Flow Cytometry Protocols

William G. Telford

Flow cytometry is the most widely used method for detecting and quantifying apoptosis in mammalian cells. 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 single-apoptosis assays such as fluorogenic caspase substrates, annexin V binding, DNA dye exclusion, and covalent viability probes into informative multiparametric assays. This multiparametric approach to analyzing apoptosis provides much more information than single-parameter assays that provide only a percentage apoptotic result, given that multiple early, intermediate, and late apoptotic stages can be observed and quantified simultaneously. While much more informative than single-color assays, these multicolor methods can still be analyzed on relatively simple flow cytometers, making them accessible to many laboratories.