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LIVE/DEAD Viability/Cytotoxicity Assay for Animal Cells Using the SpectraMax Gemini XS Fluorescence Microplate Reader (MaxLine Application Note #43)

fluorescent signal approximately equal to 2.5 x 103 dead cells/well. This may be a result of fluorescence interference or a small (10%) number of contaminating dead cells within the live cell population. These two figures demonstrate that there is a significant difference in fluorescent signal between live and dead cells that easily differentiates the two cell populations when they are stained with a mixure of the dyes.



CONCLUSION
The LIVE/DEAD Viability/Cytoxicity assay is a simple and fast method for detection of live and dead cells. Using the optimized wavelength settings, the SPECTRAmax GEMINI XS fluorescence microplate reader can distinguish between live and dead cells using this assay. The data suggests that there is little interference from dead cells when live cells are analyzed, however, there may be some interference from live cells when dead cells are analyzed. This assay should be especially suitable for growth inhibition and cell death assays that emphasize determination of a relative decrease in live cell number using cal AM dye. A standard curve for both live and dead cells is easily generated using the SOFTmax PRO software and its integrated formulas.


REFERENCES
1. Wagner, M. M., Paul, D. C., Shih, C., Jordan, M. A., Wilson, L., and Williams, D. C. In vitro pharmacology of cryptophycin 52 (LY355703) in human tumor cell lines. Cancer Chemother. Pharmacol. 43: 115-125 (1999).

2. Adler, M., Shafer, H., Hamilton, T., and Petrali, J. P. Cytotoxic actions of the heavy metal chelator TPEN on NG108-15 neuroblastoma-glioma cells. Neurotoxicity 20: 571-582 (1999).

3. Papadopoulos, N. G., Dedoussis, G. V., Spanakos, G., Gritzapis, A. D., Baxevanis, C. N., and Papamichail, M. An improved fluorescence assay for the determination of lymphocyte-me
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