Abstract: In order to investigate the validity of relevant indicators in semen quality detection and guarantee the function of pig sperm, the fresh semen, pre-freezing semen（stored at 17 ℃ for two days） and thawed semen of five adult Rongchang boars were detected by automatic sperm analysis system. Artificial insemination tests were then carried out using frozen semen and sperm function was verified. Finally, plasma membrane integrity, mitochondrial normal rate and acrosomal integrity of fresh semen of the boar with poor insemination were analyzed by flow cytometry with normal semen before freezing as control. The results showed that the sperm of five adult Rongchang boars were normal in shape, good in density and motility, which met the requirements of relevant standards. No. 2 to No. 5 boars could breed normally, but sows bred with frozen semen of No. 1 boar were not pregnant. The plasma membrane integrity（account for 84.36%） and mitochondrial activity（account for 40.00%） of live sperm of No. 1 boar were higher than those of the control group（72.43% and 29.09%）, but the acrosomal integrity of live sperm of No. 1 boar was only 41.74%, which was much lower than that of the control group（91.83%）, indicating that the incomplete acrosomal sperm of No. 1 boar led to infertility. The results indicated that only detecting the shape, density and vitality of sperm before freezing could not guarantee the function of sperm. The integrity of plasma membrane, normal rate of mitochondria and integrity of acrosome of sperm should be controlled by flow cytometry to make up for the deficiency of traditional index detection, so as to ensure the healthy and sustainable development of pig industry.