A sensitive method for accurate estimation of radiation induced apoptosis on peripheral blood mononuclear cells by acridine orange stain

A rapid and sensitive method for quantification of Apoptotic cells is presented. The value of the method is demonstrated on peripheral blood mononuclear cells, which constitutively undergo programmed cell death and which show an increase in the rate of apoptosis after exposure to the irradiation with 137 Cs γ -ray. After staining with Acridine orange post irradiation, three distinct cell populations (live, apoptosis, and necrosis) shown by analysis of fluorescence profiles. This Acridine orange stain method is directly compared with the stain method of Wright-Giemsa on duplicated slides and Apoptosis to quantify cell events following exposure to dose of in vitro γ -irradiation and post-irradiation incubation times. The apoptosis frequency of three healthy volunteers obtained with both stain methods was steep below 100cGy and flatter at high doses. A significantly higher number of radiation induced apoptotic cells were observed after stain with Acridine orange compared with Wright-Giemsa stain post irradiation. Our results demonstrated that the Acridine orange stain is superior methods with greater sensitivity and accuracy in comparison with the Wright-Giemsa stain tested. This method may be allows rapid measurement of a large number of cells for samples of radiation biological effect