3.4. Hematoxylin and Eosin Staining

A 10% neutral buffered formalin solution was used to fix the testis for at least one week prior to HE staining. Approximately 3 mm thick testicular tissues were dehydrated, rendered transparent, and boiled in wax before being embedded in paraffin. The sections (3 μm) were dried for 48 h at 60 ° C and dewaxed in water before being dipped in hematoxylin (Sinopharm Chemical Reagent Beijing Co., Ltd., Beijing, China) for 15 min and washed in tap water for 5 min. Subsequently, the sections were dipped in eosin (Sinopharm Chemical Reagent Beijing Co., Ltd.) for 15 s, then washed again for 5 min. After dehydration of the alcohol gradient, the clearance with xylene and placement of a cover slip were performed. Observations were conducted using the LEICA DM6000 light microscope (Leica, Wetzlar, Germany).