Evaluation of cell viability by double-staining fluorescence assay.

Various methods are available for the assessment of cell viability. Recently, interest has centered on methods using fluorescent dyes. In this work, we used a double-staining assay, involving the use of rhodamin 123, which stains the mitochondria of viable cells, and propidium iodide, which stains the nuclei of dead cells, to investigate their use in assessing the viability of cells. We added adriamycin to NIH 3T3 cells and double-stained the cells that adhered to the dish and those that were suspended in the culture solution, and observed the results over time. We found that nearly all the adherent cells were stained with rhodamin 123 alone. However, the suspended cells in the control group accounted for most of the double-stained cells, and when adriamycin was added, most were stained with PI alone.