4.10. Immunofluorescence Staining on Testicular Sections

After deparaffinization of the testis sections, rehydrate them through a graded ethanol series. Then, perform antigen retrieval using a 0.1 mol/L citrate buffer solution at pH 6.0. The testicular sections were blocked with 3% BSA, 10% Donkey serum and 0.1% Triton X100 in PBS for 1 h at room temperature. Add the primary antibodies of QPRT (1:100, Proteintech) or VASA (1:100, Abcam) to the sections and incubate overnight at 4°C. After washing, add the corresponding fluorescent secondary antibodies (1:200, Alexa Fluor 555 and 488) and incubate at 37°C in the dark for 1 h. Following washing, mount the slides with an Antifade Mounting Medium with DAPI (Vector laboratories). The spermatocytes were observed and captured using immunofluorescence microscopy (Nikon, DS‐Qi2, Japan).