Abstract
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.
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Acknowledgments
The author wishes to acknowledge Veena Kapoor and Nga Hawk of the National Cancer Institute for excellent technical assistance, and Dr. Zbigniew Darzynkiewicz of the New York Medical College for helpful discussion. Jolene Bradford and Gayle Buller at Thermo Fisher Life Technologies (formerly Molecular Probes) provided valuable technical information regarding fluorescent probes. Beverly Packard and Akira Komoriya also provided information regarding the PhiPhiLux probes, technical assistance and useful discussion. This work was supported by intramural research fund provided by the Center for Cancer Research, National Cancer Institute.
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Telford, W.G. (2018). Multiparametric Analysis of Apoptosis by Flow Cytometry. In: Hawley, T., Hawley, R. (eds) Flow Cytometry Protocols. Methods in Molecular Biology, vol 1678. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7346-0_10
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DOI: https://doi.org/10.1007/978-1-4939-7346-0_10
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