人精子2种扫描电镜样品制备方法的比较

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

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。

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 was 14.1 ±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.