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Cell-based Flow Cytometry Assay to Measure Cytotoxic Activity
Published on: December 17, 2013
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Measuring Cell Death by Propidium Iodide Uptake and Flow Cytometry.
Lisa C Crowley1, Adrian P Scott1, Brooke J Marfell1
1Apoptosis and Cytotoxicity Laboratory, Mater Research, Translational Research Institute, Woolloongabba, Brisbane, Queensland 4102, Australia;
Cold Spring Harbor Protocols
|July 3, 2016
Summary
Propidium iodide (PI) is a fluorescent stain used to identify dead cells. It enters cells with damaged membranes, allowing researchers to quantify cell death using flow cytometry.
Area of Science:
- Cell Biology
- Biotechnology
Background:
- Cell death detection is crucial in biological research.
- Distinguishing between live and dead cells requires reliable methods.
- Propidium iodide (PI) is a commonly used DNA-binding dye.
Purpose of the Study:
- To describe a protocol for using propidium iodide (PI) for cell death quantification.
- To highlight the utility of PI in flow cytometry for discriminating cell viability.
Main Methods:
- Propidium iodide (PI) staining protocol.
- Utilizing flow cytometry for cell analysis.
- Assessing plasma membrane integrity for cell discrimination.
Main Results:
- PI enters cells with compromised plasma membranes, indicating cell death.
- PI excludes from cells with intact plasma membranes, indicating viability.
- PI fluorescence is detectable with standard flow cytometry equipment.
Conclusions:
- Propidium iodide (PI) staining is an effective method for quantifying cell death.
- This protocol is adaptable for most research facilities and universities.
- PI facilitates reliable discrimination between live and dead cells based on membrane integrity.
