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

Abstract

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.

Single Cell Mechanotype and Associated Molecular Changes in Urothelial Cell Transformation and Progression

“…To verify this finding in a disease-specific setting, we integrated cell mechanotyping technology into clinical samples. Currently, a variety of platforms have been developed to measure cellular mechanical properties, and different techniques are known to probe mechanical properties at different timescales and depths of the cell (Urbanska et al, 2020). The most commonly explored method is to use biomechanical probes, represented by AFM and magnetic tweezer (Yang et al, 2011), whereas cell deformability can be optically probed under the force induced on the whole cells.…”

Engineering confining microenvironment for studying cancer metastasis

“…The mechanical properties of CTCs, in a way, reflect the resilience of cancer cells to the mechanical stresses, as well as their metastatic efficiency in circulation (Alibert et al, 2017;Wirtz et al, 2011). The mechanical characterization of cancer cells has led to an emerging field of mechanophenotyping that aims to elucidate the differences of mechanical properties between cancer and healthy cells for diagnostic and prognostic purposes (Urbanska et al, 2020;Wu et al, 2018). Traditionally, methods developed for the mechanical characterization of cells (Wu et al, 2018) include atomic force microscopy (Li et al, 2008), optical tweezers (Dao et al, 2003), micropipe aspiration (Mak and Erickson, 2013), etc.…”

Single-Cell Mechanophenotyping in Microfluidics to Evaluate Behavior of U87 Glioma Cells

“…Mechanical properties of cells, being closely linked to homeostasis of cells and their own microenvironment, have been hallmarks for defining healthy and malignant conditions of cells particularly in metastatic cancer [ 11 , 12 , 13 , 14 ]. Several different technologies have been developed to measure mechanical properties of cells including flow cytometry [ 15 , 16 , 17 ], atomic force microscopy (AFM) [ 18 , 19 ], magnetic twisting cytometry (MTC) [ 20 ], parallel-plate rheometry [ 11 , 21 ], optical stretching (OS) [ 22 ], optical tweezer [ 23 , 24 ], microfluidic ektacytometry [ 25 , 26 ], and micropipette aspiration [ 27 , 28 ]. Utilizing these modern tools for delineating mechanophenotypic properties of cells including stiffness, adhesiveness, viscosity, deformation (ratio of the area to volume), morphology, and migration trajectories of cells have been extensively investigated in cancer biology [ 29 , 30 , 31 ].…”