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A comparison of methods to assess cell mechanical properties


The mechanical properties of cells influence their cellular and subcellular functions, including cell adhesion, migration, polarization, and differentiation, as well as organelle organization and trafficking inside the cytoplasm. Yet reported values of cell stiffness and viscosity vary substantially, which suggests differences in how the results of different methods are obtained or analyzed by different groups. To address this issue and illustrate the complementarity of certain approaches, here we present, analyze, and critically compare measurements obtained by means of some of the most widely used methods for cell mechanics: atomic force microscopy, magnetic twisting cytometry, particle-tracking microrheology, parallel-plate rheometry, cell monolayer rheology, and optical stretching. These measurements highlight how elastic and viscous moduli of MCF-7 breast cancer cells can vary 1,000-fold and 100-fold, respectively. We discuss the sources of these variations, including the level of applied mechanical stress, the rate of deformation, the geometry of the probe, the location probed in the cell, and the extracellular microenvironment.

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This research was supported by the NIH (grants U54CA143868 and R01CA174388 to D.W. and P.-H.W.; GM072744 to N.W.; GM096971 and CA193417 to P.A.J.; and CA143862 to R.R.), the NSF (grant 1510700 to R.R.), Agence Nationale de la Recherche (“ImmunoMeca” ANR-12-BSV5-0007-01, “Initiatives d’excellence” Idex ANR-11-IDEX-0005-02, and “Labex Who Am I?” ANR-11-LABX-0071 to A.A.), and the Deutsche Forschungsgemeinschaft through the collaborative research center (SFB1027 to A.O.).

Author information

A.A., J.G., P.A.J., A.O., R.R., I.S., N.W., D.W., J.S.H.L., and N.M.M. designed the study. P.-H.W., D.R.-B.A., W.-C.C., M.E.D., B.L.D., P.D.-S., A.E., N.V.G., Y.-C.P., M.S., J.R.S., and G.W. performed the experiments and analysis. P.-H.W., A.A., J.G., P.A.J., A.O., R.R., I.S., N.W., and D.W. wrote the manuscript.

Competing interests

The authors declare no competing interests.

Correspondence to Pei-Hsun Wu or Atef Asnacios or Jochen Guck or Paul A. Janmey or Albrecht Ott or Robert Ros or Igor Sokolov or Ning Wang or Denis Wirtz.

Integrated supplementary information

Supplementary Figure 1 Relaxation and creep functions of individual cells with the parallel-plate rheometer.

a, Typical creep function obtain for MCF 7 cell. b, Typical relaxation function obtain for MCF 7 cell. c, diagram representing the mean of the extensional modulus at 1Hz obtain for the different tests performed on MCF 7 cells with the parallel plates technique. (n=18 for the oscillation test, n=15 for the relaxation test, n =11 for the creep test). d, mean of the exponent of the power law found for the corresponding rheological tests in c. Error bars are standard errors.

Supplementary Figure 2 Further analysis and details of OS results.

a Distribution of initial compliance Jo for each MCF7 cell stretched (n = 514), based on the power law model. The dotted line represents the cumulative distribution. b Distribution of the power law exponent β. The average β here was found to be 0.85 ± 0.03. c. Average compliance curve for 11 MCF7 cells stretched using 1.5 W per fibre, showing more typical viscoelastic features than the cells stretched at 0.7 W per fibre as in the main text. d Distribution of the average refractive index obtained for 89 cells. Here, the population average is 1.374 ± 0.002.

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Supplementary Figures 1 and 2, Supplementary Notes 1–3 and Supplementary Tables 1 and 2

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Fig. 1: Description of rheological tests.
Fig. 2: AFM measurements.
Fig. 3: Whole-cell deformation measurements.
Fig. 4: Cell monolayer rheology.
Fig. 5: Bead-based measurements.
Supplementary Figure 1: Relaxation and creep functions of individual cells with the parallel-plate rheometer.
Supplementary Figure 2: Further analysis and details of OS results.