Method of differential staining of sister chromatids for the study of the effects of gamma rays on the frequency of sister chromatid exchange in human lymphocytes]

Human lymphocytes were incubated during two cycles of replication in the presence of 5-bromodeoxyuridine, fixed after a 96 hours cultivation, stained with fluorescenct compound "Hoechst 33258", illuminated with sunlight and repeatedly stained with azureosine. After such a treatment, the two chromatids of metaphase chromosomes are stained with different intensity revealing numerous sister chromatid exchanges (SCE) which could be exactly recorded. In spite of the use if tge standard technique, the frequency of SCE was different in two donors. Irradiation after a 47 hours incubation (mainly G2 stage of the first cycle) increased the frequency of SCE, whereas the irradiation 2 hours before fixation (G2 stage of the second cycle) decreased it. The change of the frequence of SCE produced by irradiation was not proportional to the chromosome length.