Multiparametric analysis of apoptosis by flow and image cytometry

Methods Mol Biol. 2004:263:141-60. doi: 10.1385/1-59259-773-4:141.

Abstract

Flow cytometric assays for apoptosis are now in widespread use. The multiparametric nature of flow cytometry allows multiple assays for several apoptotic characteristics to be combined in a single sample, providing a powerful tool for elucidating the complex progression of apoptotic death in a variety of cell types. This chapter describes one such assay, allowing simultaneous analysis of caspase activation, annexin V binding to "flipped" phosphatidylserine residues and membrane permeability to DNA binding dyes. This multidimensional 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.

MeSH terms

  • Animals
  • Annexin A5 / pharmacology
  • Apoptosis*
  • Caspases / metabolism
  • Coloring Agents / pharmacology
  • DNA / chemistry
  • Dactinomycin / analogs & derivatives*
  • Dactinomycin / pharmacology
  • Enzyme Activation
  • Flow Cytometry / methods*
  • Humans
  • Image Cytometry / methods*
  • Immunophenotyping / methods
  • Phosphatidylserines / chemistry
  • Propidium / pharmacology

Substances

  • Annexin A5
  • Coloring Agents
  • Phosphatidylserines
  • Dactinomycin
  • Propidium
  • 7-aminoactinomycin D
  • DNA
  • Caspases