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A comparison of methods to assess cell mechanical properties

Nature Methods volume 15, pages 491–498 (2018)Cite this article

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Abstract

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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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.

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Acknowledgements

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.).

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering and Departments of Pathology and Oncology, The Johns Hopkins University and Johns Hopkins School of Medicine, Baltimore, MD, USA

    Pei-Hsun Wu, Wei-Chiang Chen, Jerry S. H. Lee & Denis Wirtz

  2. Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, PA, USA

    Dikla Raz-Ben Aroush & Paul A. Janmey

  3. Laboratoire Matière et Systèmes Complexes, Unité Mixte de Recherche 7057, Centre National de la Recherche Scientifique (CNRS) and Université Paris–Diderot (Paris 7), Sorbonne Paris Cité, Paris, France

    Atef Asnacios & Pauline Durand-Smet

  4. Department of Mechanical Engineering, Tufts University, Medford, MA, USA

    Maxim E. Dokukin & Igor Sokolov

  5. Department of Physics, Arizona State University, Tempe, AZ, USA

    Bryant L. Doss, Robert Ros & Jack R. Staunton

  6. Biotechnology Center, Technische Universität Dresden, Dresden, Germany

    Andrew Ekpenyong, Jochen Guck & Graeme Whyte

  7. Department of Physics, Clarkson University, Potsdam, NY, USA

    Nataliia V. Guz

  8. Center for Strategic Scientific Initiatives, National Cancer Institute, Bethesda, MD, USA

    Jerry S. H. Lee

  9. Department of Mechanical Science and Engineering, College of Engineering, University of Illinois at Urbana–Champaign, Urbana, IL, USA

    Nicole M. Moore, Yeh-Chuin Poh & Ning Wang

  10. Biological Experimental Physics Department, Saarland University, Saarbruecken, Germany

    Albrecht Ott & Mathias Sander

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Contributions

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.

Corresponding authors

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

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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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Wu, PH., Aroush, D.RB., Asnacios, A. et al. A comparison of methods to assess cell mechanical properties. Nat Methods 15, 491–498 (2018). https://doi.org/10.1038/s41592-018-0015-1

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