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Trypan blue exclusion test of cell viability
1National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
The dye exclusion test identifies live cells in a suspension. Viable cells exclude dyes like trypan blue, appearing clear, while dead cells absorb the dye, appearing blue.
Area of Science:
- Cell biology
- Biotechnology
Background:
- Cell viability assessment is crucial in biological research.
- Dye exclusion assays offer a straightforward method for this determination.
Purpose of the Study:
- To outline the principle and application of the dye exclusion test.
- To describe a protocol for distinguishing viable from nonviable cells.
Main Methods:
- Mixing a cell suspension with a vital stain (e.g., trypan blue).
- Microscopic examination to differentiate cells based on dye uptake.
- Viable cells exclude dye, showing clear cytoplasm; nonviable cells absorb dye, showing stained cytoplasm.
Main Results:
- Successful differentiation between live and dead cells based on cytoplasmic staining.
- Quantification of viable cells within a population.
Conclusions:
- The dye exclusion test is a reliable method for assessing cell viability.
- This technique is fundamental for cell-based assays and experiments.
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