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An easy quantitative cytotoxicity assay using a Coulter Counter
The Japanese Journal of Experimental Medicine
|April 1, 1981
Summary
This study introduces a new, objective cytotoxicity assay using an electronic particle counter. This rapid method offers a simpler and faster alternative to traditional dye exclusion for measuring cell death.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Cytotoxicity assays are crucial for evaluating immune responses and drug efficacy.
- Traditional dye exclusion methods can be subjective and time-consuming.
- Objective and rapid assays are needed for high-throughput screening.
Purpose of the Study:
- To develop a rapid and reproducible cytotoxicity assay using an electronic particle counter.
- To provide a more objective alternative to the dye exclusion method.
- To enable efficient monitoring of cytotoxic activity and titration of antibodies and complement.
Main Methods:
- Utilized an electronic particle counter (e.g., Coulter Counter) for cell counting.
- Employed anti-BAtheta antiserum and mouse thymocytes as target cells.
- Developed a procedure involving antibody/complement exposure, Pronase E digestion of dead cells, and Zap-Oglobin solubilization of viable cell nuclei for counting.
Main Results:
- Achieved comparable, yet more objective, results than the dye exclusion method.
- Demonstrated a significantly shorter assay time.
- Showed that contaminating erythrocytes do not interfere due to complete solubilization.
Conclusions:
- The described electronic particle counter-based assay is rapid, reproducible, and objective.
- This technique simplifies cytotoxicity measurements.
- It is highly applicable for high-volume analyses, including monitoring cytotoxic activity and titrating immune components.