为比较外周血T淋巴细胞亚群CD4不同测定方法的差别,以流式细胞术为定量手段,测定了猴外周血中三种不同方法处理后CD4的表达.结果表明:先标后溶法——先用异硫氰荧光素标记的单克隆抗体(FITC-CD4 McAb)标记后,再加入红细胞溶解液溶掉红细胞的处理方法,结果基本等同于传统的淋巴细胞分离法,但样本用量仅为传统方法的1/5,且操作简单.激光共焦显微术的形态学研究也证实:先标后溶法与淋巴细胞分离法相似,其细胞膜表面荧光标记清晰,优于先溶后标法.
To investigate the differentiation of pre-preparation method to determine CD4 on T lymphocyte in blood, CD4 expression on T lymphocyte of monkey was determined by the flow cytometrey as a quantitative analysis with three different pre-preparation methods. These results showed that (b) method (whole blood was labeled by FITC-CD4 McAb. Then red blood cells were dissolved by erythrocytolysis) was better (a) method (red blood cells were dissolved by erythrocytolysis in whole blood. Then it was labeled by FITC-CD4 McAb). CD4 results with (b) method were equal to (c) method (traditional method of lymphocyte separated). But the volume of sample by (b) method only was 1/5 of (c) method. The pre-preparation of (b) method also was sampler than (c) method.Morphologic study by laser scanning confocal microscopy showed (b) method was similar to (c) method. Both of them had a clear fluorescent labeled on the cell membrane but not in the cell.
韩梅,吕平,何其华,袁兰,马士良,陶家平. T细胞亚群CD4测定方法的研究[J].生物化学与生物物理进展,2000,27(2):218-220
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