γH2AX staining and number of γH2AX foci in untreated (control) and irradiated blood cells.
Irradiation occurred with 2 Gy followed by 60 min post-incubation at room temperature before immobilization of PBMCs on the slides. (a) Representative samples. (A) PBMC after Ficoll density centrifugation. (B) Whole buffy coat after erythrolysis. (C) Blood smear from buffy coat without Ficoll gradient purification and erythrolysis. (D) Lymphocytes from a drop of blood from fingertip prepared according to the BDM. (b) Number of γH2AX foci in control and irradiated cells. Following irradiation with 2 Gy, cells were post-incubated for 60 min, transferred to slides, fixed and stained. (A) PBMCs after Ficoll density centrifugation. (B) Whole buffy coat after erythrolysis. (C) Whole buffy coat without erythrolysis. (D,E) Drop of blood from fingertip. (A–D) counting occurred by means of the Metafer quantification system (automatic scoring using the imageJ software); (E) counting of γH2AX foci occurred visually using LSM. Error bars indicate the intraexperimental variation in the PBMC population.