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51Cr-release assay adapted to a 96-well format sample reading
G G Hillman1, N Roessler, R S Fulbright
1Department of Urology, Wayne State University School of Medicine, Detroit, MI 48201.
Biotechniques
|October 1, 1993
Summary
The Matrix 96, a direct beta counter, accurately monitors cell-mediated lympholysis assays (CML). This method efficiently measures chromium-51 release in 96-well plates, correlating well with traditional gamma counters.
Area of Science:
- Immunology
- Biotechnology
- Analytical Chemistry
Background:
- Cell-mediated lympholysis (CML) assays traditionally use gamma counters for chromium-51 (51Cr) detection.
- This method requires individual tube readings, limiting throughput.
- A need exists for more efficient CML assay monitoring.
Purpose of the Study:
- To evaluate the Matrix 96, a 96-well direct beta counter, for monitoring CML assays.
- To compare the accuracy and efficiency of the Matrix 96 with conventional gamma counters for 51Cr detection.
Main Methods:
- Cell-mediated lympholysis assays were performed in 96-well microplates.
- Supernatants containing released 51Cr were spotted onto a 96-well metal plate and dried.
- Samples were read using the Matrix 96 direct beta counter and compared to a standard gamma counter.
Main Results:
- A strong correlation was found between 51Cr counts obtained from the Matrix 96 and the gamma counter.
- The Matrix 96 accurately detected beta particles from 51Cr, comparable to gamma ray detection.
- Both methods confirmed low cytotoxicity by unstimulated lymphocytes and high activity by lymphokine-activated killer (LAK) cells.
Conclusions:
- The Matrix 96 is a viable and accurate alternative for monitoring CML assays.
- This direct beta counter offers increased efficiency for high-throughput analysis of 51Cr release.
- The findings support the use of Matrix 96 in immunological studies involving cytotoxicity assays.