleic acid- and protein-modifiers), discuss essential parameters for achieving optimized reactions, and provide exemplary practical procedures and protocols. Plastic comb binding. Annotation copyright by Book News, Inc., Portland, OR
Book Synopsis Biology for AP ® Courses by : Julianne Zedalis
Download or read book Biology for AP ® Courses written by Julianne Zedalis and published by . This book was released on 2017-10-16 with total page 1923 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biology for AP® courses covers the scope and sequence requirements of a typical two-semester Advanced Placement® biology course. The text provides comprehensive coverage of foundational research and core biology concepts through an evolutionary lens. Biology for AP® Courses was designed to meet and exceed the requirements of the College Board’s AP® Biology framework while allowing significant flexibility for instructors. Each section of the book includes an introduction based on the AP® curriculum and includes rich features that engage students in scientific practice and AP® test preparation; it also highlights careers and research opportunities in biological sciences.
Book Synopsis Essentials of Glycobiology by : Ajit Varki
Download or read book Essentials of Glycobiology written by Ajit Varki and published by CSHL Press. This book was released on 1999 with total page 694 pages. Available in PDF, EPUB and Kindle. Book excerpt: Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. "Essentials of Glycobiology" describes their biogenesis and function and offers a useful gateway to the understanding of glycans.
Book Synopsis Natural Biomarkers for Cellular Metabolism by : Vladimir V. Ghukasyan
Download or read book Natural Biomarkers for Cellular Metabolism written by Vladimir V. Ghukasyan and published by CRC Press. This book was released on 2014-09-26 with total page 404 pages. Available in PDF, EPUB and Kindle. Book excerpt: From the Lab to Clinical Settings-Advances in Quantitative, Noninvasive Optical DiagnosticsNoninvasive fluorescence imaging techniques, novel fluorescent labels, and natural biomarkers are revolutionizing our knowledge of cellular processes, signaling and metabolic pathways, the underlying mechanisms for health problems, and the identification of n
Book Synopsis Proteolytic Enzymes by : Erwin E. Sterchi
Download or read book Proteolytic Enzymes written by Erwin E. Sterchi and published by Springer Science & Business Media. This book was released on 2013-11-11 with total page 371 pages. Available in PDF, EPUB and Kindle. Book excerpt: Following an overview on proteolytic enzyme assays, this text covers procedures on how to investigate and study proteases. It describes the use of specific restriction proteases as well as inhibitors of proteases to prevent unwanted proteolysis.
Book Synopsis Biosensors for Single-Cell Analysis by : Jian Chen
Download or read book Biosensors for Single-Cell Analysis written by Jian Chen and published by Academic Press. This book was released on 2021-10-26 with total page 184 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biosensors for Single-Cell Analysis explores a wide range of biosensor technologies and their applications in single-cell characterization and analysis. Sections cover key biophysical and chemical single-cell properties that consider proteomic, metabolic, electrical, mechanical and optical properties. Each chapter features key definitions and case studies, providing detailed guidance for researchers who want to replicate covered solutions in their work. Tutorial sections, evaluations of the current state-of-the-field and future developments are also included. Microfluidic approaches to characterization, such as microfluidic impedance flow cytometry and microfluidic flow cytometry are considered alongside more conventional approaches, such as mass spectroscopy, fluorescent and mass flow cytometry. Additionally, key types of biosensors are covered, including atomic force microscopy, micropipette aspiration, optical tweezers, microfluidic hydrodynamic stretchers, microfluidic constriction channel and microfluidic optical stretchers. Includes chapters focused on key single-cell properties, such as proteomic, metabolic and mechanical characterization Features case studies that illustrate the application of biosensors for single-cell analysis Considers microfluidic approaches for each single-cell property discussed Explores future directions for single-cell analysis and biosensor technology
Download or read book Mechanisms of Catalysis written by and published by Academic Press. This book was released on 1991-01-28 with total page 471 pages. Available in PDF, EPUB and Kindle. Book excerpt: The remarkable expansion of information leading to a deeper understanding of enzymes on the molecular level necessitated the development of this volume which not only introduces new topics to The Enzymes series but presents new information on some covered in Volume I and II of this edition.
Book Synopsis Concepts of Biology by : Samantha Fowler
Download or read book Concepts of Biology written by Samantha Fowler and published by . This book was released on 2017-12-30 with total page 624 pages. Available in PDF, EPUB and Kindle. Book excerpt: Concepts of Biology is designed for the introductory biology course for nonmajors taught at most two- and four-year colleges. The scope, sequence, and level of the program are designed to match typical course syllabi in the market. Concepts of Biology includes interesting applications, features a rich art program, and conveys the major themes of biology. The images in this textbook are grayscale.
Book Synopsis Modern Fluorescence Spectroscopy by : E. L. Wehry
Download or read book Modern Fluorescence Spectroscopy written by E. L. Wehry and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the appearance of the first two volumes of Modern Fluorescence Spectroscopy in 1976, important advances continue to be made in both the techniques and applications of molecular luminescence. In terms of "hardware," it is only recently that the application of laser excitation to molecular fluorometry has become feasible under conditions that are analy tically realistic. The improvements that can be effected in sensitivity, analy tical selectivity, and ability to handle "difficult" samples by laser fluorometry have only begun to be exploited. Likewise, time-resolved fluorometry has received widespread use in fundamental studies (a sizable number of which deal with biological systems), but has as of yet received relatively little analytical utilization. The use of electronic array detectors offers the promise of obtaining luminescence spectra more rapidly, and perhaps ultimately with greater sensitivity, than is possible by the use of scanning instruments equipped with conventional detectors. The increasing capabilities of microcomputers and the increasing sophistication of "smart" spectroscopic instrumentation signify that much more efficient acquisition and use can now be achieved of the information contained in the "excitation-emission matrix" inherent in the luminescence phenomenon.
Book Synopsis Trends in Enzyme Histochemistry and Cytochemistry by : David Evered
Download or read book Trends in Enzyme Histochemistry and Cytochemistry written by David Evered and published by John Wiley & Sons. This book was released on 2009-09-16 with total page 192 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Novartis Foundation Series is a popular collection of the proceedings from Novartis Foundation Symposia, in which groups of leading scientists from a range of topics across biology, chemistry and medicine assembled to present papers and discuss results. The Novartis Foundation, originally known as the Ciba Foundation, is well known to scientists and clinicians around the world.
Book Synopsis Cells: Molecules and Mechanisms by : Eric Wong
Download or read book Cells: Molecules and Mechanisms written by Eric Wong and published by Axolotl Academic Publishing. This book was released on 2009 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: "Yet another cell and molecular biology book? At the very least, you would think that if I was going to write a textbook, I should write one in an area that really needs one instead of a subject that already has multiple excellent and definitive books. So, why write this book, then? First, it's a course that I have enjoyed teaching for many years, so I am very familiar with what a student really needs to take away from this class within the time constraints of a semester. Second, because it is a course that many students take, there is a greater opportunity to make an impact on more students' pocketbooks than if I were to start off writing a book for a highly specialized upper- level course. And finally, it was fun to research and write, and can be revised easily for inclusion as part of our next textbook, High School Biology."--Open Textbook Library.
Download or read book Chromatography written by E. Heftmann and published by Elsevier. This book was released on 2004-04-16 with total page 585 pages. Available in PDF, EPUB and Kindle. Book excerpt: Chromatography has emerged as the most important and versatile analytical method. The book is not only an updated version of Heftmann's classical text, but it covers areas of future importance, such as microfluidics and computer resources. Under his experienced guidance, authorities in each field have contributed their practical experience to an integrated treatment of modern micro analysis. In Part A the theoretical basis of individual separation methods is explained and the technical aspects are illustrated. It includes the theory of gas and liquid chromatography as well as specific chromatographic techniques, such as size-exclusion, planar, ion, and affinity chromatography as well as various electrokinetic separation techniques. Microfluidics are covered for the first time and useful sources of analytical instruments are listed and evaluated. 1. Each chapter written by an authority2. Thorough treatment of the theoretical basis of separation methods3. Practical guide for performing analyses
