Yu Lin, Cheng Qing-Yuan, Jia Ye-Lin, Zheng Yan, Yang Ting-Ting, Wu Ying-Bi, Li Fu-Ping
Department of Andrology/Sichuan Human Sperm Bank, West China Second University Hospital, Sichuan University, Chengdu 610000, China.
Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Chengdu 610000, China.
Asian J Androl. 2025 Jan 1;27(1):59-64. doi: 10.4103/aja202474. Epub 2024 Sep 3.
According to the World Health Organization (WHO) manual, sperm concentration should be measured using an improved Neubauer hemocytometer, while sperm motility should be measured by manual assessment. However, in China, thousands of laboratories do not use the improved Neubauer hemocytometer or method; instead, the Makler counting chamber is one of the most widely used chambers. To study sources of error that could impact the measurement of the apparent concentration and motility of sperm using the Makler counting chamber and to verify its accuracy for clinical application, 67 semen samples from patients attending the Department of Andrology, West China Second University Hospital, Sichuan University (Chengdu, China) between 13 September 2023 and 27 September 2023, were included. Compared with applying the cover glass immediately, delaying the application of the cover glass for 5 s, 10 s, and 30 s resulted in average increases in the sperm concentration of 30.3%, 74.1%, and 107.5%, respectively (all P < 0.0001) and in the progressive motility (PR) of 17.7%, 30.8%, and 39.6%, respectively (all P < 0.0001). However, when the semen specimens were fixed with formaldehyde, a delay in the application of the cover glass for 5 s, 10 s, and 30 s resulted in an average increase in the sperm concentration of 6.7%, 10.8%, and 14.6%, respectively, compared with immediate application of the cover glass. The accumulation of motile sperm due to delays in the application of the cover glass is a significant source of error with the Makler counting chamber and should be avoided.
根据世界卫生组织（WHO）手册，精子浓度应使用改良的Neubauer血细胞计数板进行测量，而精子活力应通过人工评估来测量。然而，在中国，数以千计的实验室并未使用改良的Neubauer血细胞计数板或方法；相反，Makler计数室是使用最广泛的计数室之一。为了研究可能影响使用Makler计数室测量精子表观浓度和活力的误差来源，并验证其在临床应用中的准确性，纳入了2023年9月13日至2023年9月27日期间在四川大学华西第二医院男科就诊的67例患者的精液样本。与立即盖上盖玻片相比，延迟5秒、10秒和30秒盖上盖玻片，精子浓度平均分别增加30.3%、74.1%和107.5%（所有P<0.0001），进行性活力（PR）平均分别增加17.7%、30.8%和39.6%（所有P<0.0001）。然而，当精液标本用甲醛固定后，与立即盖上盖玻片相比，延迟5秒、10秒和30秒盖上盖玻片，精子浓度平均分别增加6.7%、10.8%和14.6%。由于延迟盖上盖玻片导致活动精子的积累是Makler计数室误差的一个重要来源，应予以避免。
