A Systematic Study of Size Correlation and Young's Modulus Sensitivity for Cellular Mechanical Phenotyping by Microfluidic Approaches.

Cellular mechanical properties are a class of intrinsic biophysical markers for cell state and health. Microfluidic mechanical phenotyping methods have emerged as promising tools to overcome the challenges of low throughput and high demand for manual skills in conventional approaches. In this work, two types of microfluidic cellular mechanical phenotyping methods, contactless hydro-stretching deformability cytometry (lh-DC) and contact constriction deformability cytometry (cc-DC) are comprehensively studied and compared. Polymerized hydrogel beads with defined sizes are used to characterize a strong negative correlation between size and deformability in cc-DC (r = -0.95), while lh-DC presents a weak positive correlation (r = 0.13). Young's modulus sensitivity in cc-DC is size-dependent while it is a constant in lh-DC. Moreover, the deformability assessment for human breast cell line mixture suggests the lh-DC exhibits better differentiation capability of cells with different size distributions, while cc-DC provides higher sensitivity to identify cellular mechanical changes within a single cell line. This work is the first to present a quantitative study and comparison of size correlation and Young's modulus sensitivity of contactless and contact microfluidic mechanical phenotyping methods, which provides guidance to choose the most suitable cellular mechanical phenotyping platform for specific cell analysis applications.