应用免疫组化法验证细胞芯片的特异性  

Study of the Immunohistochemistry on the Cell Microarray

作　　者：胥威[1] 赵雨杰[1] 卢永平[1] 宋瑾[2] 陈亚男[2] 李春丽[2] 徐晓雪[1] 

机构地区：[1]中国医科大学基础医学院生物芯片中心,沈阳110001 [2]中国医科大学基础医学院细胞生物学教研室,沈阳110001

出　　处：《中国医科大学学报》2008年第4期491-493,共3页Journal of China Medical University

基　　金：辽宁省教育厅高等学校科研基金资助项目(202011001)

摘　　要：目的研究适用于芯片捕获细胞的最佳免疫组化方法,并用其验证细胞芯片的特异性。方法CD7、CD13、CD19抗体固定于醛基玻片上,形成3×3阵列的芯片,从外周血中分离全白细胞,芯片上孵育后,细胞芯片捕获的细胞固定,S-P法进行免疫组化及苏木素复染,显微镜下观察和计数。结果乙醇∶甲醛∶水(AF液)固定胞内外抗原染色阳性率、阳性程度高,阳性细胞形态较好;芯片上免疫组化所用抗体最佳浓度为1μg/ml,最佳孵育条件为室温,1h;芯片上阳性细胞胞浆或胞膜有棕色颗粒,阴性抗体点细胞无此表现;细胞芯片的特异性结合阳性率在96.7%以上,非特异性结合阳性率在0.8%以下。结论本实验方法能将细胞芯片捕获细胞的胞内及胞浆抗原直观稳定表达出来,设计出了细胞芯片上免疫组化方法,同时验证了细胞芯片的特异性。Objective To research the best immunohistochemistry on the cell micmarray. Methods Monoclonal antibodies （MoAb） CD7, CD13 ,and CD19 were spotted on the aldehyde slide ,forming a 3×3 micmarray. The entire leucocytes, isolated from peripheral blood,were dripped on the slide. Different cellular groups can be combined with different antibodies. Leucocytes were fixed on the cell microarray, Their antigens presented with the streptavidin-pemxidase immunohistochemistry. The cell microarray were dyed with hematoxylin,observed and counted under the microscope. Results The positive rate and the positive degree of antigens on cell membrane and cytoplasm, fixed with the AF fluid,were the highest,the positive cellular forms are better; the best concentration of MoAb used in the immunohistochemistry on the cell micmarray is 1 μg/ml, the condition is room temperature for 1 hour; there are brown particles on cell membrane and in cytoplasm of the positive cells on the cell micmarray, but not on the negative cells. Conclusion This experiment could make antigens on cell membrane and in cytoplasm of the cells captured by the cell micmarray direct-viewingly and stably express, design immunohistochemica microarray, simultaneously confirm the reliability of cell micmarray.

关 键 词：细胞芯片 免疫组化 CD抗体 

分 类 号：R392.12[医药卫生—免疫学]