Multiparametric Analysis of Apoptosis by Flow Cytometry

Abstract

Flow cytometry remains the most widely used method for detecting and quantifying apoptosis and related forms of cell death in mammalian cells. The multiparametric nature of flow cytometry allows multiple apoptotic characteristics to be labeled and analyzed in a single sample, making it a powerful tool for analyzing the complex progression of apoptotic death. This chapter provides methods for combining assays for single apoptotic characteristics like caspase activation, annexin V binding, and cell membrane permeability into multiparametric assays that provide deeper insights into the cell death process. This approach to analyzing multiple apoptotic characteristics simultaneously yields far more information than single-parameter assays. While more informative than single-parameter assays, these multicolor methods can still be analyzed on relatively simple flow cytometers, making them widely accessible.

Keywords: 7-Aminoactinomycin D; Annexin V; Apoptosis; Caspase; Covalent viability probe; Flow cytometry; Fluorogenic caspase substrate; Hoechst dye; Necroptosis; Pacific Blue; Propidium iodide; Pyroptosis.