Cell stiffness measurements have led to insights into various physiological and pathological processes1,2. Although many cellular behaviours are influenced by intracellular mechanical forces3,4,5,6 that also alter the material properties of the cell, the precise mechanistic relationship between intracellular forces and cell stiffness remains unclear. Here we develop a cell mechanical imaging platform with high spatial resolution that reveals the existence of nanoscale stiffness patterns governed by intracellular forces. On the basis of these findings, we develop and validate a cellular mechanical model that quantitatively relates cell stiffness to intracellular forces. This allows us to determine the magnitude of tension within actin bundles, cell cortex and plasma membrane from the cell stiffness patterns across individual cells. These results expand our knowledge on the mechanical interaction between cells and their environments, and offer an alternative approach to determine physiologically relevant intracellular forces from high-resolution cell stiffness images.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $16.58 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
All the data supporting the findings of this study are available within the article or its Supplementary Information files, or from the corresponding authors on reasonable request.
Plodinec, M. et al. The nanomechanical signature of breast cancer. Nat. Nanotechnol. 7, 757–765 (2012).
Stewart, M. P. et al. Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding. Nature 469, 226–230 (2011).
Mammoto, T., Mammoto, A. & Ingber, D. E. Mechanobiology and developmental control. Annu. Rev. Cell Dev. Biol. 29, 27–61 (2013).
Ingber, D. E. Mechanobiology and diseases of mechanotransduction. Ann. Med. 35, 564–577 (2003).
Houk, A. R. et al. Membrane tension maintains cell polarity by confining signals to the leading edge during neutrophil migration. Cell 148, 175–188 (2012).
Wirtz, D., Konstantopoulos, K. & Searson, P. C. The physics of cancer: the role of physical interactions and mechanical forces in metastasis. Nat. Rev. Cancer 11, 512–522 (2011).
Fletcher, D. A. & Mullins, R. D. Cell mechanics and the cytoskeleton. Nature 463, 485–492 (2010).
Zhu, C., Bao, G. & Wang, N. Cell mechanics: mechanical response, cell adhesion, and molecular deformation. Annu. Rev. Biomed. Eng. 2, 189–226 (2000).
Ingber, D. E. Cellular tensegrity revisited I. Cell structure and hierarchical systems biology. J. Cell Sci. 116, 1157–1173 (2003).
Oberleithner, H. et al. Potassium softens vascular endothelium and increases nitric oxide release. Proc. Natl Acad. Sci. USA 106, 2829–2834 (2009).
Rotsch, C. & Radmacher, M. Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Biophys. J. 78, 520–535 (2000).
Eghiaian, F., Rigato, A. & Scheuring, S. Structural, mechanical, and dynamical variability of the actin cortex in living cells. Biophys. J. 108, 1330–1340 (2015).
Raman, A. et al. Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. Nat. Nanotechnol. 6, 809–814 (2011).
Rigato, A., Rico, F., Eghiaian, F., Piel, M. & Scheuring, S. Atomic force microscopy mechanical mapping of micropatterned cells shows adhesion geometry-dependent mechanical response on local and global scales. ACS Nano 9, 5846–5856 (2015).
Wang, A., Vijayraghavan, K., Solgaard, O. & Butte, M. J. Fast stiffness mapping of cells using high-bandwidth atomic force microscopy. ACS Nano 10, 257–264 (2016).
Dong, M. D., Husale, S. & Sahin, O. Determination of protein structural flexibility by microsecond force spectroscopy. Nat. Nanotechnol. 4, 514–517 (2009).
Brangwynne, C. P. et al. Microtubules can bear enhanced compressive loads in living cells because of lateral reinforcement. J. Cell Biol. 173, 733–741 (2006).
Kumar, S. et al. Viscoelastic retraction of single living stress fibers and its impact on cell shape, cytoskeletal organization, and extracellular matrix mechanics. Biophys. J. 90, 3762–3773 (2006).
Wang, N., Butler, J. P. & Ingber, D. E. Mechanotransduction across the cell-surface and through the cytoskeleton. Science 260, 1124–1127 (1993).
Katoh, K., Kano, Y., Amano, M., Kaibuchi, K. & Fujiwara, K. Stress fiber organization regulated by MLCK and Rho-kinase in cultured human fibroblasts. Am. J. Physiol. Cell Physiol. 280, C1669–C1679 (2001).
Rossier, O. M. et al. Force generated by actomyosin contraction builds bridges between adhesive contacts. EMBO J. 29, 1055–1068 (2010).
Théry, M., Pepin, A., Dressaire, E., Chen, Y. & Bornens, M. Cell distribution of stress fibres in response to the geometry of the adhesive environment. Cell Motil. Cytoskeleton 63, 341–355 (2006).
Deguchi, S., Ohashi, T. & Sato, M. Evaluation of tension in actin bundle of endothelial cells based on preexisting strain and tensile properties measurements. Mol. Cell Biomech. 2, 125–133 (2005).
Ghibaudo, M. et al. Traction forces and rigidity sensing regulate cell functions. Soft Matter 4, 1836–1843 (2008).
Tinevez, J. Y. et al. Role of cortical tension in bleb growth. Proc. Natl Acad. Sci. USA 106, 18581–18586 (2009).
Mahaffy, R. E., Park, S., Gerde, E., Käs, J. & Shih, C. K. Quantitative analysis of the viscoelastic properties of thin regions of fibroblasts using atomic forcemicroscopy. Biophys. J. 86, 1777–1793 (2004).
Fabry, B. et al. Time scale and other invariants of integrative mechanical behavior in living cells. Phys. Rev. E 68, 041914 (2003).
Bar-Ziv, R., Tlusty, T., Moses, E., Safran, S. A. & Bershadsky, A. Pearling in cells: a clue to understanding cell shape. Proc. Natl Acad. Sci. USA 96, 10140–10145 (1999).
Diz-Munoz, A., Fletcher, D. A. & Weiner, O. D. Use the force: membrane tension as an organizer of cell shape and motility. Trends Cell Biol. 23, 47–53 (2013).
Gao, H., Qian, J. & Chen, B. Probing mechanical principles of focal contacts in cell–matrix adhesion with a coupled stochastic–elastic modelling framework. J. R. Soc. Interface 8, 1217–1232 (2011).
Cunningham, C. C. et al. Actin-binding protein requirement for cortical stability and efficient locomotion. Science 255, 325–327 (1992).
Su, J., Muranjan, M. & Sap, J. Receptor protein tyrosine phosphatase alpha activates Src-family kinases and controls integrin-mediated responses in fibroblasts. Curr. Biol. 9, 505–511 (1999).
Sahin, O., Magonov, S., Su, C., Quate, C. F. & Solgaard, O. An atomic force microscope tip designed to measure time-varying nanomechanical forces. Nat. Nanotechnol. 2, 507–514 (2007).
Sahin, O. & Erina, N. High-resolution and large dynamic range nanomechanical mapping in tapping-mode atomic force microscopy. Nanotechnology 19, 445717 (2008).
Smith, M. B. et al. Segmentation and tracking of cytoskeletal filaments using open active contours. Cytoskeleton 67, 693–705 (2010).
We acknowledge M. P. Sheetz for helpful discussions, E. Bryant for help in setting up imaging and cell culture, T. Iskratsch and V. Stevenin for help with fibroblast culture and treatment, X. Chen for help with instrumentation, T. P. Stossel and F. Nakamura for providing M2 cells, L. Chasin for providing CHO cells and L. P. Alonso-Sarduy for culturing them. This work is supported by the NIH Director’s New Innovator Award Program (1DP2-EB018657), the Rowland Fellows Program and the Wyss Institute for Biologically Inspired Engineering at Harvard University.
O.S. is a co-inventor on US Patents US7302833B2 and US7404314B2 assigned to Stanford University, which are related to the methods used this work. O.S. founded Big Apple Nano, Inc.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.