Zhang Xuxin, Li Yanzhao, Fang Hanshu, Wei Hongquan, Mu Ying, Lang Ming-Fei, Sun Jing
Affiliated Zhongshan Hospital of Dalian University Dalian 116001 China.
Medical College, Institute of Microanalysis, Dalian University Dalian 116622 China
RSC Adv. 2018 Dec 21;9(1):139-144. doi: 10.1039/c8ra08303g. eCollection 2018 Dec 19.
Microfluidics has been widely used in single cell analysis. Current protocols allow either spread or round cells to be analyzed. However, the contribution of cell morphology to single cell analysis has not been noted. In this study, four proteins (EGFR, PTEN, pAKT, and pS6) in the EGFR signaling pathway are measured simultaneously using microfluidic image cytometry (MIC) in glioblastoma cells U87. The results show that the MIC technology can reveal different subsets of cells corresponding to the four protein expression levels no matter whether they are round or spread at the time of the measurements. However, sharper distinction is obtained from round cells, which implies that cellular heterogeneity can be better resolved with round cells during protein quantification by imaging cytometry. This study calls attention to the role of cell morphology in single cell analysis. Future studies should examine whether differences in data interpretation resulting from cell morphology could reveal altered biological meanings.
微流控技术已广泛应用于单细胞分析。目前的实验方案允许对铺展型或圆形细胞进行分析。然而，细胞形态对单细胞分析的影响尚未得到关注。在本研究中，利用微流控图像细胞术（MIC）在胶质母细胞瘤U87细胞中同时检测了表皮生长因子受体（EGFR）信号通路中的四种蛋白质（EGFR、PTEN、磷酸化AKT和磷酸化S6）。结果表明，无论在测量时细胞是圆形还是铺展型，MIC技术都能揭示与这四种蛋白质表达水平相对应的不同细胞亚群。然而，从圆形细胞中能获得更清晰的区分，这意味着在成像细胞术进行蛋白质定量分析时，圆形细胞能更好地解析细胞异质性。本研究提醒人们关注细胞形态在单细胞分析中的作用。未来的研究应探讨细胞形态导致的数据解读差异是否能揭示生物学意义的改变。
