成像流式细胞术在检测巨噬细胞吞噬肿瘤细胞中的应用

Application of imaging flow cytometer in detecting tumor cell phagocytosis

分别用Cytomics FC500型流式细胞仪和ImageStreamX MarkⅡ成像流式细胞仪检测小鼠来源RAW264.7细胞转染对照组NC mimics和微小RNA-22(miR-22)mimics之后吞噬鼠源胶质瘤细胞系GL261的百分比,并观察两种检测技术检测结果的相关性和一致性。结果显示,两种流式细胞术测得的RAW264.7细胞(NC mimics组和miR-22 mimics组)吞噬GL261细胞的比例的相关性良好(R 2>0.8),两者结果差异有统计学意义(P<0.05)。Bland-Altman法分析两种检测技术测得RAW264.7细胞(NC mimics组和miR-22 mimics组)吞噬GL261细胞的比例偏倚较小,分别为4.289%和8.073%。与对照组NC mimics比较,miR-22 mimics组RAW264.7细胞吞噬肿瘤细胞能力增加(P<0.05),成像流式细胞术还可获得吞噬过程的单个细胞图片。经典流式细胞术操作方便,速度较快,而成像流式细胞术建立在传统流式细胞术基础之上,实现了对细胞图像进行多参数量化分析,能获得全新的细胞形态统计。

Cytomics FC500 flow cytometer and ImageStreamX MarkⅡimaging flow cytometer were used to detect the percentage of mouse-derived RAW264.7 cells transfected with control group NC mimics and microRNA-22(miR-22)mimics to phagocytose mouse-derived glioma cells GL261,and the correlation and consistency of the two detection technologies were observed.The results showed that the ratio of RAW264.7 cells(NC mimics and miR-22 mimics)phagocytosing GL261 cells measured by two kinds of flow cytometry had a good correlation(R 2>0.8).The Bland-Altman method analyzed the two detection techniques and found that the proportion of RAW264.7 cells(NC mimics and miR-22 mimics)phagocytosing GL261 cells was less biased,which were 4.289%and 8.073%,respectively.Compared with the control group NC mimics,the ability of RAW264.7 cells in miR-22 mimics group to swallow tumor cells increased(P<0.05),and imaging flow cytometry could also obtain individual cell pictures of the phagocytic process.Classical flow cytometry is easy to operate and fast while imaging flow cytometry is based on traditional flow cytometry,which realizes multi-parameter quantitative analysis of cell images and can obtain new statistical data of cell morphology.