Flow Cytometry Protocols (Methods in Molecular Biology, 1678)

This fourth edition volume expands on the previous editions by presenting readers with new and emerging methodologies in cytometry. The chapters in this book cover various topics such as quantifying surface and intracellular protein biomarkers, intracellular cytokine staining, apoptosis analysis, cell cycle analysis, tracking cell proliferation, and monitoring protein-protein interactions using FRET. Some developing methodologies that explored are mass cytometry, vesicle flow cytometry, time-resolved flow cytometry, and real-time label-free deformability cytometry. Written in the highly successful __Methods in Molecular Biology__ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and comprehensive, __Flow Cytometry Protocols, Fourth Edition researchers and scientists who are interested in continuing or expanding their knowledge of flow cytometry.__ Front Matter ....Pages i-xiv Flow Cytometry: The Glass Is Half Full (Howard M. Shapiro)....Pages 1-10 High-Dimensional Modeling for Cytometry: Building Rock Solid Models Using GemStoneTM and Verity Cen-se’TM High-Definition t-SNE Mapping (C. Bruce Bagwell)....Pages 11-36 Mass Cytometry Assays for Antigen-Specific T Cells Using CyTOF (Dongxia Lin, Holden T. Maecker)....Pages 37-47 RNA Flow Cytometry Using the Branched DNA Technique (Kah Teong Soh, Paul K. Wallace)....Pages 49-77 Analysis of Individual Extracellular Vesicles by Flow Cytometry (John P. Nolan, Erika Duggan)....Pages 79-92 Quantitative Fluorescence Measurements with Multicolor Flow Cytometry (Lili Wang, Adolfas K. Gaigalas, James Wood)....Pages 93-110 High Throughput Flow Cytometry for Cell Surface Profiling (Joshua Paterson, Laurie E. Ailles)....Pages 111-138 Multiparameter Conventional Flow Cytometry (Katherine M. McKinnon)....Pages 139-150 Multiparameter Intracellular Cytokine Staining (Patricia Lovelace, Holden T. Maecker)....Pages 151-166 Multiparametric Analysis of Apoptosis by Flow Cytometry (William G. Telford)....Pages 167-202 Multiparameter Cell Cycle Analysis (James W. Jacobberger, R. Michael Sramkoski, Tammy Stefan, Philip G. Woost)....Pages 203-247 Monitoring Cell Proliferation by Dye Dilution: Considerations for Probe Selection (Joseph D. Tario Jr., Alexis N. Conway, Katharine A. Muirhead, Paul K. Wallace)....Pages 249-299 Immunophenotypic Identification of Early Myeloerythroid Development (Cornelis J. H. Pronk, David Bryder)....Pages 301-319 Flow Cytometry Assays in Primary Immunodeficiency Diseases (Maurice. R. G. O’Gorman)....Pages 321-345 Real-Time Deformability Cytometry: Label-Free Functional Characterization of Cells (Maik Herbig, Martin Kräter, Katarzyna Plak, Paul Müller, Jochen Guck, Oliver Otto)....Pages 347-369 Nuclear Cytometry: Analysis of the Patterns of DNA Synthesis and Transcription Using Flow Cytometry, Confocal Microscopy, and RNA Sequencing (David W. Galbraith, Elwira Sliwinska, Partha Samadder)....Pages 371-392 Flow Cytometric FRET Analysis of Protein Interactions (László Ujlaky-Nagy, Péter Nagy, János Szöllősi, György Vereb)....Pages 393-419 Overview of Fluorescence Lifetime Measurements in Flow Cytometry (Jessica P. Houston, Zhihua Yang, Jesse Sambrano, Wenyan Li, Kapil Nichani, Giacomo Vacca)....Pages 421-446 Overview of Lasers for Flow Cytometry (William G. Telford)....Pages 447-479 Flow Cytometry: The Glass Is Half Empty (Howard M. Shapiro)....Pages 481-488 Back Matter ....Pages 489-492 This fourth edition volume expands on the previous editions by presenting readers with new and emerging methodologies in cytometry. The chapters in this book cover various topics such as quantifying surface and intracellular protein biomarkers, intracellular cytokine staining, apoptosis analysis, cell cycle analysis, tracking cell proliferation, and monitoring protein-protein interactions using FRET. Some developing methodologies that explored are mass cytometry, vesicle flow cytometry, time-resolved flow cytometry, and real-time label-free deformability cytometry. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Cutting-edge and comprehensive, Flow Cytometry Protocols, Fourth Edition is a valuable resource for researchers and scientists who are interested in continuing and/or expanding their knowledge of flow cytometry This thoroughly revised and updated edition of a widely used practical guide to flow cytometry describes in step-by-step detail an array of time proven and cutting-edge techniques much needed in today's advanced laboratories. These readily reproducible methods deploy emerging flow cytometry technologies in many new applications, especially in the field of stem cells, functional genomics and proteomics, and microbiology. Here, the aspiring investigator will find methods for the characterization of stem/progenitor cells by monitoring the efflux of fluorescent dyes and the elucidation of signal transduction pathways using phospho-specific antibodies. There are also techniques for monitoring gene transfer and expression using fluorescent protein technology, high throughput screening for discovery of novel protein interactions, phenotypic and functional characterization of T cell subsets and precursors, and microbial flow cytometry, to highlight but some of the many useful procedu.