Flow Cytometry Protocols

About this book

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 procedures.

Contents

1 Flow Cytometry: An Introduction Alice L. Givan 2 Multiparameter Flow Cytometry of Bacteria Howard M. Shapiro and Gerhard Nebe-von-Caron 3 Multiparameter Data Acquisition and Analysis of Leukocytes Carleton C. Stewart and Sigrid J. Stewart 4 Flow Cytometric Analysis of Kinase Signaling Cascades Omar D. Perez, Peter O. Krutzik, and Garry P. Nolan 5 Cytokine Flow Cytometry Holden T. Maecker 6 Use of Cell-Tracking Dyes to Determine the Proliferation Precursor Frequencies of Antigen-Specific T Cells Alice L. Givan, Jan L. Fisher, Mary G. Waugh, Nadege Bercovici, and Paul K. Wallace 7 Assessment of Lymphocyte-mediated Cytotoxicity Using Flow Cytometry Luzheng Liu, Beverly Z. Packard, Martin J. Brown, Akira Komoriya, and Mark B. Feinberg 8 Multiparametric Analysis of Apoptosis by Flow and Image Cytometry William G. Telford, Beverly Z. Packard, and Akira Komoriya 9 Detection and Enrichment of Hematopoietic Stem Cells by Side Population Phenotype Shannon S. Eaker, Teresa S. Hawley, Ali Ramezani, and Robert G. Hawley 10 Hematopoietic Stem Cell Characterization by Hoechst 33342 and Rhodamine 123 Staining Ivan Bertoncello and Brenda Williams 11 Phenotypic and Functional Analyses of Human CD34NEG Hematopoietic Precursors from Mobilized Peripheral Blood Douglas C. Dooley and Barbara K. Oppenlander 12 Multiparameter Flow Cytometry of Fluorescent Protein Reporters Teresa S. Hawley, Donald J. Herbert, Shannon S. Eaker, and Robert G. Hawley 13 The Analysis of Fluorescent Protein Expressing Cells by Flow Cytometry Steven C. Pruitt, Lawrence M. Mielnicki, and Carleton C. Stewart 14 Integrative Flow Cytometric and Microarray Approaches for Use in Transcriptional Profiling David W. Galbraith, Rangasamy Elumalai, and Fang Cheng Gong 15 Flow Cytometric Analysis of Fluorescence Resonance Energy Transfer: A Tool for High-throughput Screening of Molecular Interactions in Living Cells Francis Ka-Ming Chan and Kevin L. Holmes 16 Design of a FACS-based Mammalian Protein-Protein Interaction Trap (MAPPIT) Sam Lievens, Jose Van der Heyden, Els Vertenten, Jean Plum, Joel Vandekerckhove, and Jan Tavernier 17 Flow Cytometric Screening of Yeast Surface Display Libraries Michael Feldhaus and Robert Siegel 18 FRET-based HIV-1 Virion Fusion Assay Marielle Cavrois, Jason Neidleman, Martin Bigos, and Warner C. Greene 19 Cell Cycle Analysis of Asynchronous Populations Michael G. Ormerod 20 Solid Tumor DNA Content Analysis Adel K. El-Naggar and Philippe Vielh 21 Concurrent Flow Cytometric DNA and RNA Analysis Adel K. El-Naggar 22 Telomere Length Measurement by FISH and Flow Cytometry Veena Kapoor and William G. Telford 23 Small Lasers in Flow Cytometry William G. Telford 24 Viable Infectious Cell Sorting in a BSL-3 Facility Stephen P. Perfetto, David R. Ambrozak, Mario Roederer, and Richard A. Koup

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