Flow Cytometry Protocols

Flow Cytometry Protocols

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Flow cytometry has evolved since the 1940s into a multidisciplinary field incorporating aspects of laser technology, fluid dynamics, electronics, optics, computer science, physics, chemistry, biology, and mathematics. Innovations in instrumentation, development of small lasers, discovery of new fluorochromes/fluorescent proteins, and implementation of novel methodologies have all contributed to the recent rapid expansion of flow cytometry applications. In this thoroughly revised and updated second edition of Flow Cytometry Protocols, time-proven as well as cutting-edge methods are clearly and comprehensively presented by leading experimentalists. In addition to being a valuable reference manual for experienced flow cytometrists, the editors expect this authoritative up-to-date collection to prove useful to investigators in all areas of the biological and biomedical sciences who are new to the subject. The introductory chapter provides an eloquent synopsis of the principles and diverse uses of flow cytometry, beginning with a historical perspective and ending with a view to the future. Chapters 2-22 contain step-by-step protocols of highly practical and state-of-the-art techniques. Detailed instructions and helpful tips on experimental design, as well as selection of reagents and data analysis tools, will allow researchers to readily carry out flow cytometric investigations ranging from traditional phenotypic characterizations to emerging genomics and proteomics applications. Complementing these instructive protocols is a chapter that provides a preview of the next generation of solid-state lasers, and one that describes a rapid means to validate containment of infectious aerosols generated during high-speed sorting (Chapters 23-24).show more

Product details

  • Paperback | 434 pages
  • 152.4 x 226.1 x 25.4mm | 861.84g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • Revised
  • 2nd ed. 2004
  • XIV, 434 p.
  • 1588292355
  • 9781588292353

Back cover copy

In this thoroughly revised and updated edition of a widely used practical guide to flow cytometry, leading experimentalists describe 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 fields 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. The protocols presented follow the successful Methods in Molecular Biology(TM) series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and highly practical, Flow Cytometry Protocols, Second Edition offers both basic and clinical scientists an up-to-date collection of cutting-edge protocols for solving the widest variety of novel flow cytometry problems.show more

Review quote

"This is a valuable addition to the literature and will be of benefit to research scientists and clinicians whether they be experienced or new to the methodology." - Cell Biochemistry and Function From Reviews of the First Edition... "...a well and precisely written collection of protocols which provide important information for both experienced and novice flow cytometry users." -FEBS Letters "...There are many people who would benefit from reading this book, from those currently using cytometry...to molecular biologists interested in looking at DNA or RNA in the context of cell phenotype in mixed cell populations and those involved in diagnostic testing..." -Molecular Biotechnologyshow more

Table of 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 ofFluorescence 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. Koupshow more