摘要
目的建立实用的实验程序，观察不同功能状态下的精子表面超微结构特点。方法用ＢＷＷ上游法分离活动的人精子：ＢＷＷ体外获能、诱导顶体反应和低渗膨胀试验处理后，将精子贴附于阳离子膜玻片上，叔丁醇冷却干燥后真空铂－碳喷镀复型。复型膜经３％氢氟酸剥脱，次氯酸钠腐蚀，透射电镜观察。结果人精子表面复型图像清晰，立体感强，细节较丰富、清晰，获能精子顶体区呈网络状，顶体反应时可见顶体区大量互相吻合的小泡，经低渗处理的精子顶体区出现类似获能和顶体反应的现象。结论表面复型－透射电镜技术可用于观察精子的不同生理状态引起的表面形貌变化，方法简便，可重复性好，具有良好的应用价值。
Objective\ To assess the variability of the surface replica method for the observation of functional ultrastructure of human sperm.Methods\ Motile human spermatozoa,separated by swimming up in BWW,before/after capacitation,induced acrosome reaction and pretreated with hypoosmotic medium (HOS test),were fixed,layered on the poly L lysine coated slide cover glass and dehydrated by tert butanol method.The sperms on cover glass were coated by Pt/C shadowing in high vacuum to form surface replica film.The film was stripped from glass by 3% hydrofluoric acid and digested with NaClO and then,after drying,was observed under transmission electron microscope.Results\ The images of surface replica of human sperm were clear and stereo to show the surface geomorphology of spermatozoa in different physiological state.Conclusion\ The method of surface replica can be used to analyze the functional altervation of sperm in ultrastructural level.
出处
《解剖学报》
CAS
CSCD
北大核心
1999年第2期181-183,I018,共4页
Acta Anatomica Sinica
关键词
表面复型
电子显微镜
获能
顶体反应
精子
Surface replica
Electron microscopy
Capacitation
Acrosome reaction
Spermatazoon
