A new method for evaluating the quality of single sperm by detecting reactive oxygen species  

作　　者：Hao Hu Bei Yan Wei Zhang Wenbo Yan Liquan Liu Xiaofeng Tang Kejun Dong Longjie Li Xianjin Xiao Chengliang Xiong 

机构地区：[1]Institute of Reproductive Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China [2]Sperm Bank,General Hospital of Ningxia Medical University,Yinchuan 750004,China [3]Department of Obstetrics and Gynecology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China [4]School of Life Science and Technology,Wuhan Polytechnic University,Wuhan 430023,China [5]Reproductive Medicine Center,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China [6]Hubei Province Human Sperm Bank,Wuhan 430030,China

出　　处：《Chinese Chemical Letters》2023年第2期306-310,共5页中国化学快报（英文版）

基　　金：supported by the National Natural Science Foundation of China (Nos. 21705053 and 81871732);the Open Foundation of NHC Key Laboratory of Birth Defects Prevention (No. ZD202104);the Open Foundation of Key Laboratory of Birth Regulation and Control Technology of National Health Commission of China, Maternal Child Health Care Hospital of Shandong Province。

摘　　要：Sperm damage caused by reactive oxygen species(ROS) is one of the main causes of male infertility.Therefore, the level of ROS in sperm is an important indicator for the diagnosis and prognosis of male infertility. Herein, we constructed a single sperm ROS detection method(SSRDM) with an optical microprobe fabricated via focused ion beam process. The micro-probe is used to separately excite fluorescence in the sperm and the area around the sperm after ROS staining, and the difference in fluorescence values can reflect the level of ROS in the sperm. We collected 102 semen samples and 72 of them were divided into asthenozoospermia and non-asthenozoospermia groups. SSRDM and flow cytometry were used to detect the ROS levels of the two groups. The results of SSRDM showed that the ROS levels of asthenozoospermia group were higher than that of non-asthenozoospermia group(P = 0.002), while the results of flow cytometry indicated no difference(P = 0.152). In terms of ROS levels, compared with flow cytometry, SSRDM has a stronger ability to distinguish between those two groups, providing a reliable basis for assessment of sperm quality. Another 30 semen samples were used to investigate temperature and temporal variability of SSRDM to ensure the stability and accuracy of this method. Overall, we have developed a method that can quantitatively detect fluorescent substances in sperm at the single-cell level supplying evidence for diagnosis and prognosis of male infertility.

关 键 词：Reactive oxygen species Single sperm Male infertility MICROPROBE Flow cytometry 

分 类 号：O657.3[理学—分析化学] R698.2[理学—化学]