Measuring Cell Death by Propidium Iodide Uptake and Flow Cytometry
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Updated: Mar 18, 2026

Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
Published on: December 17, 2013
Lisa C Crowley1, Adrian P Scott1, Brooke J Marfell1
1Apoptosis and Cytotoxicity Laboratory, Mater Research, Translational Research Institute, Woolloongabba, Brisbane, Queensland 4102, Australia;
Abstract:
Propidium iodide (PI) is a small fluorescent molecule that binds to DNA but cannot passively traverse into cells that possess an intact plasma membrane. PI uptake versus exclusion can be used to discriminate dead cells, in which plasma membranes become permeable regardless of the mechanism of death, from live cells with intact membranes. PI is excited by wavelengths between 400 and 600 nm and emits light between 600 and 700 nm, and is therefore compatible with lasers and photodetectors commonly available in flow cytometers. This protocol for PI staining can be used to quantitate cell death in most modern research facilities and universities.

