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Published in

Humana Press, Methods in Molecular Biology, p. 11-20, 2014

DOI: 10.1007/978-1-4939-1661-0_2

Apoptosis and Cancer, p. 23-33

DOI: 10.1007/978-1-59745-339-4_3

Apoptosis and Cancer, p. 23-34

DOI: 10.1385/1-59745-339-0:23

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Flow Cytometry Enumeration of Apoptotic Cancer Cells by Apoptotic Rate

This paper is available in a repository.
This paper is available in a repository.

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Abstract

Most authors currently quantify the frequency of apoptotic cells in a given phenotypically defined population after calculating the apoptotic index (AI), that is, the percentage of apoptotic cells displaying a specific lineage antigen (LAg) within a population of cells that remain unfragmented and retain the expression of the LAg. However, this approach has two major limitations. First, apoptotic cells fragment into apoptotic bodies that later disintegrate. Second, 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 will describe a flow cytometry method to calculate the apoptotic rate (AR) that takes into account both cell fragmentation and loss of LAg 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.