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目的 深入研究加热抗原修复对内源性抗生物素蛋白结合物 (EABA)的影响程度和范围 ,以及探讨消除EABA对免疫组织化学染色干扰的对策或方法。方法 采用高效、快捷的组织芯片技术、微波加热处理法、免疫组织化学SP染色法及蛋清液封闭法 ,对甲醛固定石蜡包埋的 76种 ( 10 2份 )人体正常组织和 88种 ( 5 77份 )人体肿瘤组织以及 4种 ( 80份 )大鼠正常组织标本进行系统性EABA检查 ,同时对 9种 ( 15份 )人体冷冻组织进行EABA检查。结果 ( 1)冷冻组织中存在EABA ;( 2 )经甲醛固定石蜡包埋后组织中的EABA被封闭 ;( 3 )加热抗原修复可造成EABA暴露 ;( 4)EABA暴露的强度各不相同 ,强者可到强阳性 ;( 5 )EABA在组织中的分布形式 ,既有散在分布也有弥漫分布 ,主要以颗粒状形式存在于胞质中 ;( 6)EABA广泛存在于上皮源性组织 ,特别是腺上皮组织 (包括正常组织和肿瘤组织 ) ,亦存在于部分非上皮组织 ;( 7)EABA暴露的强弱与修复液亦有关 ,EGTA(pH 9.0 )的暴露能力较柠檬酸 (pH 6.0 )和乙二胺四乙酸二钠 (pH 8.0 )更强 ;( 8)加热抗原修复暴露的EABA可以被 2 0 %蛋清液封闭。结论 加热修复可使甲醛固定石蜡包埋组织中的EABA重新暴露 ,暴露的EABA可广泛见于人体正常和肿瘤组织细胞中 ,也见于大鼠正常肝、肾、肺、胰组?
Objective To study in depth the extent and scope of the effect of heated antigen retrieval on endogenous avidin conjugates (EABA) and to explore ways and means to eliminate the interference of EABA on immunohistochemical staining. Methods Sixty-two (102) human paraffin-embedded normal tissues and 88 (577) human paraffin-embedded specimens were obtained by using efficient and rapid tissue microarray technique, microwave heating method, immunohistochemical staining and egg white solution. ) Human tumor and normal tissues of four kinds of rats (80) for EABA examination. EABA tests were performed on nine kinds of human frozen tissues (15). Results (1) EABA was present in the frozen tissue; (2) EABA was blocked in the paraffin-embedded paraffin-embedded tissue; (3) EABA exposure was induced by heating the antigen; (4) EABA exposure varied in intensity and strength (5) The distribution of EABA in the tissue, both scattered and diffuse distribution, mainly in granular form exists in the cytoplasm; (6) EABA is widely found in epithelial tissues, especially (7) The intensity of EABA exposure was also related to repair fluid, and the exposure capacity of EGTA (pH 9.0) was higher than that of citric acid (pH 6.0) and Disodium EDTA (pH 8.0) is stronger; (8) EABA exposed to heat antigen repair can be blocked with 20% egg white solution. Conclusion Heat repair can re-expose EABA in formalin-fixed paraffin-embedded tissues. Exposed EABA can be widely found in human normal and tumor cells, also in normal liver, kidney, lung and pancreatic tissues of rats.