A novel and rapid apoptosis assay based on thiol redox status

We present here a novel probe, VitaBright-48, for the evaluation of the cellular level of reduced thiols. Using different cell lines and apoptogenic agents we show that a decrease in the level of reduced thiols correlates with well-known apoptotic markers such as phosphatidylserine translocation and caspase activity. The cell population to be investigated is added to the nonfluorescent stain VitaBright-48, which immediately permeates the cell membrane and reacts with intracellular thiols, forming a fluorescent compound. Quantification of the cell fluorescence directly after staining (without washing) can then be used to determine the population's cellular thiol level at the single cell level. Based on the results presented here, we suggest that measurement of changes in the level of free thiols should be added to the list of phenotypes which may be investigated in order to detect apoptosis. © 2012 International Society for Advancement of Cytometry