Diaz David, Prieto Alfredo, Reyes Eduardo, Barcenilla Hugo, Monserrat Jorge, Alvarez-Mon Melchor
CNB-CSIC R&D Associated Unit, Department of Medicine, University of Alcalá, Carretera Madrid-Barcelona, Km 33.600, 28871, Alcalá de Henares, Madrid, Spain,
Methods Mol Biol. 2015;1219:11-20. doi: 10.1007/978-1-4939-1661-0_2.
Most authors currently quantify the frequency of apoptotic cells in a given phenotypically defined population after calculating the apoptotic index (AI), i.e., the percentage of apoptotic cells displaying a specific linage antigen (LAg) within a population of cells that remain unfragmented and retain the expression of the LAg. However, this approach has two major limitations. Firstly, apoptotic cells fragment into apoptotic bodies that later disintegrate. Secondly, apoptotic cells frequently lose, partially or even completely, the cell surface expression of the LAg used for the identification of specific cell subsets. This chapter describes a flow cytometry method to calculate the apoptotic rate (AR) that takes into account both cell fragmentation and loss of lineage antigen expression on measurement of apoptosis using flow cytometry ratiometric cell enumeration that emerges as a more accurate method of measurement of the occurrence of apoptosis in normal and tumoral cell cultures.
