人精子2种扫描电镜样品制备方法的比较

目的:比较人精子2种扫描电镜样品制备方法的效果。方法:10例精液标本,同一例标本分别采用扫描电镜一般样品制备程序(方法A,A组)和本实验室方法(方法B,B组)处理。方法:B组减少了固定后洗涤精子、脱水和醋酸异戊脂置换的时间,增加了多级乙醇浓度梯度。观察2组开裂精子率的差别。结果:应用2种标本制备程序处理的各例标本,均可观察到开裂或断裂的精子。裂开部位可发生在精子头部、颈部或尾部,以发生在头部为多见,其次为颈部。A组的开裂精子率为28.9±16.2%,B组的为14.1±7.7%,B组的显著少于A组(P<0.01)。结论:方法B的程序更适合于人精子标本的制备,可减少精子表面的开裂或结构断裂,有利于人精子超微形态学观察。

Comparison of Two Preparations of the Scanning Electron Microscopy for Human Sperm

Objective:To compare the quality of two preparations of the scanning electron microscopy(SEM)for human sperm.Methods:Ten semen samples were used to evaluate the two preparations(method A,and method B)of the SEM.Method A was a routine preparation of the SEM for cells.Method B was the modified preparation of our laboratory,in which time for washing sperm after fixative,dehydration,and replacing agent were reduced,graded ethanols were increased.The incidences of ruptured sperm between methods A and B were compared.Results:Following the two preparations,ruptured sperm were found in all 10 samples under SEM.The ruptured head of sperm was common.The rate of ruptured sperm in method B was14.1±7.7%,and was significant lower than that in method A(28.9 ± 16.2%)(P<0.01).Conclusion:The modified method B could reduce the ruptured sperm,which could improve the quality of the preparation of SEM for human sperm.