4 6 diamidino 2phenylindole dapi staining h 1800

When genomic DNA is broken, the exposed 3′-OH ends can bind to fluorescein-labeled dUTP in the presence of terminal deoxynucleotidyl transferase (TdT), which can be detected using fluorescence microscopy. Tissue slices were deparaffinized with xylene, hydrated with a gradient of high percentage ethanol to distilled water, blocked with 20 μg/mL of a proteinase K solution for 30 min, processed with 0.5% Triton-X for 30 min, and then incubated with the rTDT reaction system (G3250, Promega, Shanghai, China) for 60 min at 37 °C in the dark. The reaction was then terminated using a 2× sodium citrate buffer for 15 min to terminate the reaction. Finally, 4′,6-diamidino-2phenylindole (DAPI) staining (H-1800, Vector Laboratories, USA) was used to define the nuclear area. The ratio of the number of apoptotic cells (green) and DAPI fluorescent spots (blue) was calculated by the Olympus IX81 microscope (Olympus, Japan).

When genomic DNA is broken, the exposed 3′-OH ends can bind to fluorescein-labeled dUTP in the presence of terminal deoxynucleotidyl transferase (TdT), which can be detected using fluorescence microscopy. Tissue slices were deparaffinized with xylene, hydrated with a gradient of high percentage ethanol to distilled water, blocked with 20 μg/mL of a proteinase K solution for 30 min, processed with 0.5% Triton-X for 30 min, and then incubated with the rTDT reaction system (G3250, Promega, Shanghai, China) for 60 min at 37 °C in the dark. The reaction was then terminated using a 2× sodium citrate buffer for 15 min to terminate the reaction. Finally, 4′,6-diamidino-2phenylindole (DAPI) staining (H-1800, Vector Laboratories, USA) was used to define the nuclear area. The ratio of the number of apoptotic cells (green) and DAPI fluorescent spots (blue) was calculated by the Olympus IX81 microscope (Olympus, Japan).