Multiparametric analysis of apoptosis by flow cytometry.
Flow cytometry is the most widely used technology for analyzing apoptosis. The multiparametric nature of flow cytometry allows several apoptotic characteristics to be combined in a single sample, making it a powerful tool for analyzing the complex progression of apoptotic death. This chapter provides guidelines for combining caspase detection, annexin V binding, DNA dye exclusion, and other single apoptotic assays into multiparametric assays.This approach to analyzing apoptosis provides far more information than single parameter assays that provide only an ambiguous "percent apoptotic" result, given that multiple early, intermediate and late apoptotic stages can be visualized simultaneously. This multiparametric approach is also amenable to a variety of flow cytometric instrumentation, both old and new.
FACS Core Facilty, Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
William G
WG
Telford
Akira
A
Komoriya
Beverly Z
BZ
Packard
C Bruce
CB
Bagwell
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D016130:Immunophenotyping / Q000379:methods*; D051379:Mice; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013194:Staining and Labeling
D017304:Annexin A5; D005456:Fluorescent Dyes; D011500:Protein Synthesis Inhibitors; D004247:DNA; D003513:Cycloheximide; D020169:Caspases