Objective To explore the effects of centrifugal forces on CD45 expression detection by mass cytometry in colon cancer cells.Methods The colon cancer HCT116 cells were divided into two groups by centrifugal forces as follows: 400 g group and 140 g group (3×106 cells in each group), and we used 400 g and 140 g centrifugal forces for 5 min, twice, to collect the cells.The percentage of dead cells, cell surface antigen CD45 expression, and live cell percentage of 154Sm signal intensity over than 102, were tested by mass cytometry in each group.Results In the 400 g group and 140 g group, the percentages of dead cells were 19.06%±0.66% and 7.69%±0.7%, respectively (P<0.01);the cell percentage of 154Sm signal intensity over than 102 was 2.9% (2.88%±0.03%) in the 400 g group, but only 0.2% (0.19%±0.08%) in the 140 g group (P<0.01).t-SNE results showed there were were 167.41±70.48 and 201.20±20.96 cells with high expression of CD45 in 400 g group and 140 g group, respectively (P<0.01).Conclusion Our results show that 140 g centrifugal force is better than 400 g in detection of CD45 expression in colon cancer cells by mass cytometry.%目的 探讨离心力对质谱流式细胞技术检测结肠癌细胞表面抗原CD45表达结果的影响.方法 将体外培养的结肠癌HCT116细胞分为400 g离心组和140 g离心组(每组细胞数均为3×106个),分别采用400 g、140 g离心力离心5 min,重复两次,收集细胞.采用质谱流式细胞技术检测两组死亡细胞百分比、抗人CD45金属聚合抗体(154Sm)信号强度>102的细胞数占总活细胞数的比例、t-SNE图显示活细胞中高表达CD45的细胞数量.结果 400 g离心力组及140 g离心力组死亡细胞百分比分别为(7.69±0.7)%、(19.06±0.66)%,两组比较P<0.01.400 g离心力组及140 g离心力组154Sm信号强度>102的细胞数分别占活细胞总数的0.2%[(0.19±0.08)%]、2.9%[(2.88±0.03)%],两组比较P<0.01.t-SNE图显示,400 g离心力组及140 g离心力组CD45高表达细胞分别为(167.41±70.48)、(201.20±20.96)个,两组比较P<0.01.结论 应用质谱流式细胞技术检测结肠癌细胞表面抗原CD45表达时,140 g离心力分离的细胞检测结果更好.
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