精子处理方法对免疫荧光检测抗精子单克隆抗体的影响

THE INFLUENCES OF DIFFERENT TREATMENT METHODS OF SPERMATOZOA ON IMMUNOFLUORESCENCE DETERMINATION OF ANTISPERMATOZOON MONOCLONAL ANTIBODY

本文比较了用活精子、精子涂片不加固定及不同固定方法对间接免疫荧光检测效果的影响和精子抗原性保存问题。应用免疫荧光法筛选分泌抗精子单克隆抗体杂交瘤细胞系,各种精子处理方法均可采用。但用活精子常因操作中精子丢失而失败;检测与精子尾部结合的单克隆抗体,最好不加固定;检测与精子头部结合的单克隆抗体,用丙酮或4℃乙醇固定为佳;涂片不加固定抗原性易改变,而固定后其抗原性可保持6个月以上。

The influences of different treatment methods such as living spermatozoon, spermatozoa smear without fixation and different fixing methods on determination effect of immunofluorescence (indirect method) and the conservation of spermatozoon antigenicity are compared. Immunofiuorescent method is applied to select hybridoma cell system which secretes antispermatozoon monoclonal antibody, all kinds of methods of spermatozoon treatment can be used.Although atrong fluorescence can be obtained by us- ing living spermatoza, but it is often to fail because of the spermia loss in procedure.To de- termine monoclonal antibody combined with the sperm tail, it is better that the smear is not fixed. However, to determine monoclonal antibody combined with the sperm head, the favourable way is to fix it with acefone or 4℃ ethyl alcohol.If the smear is not fixed, antigenicity of spermatozoa is easy to change, and the antigenicity can be kept for more six monthes after fialng.