Cells that have lost plasma membrane integrity become permeable to external compounds, including dyes and enzymes, and are considered to be nonviable (5,7).
PI is a DNA dye. When the membrane on a nonviable cell becomes permeable, PI will enter the cell and bind to DNA in the nucleus. The binding of PI to DNA is then analyzed by the flow cytometer.
Perform a viability assessment for each specimen. Cell viability assessment by PI:
1. Lyse an aliquot of blood (tube 14). For tissue/fluid cell suspension without excess red blood cell contamination proceed to the next step.
2. Wash twice with PBS, decant.
3. Add 500 pL PI and incubate at room temperature in the dark for 10 min.
4. Document result on face sheet. Optimal viability is >80%.
5. Viability of <80% indicates poor viability.
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