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目的:通过建立自制量子点荧光免疫组织化学技术,对人晶状体缝隙连接蛋白(Connexin,Cx)50表达水平进行检测。方法:正常人和先天性白内障患者的晶状体制备石蜡切片,将羧基修饰的量子点和鼠抗兔的二抗相结合,分别采用链霉卵白素-过氧化物酶连结(SP)法和量子点偶联的二抗SP法对正常人和先天性白内障患者的晶状体细胞Cx50蛋白表达水平进行检测。结果:先天性白内障患者晶状体细胞Cx50蛋白表达水平高于正常对照组,量子点偶联的二抗SP法与常规SP法相比,具有敏感性高的优点。结论:量子点荧光标记二抗在检测组织蛋白表达水平中具有一定优势,值得推广。
OBJECTIVE: To detect the expression of connexin (Cx) 50 in human lens by establishing self-made quantum dots fluorescence immunohistochemistry. Methods: Paraffin sections were prepared from the lens of normal and congenital cataract patients. The carboxy - modified quantum dots were combined with the secondary antibody of the mouse anti - rabbit, and then were detected by streptavidin - peroxidase (SP) Coupled secondary antibody SP method in normal and congenital cataract patients lens cells Cx50 protein levels were detected. Results: The expression of Cx50 protein in lens cells of patients with congenital cataract was higher than that of the normal control group. Quantum dots-coupled secondary antibody SP method had the advantages of high sensitivity compared with the conventional SP method. Conclusion: Quantum dot fluorescent labeled secondary antibody has certain advantages in the detection of tissue protein expression level, which deserves promotion.