2.3.4. Fluorescence assay (ethidium bromide/acridine orange staining) for apoptosis detection

Ethidium bromide and acridine orange (EB/AO) staining was used to determine the cyto-genotoxic nature of given compounds in testicular cells.²⁵ Briefly, the testicular cell suspension prepared from the treated testicular tissue was mixed with equal volume of EB/AO differential stain on a glass slide and covered with a coverslip. Observation and quantification of apoptotic cells were done with fluorescence microscope (Olympus, Japan) using filters of 500–525 nm. Cells appearing bright green were marked as normal healthy cells, while cells displaying orange-red fluorescence were marked as apoptotic.