Multiparametric analysis of apoptosis by flow cytometry

Methods Mol Biol. 2011:699:203-27. doi: 10.1007/978-1-61737-950-5_10.

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.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Annexin A5 / metabolism
  • Apoptosis* / drug effects
  • Apoptosis* / genetics
  • Caspases / metabolism
  • Cell Line, Tumor
  • Cycloheximide / pharmacology
  • DNA / metabolism
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods*
  • Fluorescent Dyes / metabolism
  • Immunophenotyping / methods*
  • Mice
  • Protein Synthesis Inhibitors / pharmacology
  • Staining and Labeling

Substances

  • Annexin A5
  • Fluorescent Dyes
  • Protein Synthesis Inhibitors
  • DNA
  • Cycloheximide
  • Caspases