Abstract
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.
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Acknowledgments
The authors wish to acknowledge Veena Kapoor and Nga Tu Voong of the National Cancer Institute for excellent technical assistance, and Dr. Z. Darzynkiewicz of the New York Medical College for helpful discussion. Bill Godfrey, Jolene Bradford, and Gayle Buller at Invitrogen Life Technologies (formerly Molecular Probes) provided valuable technical information regarding fluorescent probes. Parts of this work were supported by intramural research fund provided by the Center for Cancer Research, National Cancer Institute.
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Telford, W.G., Komoriya, A., Packard, B.Z., Bagwell, C.B. (2011). Multiparametric Analysis of Apoptosis by Flow Cytometry. In: Hawley, T., Hawley, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology, vol 699. Humana Press. https://doi.org/10.1007/978-1-61737-950-5_10
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DOI: https://doi.org/10.1007/978-1-61737-950-5_10
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