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In situ screening assay for cell viability using a dimeric cyanine nucleic acid stain
B Becker1, J Clapper, K R Harkins
1Department of Biochemistry and Biophysics, Iowa State University, Ames 50011.
Analytical Biochemistry
|August 15, 1994
Summary
This study introduces a new cell viability assay using YOYO-1 dye for accurate measurement in microplates. The assay offers a sensitive and rapid method for assessing cell health in various cell types.
Area of Science:
- Cell Biology
- Biotechnology
- Assay Development
Background:
- Accurate cell viability assessment is crucial for biological research and drug discovery.
- Existing methods like flow cytometry and metabolic assays have limitations in speed, sensitivity, or inclusivity of all cell types.
Purpose of the Study:
- To develop and validate a rapid, sensitive, and in situ microplate assay for determining cell viability.
- To utilize a novel dye exclusion method for improved accuracy and comprehensive cell analysis.
Main Methods:
- Developed a dye exclusion assay using the plasma membrane-impermeable cyanine dye YOYO-1.
- Performed assay in situ in 96-well microtiter plates, measuring fluorescence before and after detergent treatment.
- Calculated cell viability based on the ratio of fluorescence from nonviable and total cells.
Main Results:
- The YOYO-1 assay demonstrated high sensitivity and rapidity for both adherent and nonadherent cells.
- Results closely correlated with flow cytometry dye exclusion studies.
- The assay effectively included disintegrated cells, aggregates, and adherent cells in measurements, unlike some other methods.
Conclusions:
- The YOYO-1 microplate assay provides a robust and comprehensive method for in situ cell viability determination.
- This assay is a valuable tool for biological research, offering advantages over traditional methods.
- The in situ nature ensures all cell populations are accounted for, enhancing data reliability.