The Multiplexing of Assays for the Measurement of Early Stages of Apoptosis by Polychromatic Flow Cytometry

Abstract

The detection of apoptosis has been a stalwart application for flow cytometric analysis for decades and this review of flow cytometric methods to detect early stages of apoptosis includes the use of the pivotal assay to detect early and late apoptosis, the Annexin V assay which when multiplexed with biologically functional fluorescent dyes to measure mitochondrial function and Reactive Oxygen Species (ROS) generation allows further identification of functionally different subsets within apoptotic populations. Here we show how this polychromatic approach can be used to demonstrate which subset of cells show changes in mitochondrial function and when ROS is generated in a time dependent manner. This polychromatic approach to flow cytometry leads to the identification of over ten sub-populations of cells during classic apoptosis or programmed cell death (PCD).