Sperm Analysis from Epididymis in Mice

The caudal epididymis was removed from the anesthetic adult mice treated with or without LY411575 for 35 days and cut by eye scissors into small bulks. Next, these tissues were submerged in 500 μl of Tyrode’s salt solution (T2397, Sigma, U.S.A.) at 5% CO₂, 37°C incubator for 15 min. The sperms swam out into the salt solution after incubation, and were obtained by centrifugation at 600 g for 5 min at room temperature. Next, the sediments were re-suspended with 1 ml of PBS. Two aliquots of suspension (each 10 μl) were taken out to perform with computer-assisted sperm analysis software (CASA software) as previously described and Giemsa staining (G1020, Solarbio, China) was performed according to the production manual [16 (link)]. The length of the sperm midpiece was measured using ImageJ software.