Dichlorodihydrofluorescein diacetate (DCFH-DA) assay completed at different time intervals on the human umbilical vein endothelial cells

DCFH-DA is a cell-permeable dye that is non-fluorescent and is hydrolysed intracellularly into its polar, but non-fluorescent form DCFH on the action of cellular esterase and thus is retained in the cell. The oxidation of DCFH by the action of intracellular reactive oxygen species (ROS) and other peroxides turns the molecule into its highly fluorescent form 2’,7’-dichlorofluorescein (DCF) that can be detected by various fluorescent methods.

The DCFH-DA assay was conducted on the endothelial cell line, EA.hy926 after exposure of varying concentrations of cadmium, chromium and lead alone and in combination at 3 different time intervals, namely 24 h, 48 h and 72 h.