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.