Cytomics FC500与ImageStream^X MarkⅡ检测大鼠脾脏T、B细胞亚群TDO_2表达的比较

Comparison of TDO2 expression in T and B cell subsets of spleen detected by Cytomics FC500 and ImageStream^X MarkⅡ in rats

用常规方法制备大鼠脾脏细胞悬液,分别用Cytomics FC500流式细胞仪(FC500)和ImageStream^X MarkⅡ成像流式细胞仪(MarkⅡ)检测大鼠脾脏辅助性T细胞(CD3^+CD4^+)、细胞毒性T细胞(CD3^+CD8^+)、B细胞(CD45R^+)色氨酸2,3双加氧酶(TDO2)的表达水平,并观察2种检测技术检测结果的相关性和一致性。结果显示,2种流式细胞仪测得的CD3^+CD4^+T细胞、CD3^+CD8^+T细胞和CD45R^+B细胞表达TDO2的相关性较好(R^2均>0.8)。Bland-Altman法分析2种检测技术测得CD3^+CD4^+细胞、CD3^+CD8^+细胞和CD45R^+细胞表达TDO2的偏倚较小,分别为-1.3%、-8.2%、7.4%。TDO2在大鼠脾脏T、B细胞中均有表达。FC500和MarkⅡ检测结果的相关性和一致性较好。FC500操作方便,速度快。MarkⅡ将传统流式细胞术的高通量分析与细胞形态学分析相结合,精确度高,可获得细胞具体图片。

Rat spleen cell suspension was prepared by common method.The expression levels of helper T cells(CD3+CD4+),cytotoxic T cells(CD3+CD8+),and B cells(CD45R+)subsets tryptophan 2,3-dioxygenase(TDO2)in rat spleen were detected by Cytomics FC500(FC500)and ImageStreamX MarkⅡ(MarkⅡ)flow cy-tometry.The correlation and consistency of the results were observed respectively.The expression of TDO2 in CD3+CD4+T cells,CD3+CD8+T cells and CD45R+B cells were measured by two kinds of flow cytometry,and the cor-relation were well(R2>0.8).Bland-Altman method was used to analyze TDO2 expression in CD3+CD4+T cells,CD3+CD8+T cells and CD45R+B cells.The bias were-1.3%,-8.2%and 7.4%,respectively.TDO2 were expressed in T and B cells of rat spleen.These two flow cytometries had good correlation and consistency in testing resualts.FC500 is easy and fast to operate.Mark-Ⅱcombines the high-throughput analysis of traditional flow cy-tometry with cell mor-phology analysis,which is highly accurate and can obtain specific cell pictures.