不同抗原修复法对视网膜石蜡切片相关功能蛋白表达检测的影响  

The effects of varied antigen retrieval methods on immunohistochemistry of functional proteins in retinal paraffin section

作　　者：李颖[1] 胡运韬[1] 马汀[1] 马志中[1] 

机构地区：[1]北京大学第三医院眼科中心,100191

出　　处：《眼科研究》2010年第1期15-18,共4页Chinese Ophthalmic Research

基　　金：国家重点基础研究发展计划(973)资助(2007CB512203)

摘　　要：目的比较4种抗原修复法对视网膜组织抗原修复的效果。方法分别应用胰蛋白酶消化、高压加热、水浴加热和微波法对视网膜组织切片进行抗原修复,比较400例胞内视黄醛结合蛋白(CRALBP)、视紫红质和视蛋白免疫组织化学染色效果。结果4种方法对3种蛋白的阳性表达无影响(P>0.05),但在组织完整性和背景染色方面差异有统计学意义(P<0.01)。胰蛋白酶消化法效果稳定,抗原表达强,背景清晰,组织完整。而高压加热、水浴加热和微波修复导致组织脱失皱褶、细胞核龟裂,且背景染色强。结论在视网膜组织病理学检测中,胰蛋白酶消化法是较理想的抗原修复方法。Background Antigen retrieval method is the key of improving the successful rate of immunohistochemical assay in paraffin sections. To study an available method of antigen retrieval is a goal to achieve both good immunoehemistry result and preserving retinal proteins. Objective The aim of present study is to investigate tyrosin digestion,high-temperature heat pressure, water bath heating and microwave repair in antigen retrieval for retinal tissues. Methods Retinal tissue was isolated and obtained from clean Chinchilla rabbits. Four hundreds retinal paraffin sections were prepared. Four kinds of antigen retrieval methods for retinal tissue including tyrosin digestion, high-temperature heat pressure, water bath heating and microwave repair were used respectively. The depigmentated retinal paraffin section without antigen retrieval was used as control. The positive rates of expression of CRALBP, Rhodopsin and opsin proteins were evaluated and compared among four kinds of antigen retrieval methods by immunochemistry. Results CRALBP,Rhodopsin and opsin protein was positively expressed in cytoplasm of retinal pigment epithelial cells and the outer segment of photoreeeptor respectively. No significant difference was found in the positive expression rates of these three proteins among four kinds of antigen retrieval methods （ P 〉 0. 05 ） , but the differences in tissue integrity and background staining were statistically significant （ P 〈 0. 01 ）. The structural damage of retina included loss and pucker of scalera,crack of nucleus and abnormal background stain in high-temperature heat pressure method, water bath heating method and microwave retrieval method. However, stable CRALBP, Rhodopsin and opsin protein expression and strain effectiveness, clear background without unspecific staining and integrated tissue were seen in tyrosin digestion method. Conclusion During the clinical pathology analysis of retinal tissue,the application of tyrosin digestion in antigen retrieval could obtain a better effectiveness.

关 键 词：抗原修复 免疫组织化学 胞内视黄醛结合蛋白 视紫红质 视蛋白 视网膜 

分 类 号：R774.1[医药卫生—眼科]