The influence of cell morphology on microfluidic single cell analysis

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 in situ 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.