改良常规SP免疫组化染色法的探讨

Dicussion of Enhancement of Normal SP Immunohistochemical Staining

目的探讨在固定液和抗体的稀释方法上做一些改进后对免疫组化染色结果的影响。方法:取成年雌性SD大鼠异位的子宫内膜,分别固定于4℃的丙酮、常温丙酮和10%甲醛固定液中,做常规石蜡包埋,切片,将3种固定液的切片分为两组,"一抗"分别采用倍比稀释、直接稀释的方法,做SP免疫组化染色实验比对。结果:4℃的丙酮作固定液与常温丙酮和10%甲醛处理的石蜡切片(经抗原修复)相比,抗原定位明确、阳性细胞表达率高,阳性细胞分布更均匀;抗体的倍比稀释常规直接稀释与对照,背景干净清晰。结论:用4℃的丙酮作固定液,"一抗"用倍比稀释的方法进行免疫组化染色,使得抗原定位明确、阳性反应与阴性背景对比清楚。

Objective To discuss the effect of the enhancement in method of stationary fluid and dilution of antibody. Methods Take ectopic endometrium from mature female SD rats, stabilized in 4℃ acetone, room temperature acetone, and 10% formalin as stationary fluid separately in each group. Use conventional paraffin - embedded, slicing to separate three groups above into two subgroups. Use direct dilute and double dilute in primary antibody, compare in SP immunohistochemical staining experiment. Result Room temperature acetone and 10% formalin , 4℃ acetone has cleaner and brighter background, clearer positive expression, and distributed more evenly in the paraffin section（ via antigen retrieval） ; Double dilution has more accurate pinpoint of antigen. It also has more clear positive reaction and negative background comparing to direct dilution. Conclusion： Using 4℃ acetone as stationary fluid and using double dilute in primary antibody for SP immunohistochemical staining makes better pinpointing antigen, and has more clear background and positive expression .