The throughput of cell mechanical characterization has recently approached that of conventional flow cytometers. However, this very sensitive, label-free approach still lacks the specificity of molecular markers. Here we developed an approach that combines real-time 1D-imaging fluorescence and deformability cytometry in one instrument (RT-FDC), thus opening many new research avenues. We demonstrated its utility by using subcellular fluorescence localization to identify mitotic cells and test for mechanical changes in those cells in an RNA interference screen.

Highlights d Ras-ERK signaling controls cell shape in mitosis d Activation of Ras alters cell mechanics to enhance mitotic stiffening d Cells expressing Ras are better able to round up to divide under stiff gels d Ras limits DNA segregation errors normally observed in confined cell division