A comparison of microfluidic methods for high-throughput cell deformability measurements

The mechanical phenotype of a cell is an inherent biophysical marker of its state and function, with potential value in clinical diagnostics. Several microfluidic-based methods developed in recent years have enabled single-cell mechanophenotyping at throughputs comparable to flow cytometery. Here we present a highly standardized cross-laboratory study comparing three leading microfluidic-based approaches to measure cell mechanical phenotype: constriction-based deformability cytometry (cDC), shear flow deformability cytometry (sDC), and extensional flow deformability cytometry (xDC). We show that all three methods detect cell deformability changes induced by exposure to altered osmolarity. However, a dose-dependent deformability increase upon latrunculin B-induced actin disassembly was detected only with cDC and sDC, which suggests that when exposing cells to the higher strain rate imposed by xDC, other cell components dominate the response. The direct comparison presented here serves to unify deformability cytometry methods and provides context for the interpretation of deformability measurements performed using different platforms.