Protocol

Flow Cytometry Protocols

Volume 699 of the series Methods in Molecular Biology pp 203-227

Date:

Multiparametric Analysis of Apoptosis by Flow Cytometry

  • William G. TelfordAffiliated withFACS Core Facilty, Experimental Transplantation and Immunology Branch, National Cancer Institute, National Institutes of Health
  • , Akira KomoriyaAffiliated withOncoImmunin, Inc.
  • , Beverly Z. PackardAffiliated withOncoImmunin, Inc.
  • , C. Bruce BagwellAffiliated withVerity Software House

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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.

Key words

Apoptosis Caspase Flow cytometry Annexin V 7-Aminoactinomycin D Propidium iodide Pacific Blue Hoechst 33258