考马斯亮蓝染色法检测人精子顶体反应的评价

为建立一种检测人精子顶体反应的简便实用的新技术，用３．５％高氯酸水溶液配制０．０５％考马斯亮蓝（Ｒ２５０）染色液浸染人精子３０ｍｉｎ，顶体完整者顶体区染成紫蓝色，顶体反应者则不染。对获能前后和钙离子载体Ａ２３１８７诱导顶体反应的精子进行染色并与经典的顶体反应检测技术ＰＳＡ法对照，两种方法检测顶体反应率无显著差异。方法学检验证明新技术结果可靠。本法操作简便、经济，普通显微镜即可检测，易于推广，克服了原有技术复杂昂贵的缺点，具有研究和临床诊断的良好价值。

EVALUATION OF COOMASSIE BRILLIANT BLUE STAINING FOR DETECTION OF HUMAN SPERM ACROSOME REACTION

The detection of human sperm acrosome reaction (AR) Coomassie brilliant blue (CBB)staining is evaluated in the present research. The technique is a modification from Aaron's method on mouse(Aarons D et al,1991).The smear of prefixed spermatozoa after swimming up, capacitation or A23187 pretreatment were stained for 30 min with 0.05% coomassie brilliant blue R250 which resolved in 3 5% perchloric acid (HClO 4) and visualized with light microscopy. The sperm population was divided by CBB staining into two types:type A and type B. Type A was positive stained with dark violet blue on acrosome cap while type B was pale or negative stained on the same area. The percentage of type B sperm increased significantly after capacitation or AR induction. A comparison study showed there is no difference in assessing AR rate between CBB method and conventional PSA technique. Therefore, the simple and economical CBB staining procedure provides a reliable method for assessment of the human sperm acrosome reaction.