Introduction

Apoptosis, often called programmed cell death, is a carefully regulated process that is part of normal development and hemostasis. Because apoptosis is a dynamic event, and the time period during which cells exhibit apoptosis marker is variable and short, flow cytometry is an ideal technique for tracking cells through apoptosis.

Early Apoptosis Flow Cytometry Detection: There are two separate dyes that could identify apoptotic and non-apoptotic cells: Annexin V binds to phosphatidylserine on the external membrane of apoptotic cells, and 7-AAD permeates and stains DNA of late-stage apoptotic and dead cells.

Mid-Apoptosis Flow Cytometry Detection: During mid-apoptosis, we could measure Caspase activity using FLICA (fluorescent-labeled inhibitor of caspase) reagent, supplemented by a nuclear DNA stain 7-AAD, which evaluates membrane integrity and cell viability.

Late Apoptosis Flow Cytometry Detection: At the late apoptosis stage, we use the TUNEL assay to detect the apoptosis-induced DNA fragmentation through a quantitative fluorescence assay. Terminal deoxynucleotidyl transferase (TdT) catalyzes the incorporation of bromodeoxyuridine (BrdU) residues into the fragmented nuclear DNA at the 3’-hydroxyl ends. This assay distinguishes two populations: non-apoptotic cells (TUNEL-negative) and apoptotic cells (TUNEL-positive).

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