【作者】 秦海明； 赵雨杰； 侯伟建； 刘健；

【Author】 QIN Haiming, ZHAO Yujie, HOU Weijian, LIU Jian, Department of Clinical Laboratory, North Hospital, Shenyang, China, 110016,Biochip Center, China Medical University;Department of Biology, Shenyang Medical Collage.

【机构】 沈阳军区总医院检验科； 中国医科大学生物芯片中心； 沈阳医学院生物教研室；

【摘要】 目的本试验设计了一种全新概念的细胞芯片。利用抗原抗体特异性反应原理,将细胞膜表面标志CD抗原与玻片上固定的CD抗体进行反应,既保持了细胞的完整性及活性,又可以将不同细胞进行分群。方法将CD3,CD4,CD8抗体固定于醛基玻片上,形成微阵列,从全血中分离出的单个核细胞与之杂交,形成选择性细胞群。将细胞芯片进行胎盘兰染色,瑞氏染色,CD3-Cy5+CD4-FITC+CD8-RPE三色荧光染色并观察结果。结果该芯片能选择性的结合不同免疫表型的细胞。胎盘兰染色显示活细胞数>95%;瑞氏染色细胞形态良好,杂交点边缘整齐,背景干净;荧光染色显示特异性良好。结论本实验所设计的新型细胞芯片初步试验已达到预先要求。能根据细胞表面不同的免疫标志进行细胞分群,可以为以后制作大规模,高通量的细胞芯片做准备,确定了其可行性,并提出了全新意义的细胞芯片模型。更多还原

【Abstract】 Objective We developed a new notional cell microarray according antibody antigen reaction, which can keep the cellular integrality and alive and examined conveniently. Methods Antibodies of CD3, CD4 and CD8 were spotted on the aldehyde slide. Lymphocytes, isolated from peripheral blood (treated with helarin to prevent clotting) were hybridized with the slide. Different cellular groups can be combined with different antibodies. The cell microarrays were dyed with Trypan Blue, Wright’s dying and three colors fluorescence. Results A suspension of cells is applied to the array, and cells only bind to antibody dots for which they express the corresponding CD antigen. The binded cells were alive and had a good shape. The result of scan showed is right. Conclusion The technique of cell microarray is practicable because of its simple, rapid, low cost. Its results can be educed easily with a common optics microscope and also be scanned with a scanner to high throughput analysis.更多还原