Mechanobiology emerges at the crossroads of medicine, biology, biophysics and engineering and describes how the responses of proteins, cells, tissues and organs to mechanical cues contribute to development, differentiation, physiology and disease. The grand challenge in mechanobiology is to quantify how biological systems sense, transduce, respond and apply mechanical signals. Over the past three decades, atomic force microscopy (AFM) has emerged as a key platform enabling the simultaneous morphological and mechanical characterization of living biological systems. In this Review, we survey the basic principles, advantages and limitations of the most common AFM modalities used to map the dynamic mechanical properties of complex biological samples to their morphology. We discuss how mechanical properties can be directly linked to function, which has remained a poorly addressed issue. We outline the potential of combining AFM with complementary techniques, including optical microscopy and spectroscopy of mechanosensitive fluorescent constructs, super-resolution microscopy, the patch clamp technique and the use of microstructured and fluidic devices to characterize the 3D distribution of mechanical responses within biological systems and to track their morphology and functional state.
The versatile functions of biological systems ranging from molecules, cells and cellular systems to living organisms are governed by their mechanical properties and ability to sense mechanical cues and respond to them.
Atomic force microscopy (AFM)-based approaches provide multifunctional nanotools to measure a wide variety of mechanical properties of living systems and to apply to them well-defined mechanical cues.
AFM allows us to apply and measure forces from the piconewton to the micronewton range on spatially defined areas with sizes ranging from the sub-nanometre to several tens of micrometres.
Mechanical parameters characterized by AFM include force, pressure, tension, adhesion, friction, elasticity, viscosity and energy dissipation.
The mechanical parameters of complex biological systems can be structurally mapped, with a spatial resolution ranging from millimetres to sub-nanometres and at kinetic ranges from hours to milliseconds.
AFM can be combined with various complementary methods to characterize a multitude of mechanical, functional and morphological properties and responses of complex biological systems.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Trends in mechanobiology guided tissue engineering and tools to study cell-substrate interactions: a brief review
Biomaterials Research Open Access 01 June 2023
Mechanical stimulation devices for mechanobiology studies: a market, literature, and patents review
Bio-Design and Manufacturing Open Access 01 April 2023
High-resolution line-scan Brillouin microscopy for live imaging of mechanical properties during embryo development
Nature Methods Open Access 30 March 2023
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 digital issues and online access to articles
$99.00 per year
only $8.25 per issue
Rent or buy this article
Get just this article for as long as you need it
Prices may be subject to local taxes which are calculated during checkout
Hoffman, B. D., Grashoff, C. & Schwartz, M. A. Dynamic molecular processes mediate cellular mechanotransduction. Nature 475, 316–323 (2011).
Howard, J., Grill, S. W. & Bois, J. S. Turing’s next steps: the mechanochemical basis of morphogenesis. Nat. Rev. Mol. Cell Biol. 12, 392–398 (2011).
Brugues, A. et al. Forces driving epithelial wound healing. Nat. Phys. 10, 683–690 (2014).
Petridou, N. I., Spiro, Z. & Heisenberg, C. P. Multiscale force sensing in development. Nat. Cell Biol. 19, 581–588 (2017).
Tyler, W. J. The mechanobiology of brain function. Nat. Rev. Neurosci. 13, 867–878 (2012).
Crest, J., Diz-Munoz, A., Chen, D. Y., Fletcher, D. A. & Bilder, D. Organ sculpting by patterned extracellular matrix stiffness. eLife 6, e24958 (2017).
Heisenberg, C. P. & Bellaiche, Y. Forces in tissue morphogenesis and patterning. Cell 153, 948–962 (2013).
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).
Cattin, C. J. et al. Mechanical control of mitotic progression in single animal cells. Proc. Natl Acad. Sci. USA 112, 11258–11263 (2015). This paper shows that mechanical confinement by the atomic force microscope cantilever can be used to slow down and stop cells progressing through mitosis.
Roca-Cusachs, P., Conte, V. & Trepat, X. Quantifying forces in cell biology. Nat. Cell Biol. 19, 742–751 (2017).
Roos, W. H., Bruinsma, R. & Wuite, G. J. L. Physical virology. Nat. Phys. 6, 733–743 (2010).
Mateu, M. G. Structure and Physics of Viruses (Springer, Netherlands, 2013).
Fletcher, D. A. & Mullins, R. D. Cell mechanics and the cytoskeleton. Nature 463, 485–492 (2010).
Bieling, P. et al. Force feedback controls motor activity and mechanical properties of self-assembling branched actin networks. Cell 164, 115–127 (2016).
Bustamante, C., Chemla, Y. R., Forde, N. R. & Izhaky, D. Mechanical processes in biochemistry. Annu. Rev. Biochem. 73, 705–748 (2004).
Puchner, E. M. & Gaub, H. E. Force and function: probing proteins with AFM-based force spectroscopy. Curr. Opin. Struct. Biol. 19, 605–614 (2009).
Sauer, R. T. & Baker, T. A. AAA+ proteases: ATP-fueled machines of protein destruction. Annu. Rev. Biochem. 80, 587–612 (2011).
Iskratsch, T., Wolfenson, H. & Sheetz, M. P. Appreciating force and shape-the rise of mechanotransduction in cell biology. Nat. Rev. Mol. Cell Biol. 15, 825–833 (2014).
Alsteens, D. et al. Atomic force microscopy-based characterization and design of biointerfaces. Nat. Rev. Mater. 2, 17008 (2017).
Dufrene, Y. F. et al. Imaging modes of atomic force microscopy for application in molecular and cell biology. Nat. Nanotechnol. 12, 295–307 (2017).
Binnig, G., Quate, C. F. & Gerber, C. Atomic force microscope. Phys. Rev. Lett. 56, 930–933 (1986). This classical paper introduces the AFM technique.
Gerber, C. & Lang, H. P. How the doors to the nanoworld were opened. Nat. Nanotechnol. 1, 3–5 (2006).
Moeendarbary, E. et al. The cytoplasm of living cells behaves as a poroelastic material. Nat. Mater. 12, 253–261 (2013). The authors combine AFM with advanced microscopy to explain that the sponge-like behaviour of cells relates to a viscous fluid (cytosol) flowing through a porous elastic matrix (cytoskeleton, organelles and macromolecules).
Hecht, F. M. et al. Imaging viscoelastic properties of live cells by AFM: power-law rheology on the nanoscale. Soft Matter 11, 4584–4591 (2015).
Stewart, M. P. et al. Hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding. Nature 469, 226–230 (2011). This paper shows that to facilitate the drastic shape changes required for mitosis, animal cells generate hydrostatic pressure, which is balanced by a contracting actomyosin cortex.
Ramanathan, S. P. et al. Cdk1-dependent mitotic enrichment of cortical myosin II promotes cell rounding against confinement. Nat. Cell Biol. 17, 148–159 (2015).
Fabry, B. et al. Scaling the microrheology of living cells. Phys. Rev. Lett. 87, 148102 (2001). This paper represents a milestone in cellular rheology, as it introduces the notion of timescale-invariant mechanical properties and the power-law behaviour of cells.
Deng, L. et al. Fast and slow dynamics of the cytoskeleton. Nat. Mater. 5, 636–640 (2006).
Fischer-Friedrich, E. et al. Rheology of the active cell cortex in mitosis. Biophys. J. 111, 589–600 (2016).
Sen, S., Subramanian, S. & Discher, D. E. Indentation and adhesive probing of a cell membrane with AFM: theoretical model and experiments. Biophys. J. 89, 3203–3213 (2005).
Fischer-Friedrich, E., Hyman, A. A., Julicher, F., Muller, D. J. & Helenius, J. Quantification of surface tension and internal pressure generated by single mitotic cells. Sci. Rep. 4, 6213 (2014).
Vorselen, D., Kooreman, E. S., Wuite, G. J. & Roos, W. H. Controlled tip wear on high roughness surfaces yields gradual broadening and rounding of cantilever tips. Sci. Rep. 6, 36972 (2016).
Mahaffy, R. E., Shih, C. K., MacKintosh, F. C. & Kas, J. Scanning probe-based frequency-dependent microrheology of polymer gels and biological cells. Phys. Rev. Lett. 85, 880–883 (2000).
Toyoda, Y. et al. Genome-scale single-cell mechanical phenotyping reveals disease-related genes involved in mitotic rounding. Nat. Commun. 8, 1266 (2017). This paper presents a genome-scale AFM-based screen that discovers unexpected genes that animal cells use to round for mitosis.
Stewart, M. P. et al. Wedged AFM-cantilevers for parallel plate cell mechanics. Methods 60, 186–194 (2013).
Garcia, R. & Herruzo, E. T. The emergence of multifrequency force microscopy. Nat. Nanotechnol. 7, 217–226 (2012).
Dufrene, Y. F., Martinez-Martin, D., Medalsy, I., Alsteens, D. & Muller, D. J. Multiparametric imaging of biological systems by force-distance curve-based AFM. Nat. Methods 10, 847–854 (2013).
Eisenberg, B. R. & Mobley, B. A. Size changes in single muscle fibers during fixation and embedding. Tissue Cell 7, 383–387 (1975).
Dubochet, J. et al. Cryo-electron microscopy of vitrified specimens. Q. Rev. Biophys. 21, 129–228 (1988).
Hoh, J. H. & Schoenenberger, C. A. Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J. Cell Sci. 107, 1105–1114 (1994).
Meredith, J. E., Fazeli, B. & Schwartz, M. A. The extracellular-matrix as a cell-survival factor. Mol. Biol. Cell 4, 953–961 (1993).
Sader, J. E. & White, L. Theoretical analysis of the static deflection of plates for atomic force microscope applications. J. Appl. Phys. 74, 1–9 (1993).
Butt, H. J. & Jaschke, M. Calculation of thermal noise in atomic force microscopy. Nanotechnology 6, 1–7 (1995).
te Riet, J. et al. Interlaboratory round robin on cantilever calibration for AFM force spectroscopy. Ultramicroscopy 111, 1659–1669 (2011).
Schillers, H. et al. Standardized nanomechanical atomic force microscopy procedure (SNAP) for measuring soft and biological samples. Sci. Rep. 7, 5117 (2017). This paper introduces an approach to reducing the variability in extracting the elastic moduli of soft samples and reports on its validation in various laboratories to establish it as a standard method.
Wegmann, S., Medalsy, I. D., Mandelkow, E. & Muller, D. J. The fuzzy coat of pathological human Tau fibrils is a two-layered polyelectrolyte brush. Proc. Natl Acad. Sci. USA 110, E313–E321 (2013).
Vorselen, D., MacKintosh, F. C., Roos, W. H. & Wuite, G. J. Competition between bending and internal pressure governs the mechanics of fluid nanovesicles. ACS Nano 11, 2628–2636 (2017).
Hertz, H. Über die Berührung fester elastischer Körper [German]. Reine Angew. Math. 92, 156–171 (1881).
Sneddon, I. N. The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile. Int. J. Eng. Sci. 3, 47–57 (1965).
Johnson, K. L., Kendall, K. & Roberts, A. D. Surface energy and the contact of elastic solids. Proc. R. Soc. A 324, 301–313 (1971).
Rigato, A., Miyagi, A., Scheuring, S. & Rico, F. High-frequency microrheology reveals cytoskeleton dynamics in living cells. Nat. Phys. 13, 771–775 (2017).
Hassan, A. E. et al. Relative microelastic mapping of living cells by atomic force microscopy. Biophys. J. 74, 1564–1578 (1998).
Morse, D. C. Viscoelasticity of tightly entangled solutions of semiflexible polymers. Phys. Rev. E 58, R1237–R1240 (1998).
Broedersz, C. P. & MacKintosh, F. C. Modeling semiflexible polymer networks. Rev. Mod. Phys. 86, 995–1036 (2014).
Dimitriadis, E. K., Horkay, F., Maresca, J., Kachar, B. & Chadwick, R. S. Determination of elastic moduli of thin layers of soft material using the atomic force microscope. Biophys. J. 82, 2798–2810 (2002). This paper introduces a helpful approach to reliably extract the mechanical properties of spread cells, as it remedies the insufficient treatment of their finite thickness within the Hertzian (and related) model.
Storm, C., Pastore, J. J., MacKintosh, F. C., Lubensky, T. C. & Janmey, P. A. Nonlinear elasticity in biological gels. Nature 435, 191–194 (2005).
Notbohm, J., Poon, B. & Ravichandran, G. Analysis of nanoindentation of soft materials with an atomic force microscope. J. Mater. Res. 27, 229–237 (2012).
Friedrichs, J. et al. A practical guide to quantify cell adhesion using single-cell force spectroscopy. Methods 60, 169–178 (2013).
Hinterdorfer, P. & Dufrene, Y. F. Detection and localization of single molecular recognition events using atomic force microscopy. Nat. Methods 3, 347–355 (2006).
Snijder, J., Ivanovska, I. L., Baclayon, M., Roos, W. H. & Wuite, G. J. Probing the impact of loading rate on the mechanical properties of viral nanoparticles. Micron 43, 1343–1350 (2012).
Medalsy, I. D. & Muller, D. J. Nanomechanical properties of proteins and membranes depend on loading rate and electrostatic interactions. ACS Nano 7, 2642–2650 (2013).
Efremov, Y. M., Wang, W. H., Hardy, S. D., Geahlen, R. L. & Raman, A. Measuring nanoscale viscoelastic parameters of cells directly from AFM force-displacement curves. Sci. Rep. 7, 1541 (2017). This paper introduces a powerful approach to extracting elastic and viscous parameters from FD curves.
Zink, M. & Grubmuller, H. Mechanical properties of the icosahedral shell of southern bean mosaic virus: a molecular dynamics study. Biophys. J. 96, 1350–1363 (2009).
Schoeler, C. et al. Mapping mechanical force propagation through biomolecular complexes. Nano Lett. 15, 7370–7376 (2015).
Chaudhuri, O., Parekh, S. H., Lam, W. A. & Fletcher, D. A. Combined atomic force microscopy and side-view optical imaging for mechanical studies of cells. Nat. Methods 6, 383–387 (2009).
Beicker, K., O’Brien, E. T. 3rd, Falvo, M. R. & Superfine, R. Vertical light sheet enhanced side-view imaging for AFM cell mechanics studies. Sci. Rep. 8, 1504 (2018).
Curry, N., Ghezali, G., Kaminski Schierle, G. S., Rouach, N. & Kaminski, C. F. Correlative STED and atomic force microscopy on live astrocytes reveals plasticity of cytoskeletal structure and membrane physical properties during polarized migration. Front. Cell Neurosci. 11, 104 (2017).
Freikamp, A., Cost, A. L. & Grashoff, C. The piconewton force awakens: quantifying mechanics in cells. Trends Cell Biol. 26, 838–847 (2016).
Polacheck, W. J. & Chen, C. S. Measuring cell-generated forces: a guide to the available tools. Nat. Methods 13, 415–423 (2016).
Zou, P. et al. Bright and fast multicoloured voltage reporters via electrochromic FRET. Nat. Commun. 5, 4625 (2014).
Nekimken, A. L. et al. Pneumatic stimulation of C. elegans mechanoreceptor neurons in a microfluidic trap. Lab. Chip 17, 1116–1127 (2017).
Sakaue-Sawano, A. et al. Visualizing spatiotemporal dynamics of multicellular cell-cycle progression. Cell 132, 487–498 (2008).
Grashoff, C. et al. Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466, 263–266 (2010).
Guo, J., Wang, Y., Sachs, F. & Meng, F. Actin stress in cell reprogramming. Proc. Natl Acad. Sci. USA 111, E5252–E5261 (2014).
Freikamp, A., Mehlich, A., Klingner, C. & Grashoff, C. Investigating piconewton forces in cells by FRET-based molecular force microscopy. J. Struct. Biol. 197, 37–42 (2017).
Krieg, M., Dunn, A. R. & Goodman, M. B. Mechanical control of the sense of touch by beta-spectrin. Nat. Cell Biol. 16, 224–233 (2014).
O’Callaghan, R., Job, K. M., Dull, R. O. & Hlady, V. Stiffness and heterogeneity of the pulmonary endothelial glycocalyx measured by atomic force microscopy. Am. J. Physiol. Lung Cell. Mol. Physiol. 301, L353–L360 (2011).
Morris, C. E. & Homann, U. Cell surface area regulation and membrane tension. J. Membr. Biol. 179, 79–102 (2001).
Cartagena-Rivera, A. X., Logue, J. S., Waterman, C. M. & Chadwick, R. S. Actomyosin cortical mechanical properties in nonadherent cells determined by atomic force microscopy. Biophys. J. 110, 2528–2539 (2016).
Janmey, P. A. & Weitz, D. A. Dealing with mechanics: mechanisms of force transduction in cells. Trends Biochem. Sci. 29, 364–370 (2004).
Laurent, V. M. et al. Partitioning of cortical and deep cytoskeleton responses from transient magnetic bead twisting. Ann. Biomed. Eng. 31, 1263–1278 (2003).
Lammerding, J. Mechanics of the nucleus. Compr. Physiol. 1, 783–807 (2011).
Discher, D. E., Janmey, P. & Wang, Y. L. Tissue cells feel and respond to the stiffness of their substrate. Science 310, 1139–1143 (2005).
Engler, A. J., Sen, S., Sweeney, H. L. & Discher, D. E. Matrix elasticity directs stem cell lineage specification. Cell 126, 677–689 (2006).
Engler, A. J., Rehfeldt, F., Sen, S. & Discher, D. E. Microtissue elasticity: measurements by atomic force microscopy and its influence on cell differentiation. Methods Cell Biol. 83, 521–545 (2007).
Koser, D. E. et al. Mechanosensing is critical for axon growth in the developing brain. Nat. Neurosci. 19, 1592–1598 (2016).
Moeendarbary, E. et al. The soft mechanical signature of glial scars in the central nervous system. Nat. Commun. 8, 14787 (2017).
Hardie, R. C. & Franze, K. Photomechanical responses in Drosophila photoreceptors. Science 338, 260–263 (2012).
Peaucelle, A. et al. Pectin-induced changes in cell wall mechanics underlie organ initiation in Arabidopsis. Curr. Biol. 21, 1720–1726 (2011).
Zhang, T., Vavylonis, D., Durachko, D. M. & Cosgrove, D. J. Nanoscale movements of cellulose microfibrils in primary cell walls. Nat. Plants 3, 17056 (2017).
Lopez, J. I., Kang, I., You, W. K., McDonald, D. M. & Weaver, V. M. In situ force mapping of mammary gland transformation. Integr. Biol. 3, 910–921 (2011).
Plodinec, M. et al. The nanomechanical signature of breast cancer. Nat. Nanotechnol 7, 757–765 (2012).
Cross, S. E., Jin, Y. S., Rao, J. & Gimzewski, J. K. Nanomechanical analysis of cells from cancer patients. Nat. Nanotechnol. 2, 780–783 (2007). This work introduces AFM for the mechanical phenotyping of cancer cells derived from patients.
Iyer, S., Gaikwad, R. M., Subba-Rao, V., Woodworth, C. D. & Sokolov, I. Atomic force microscopy detects differences in the surface brush of normal and cancerous cells. Nat. Nanotechnol. 4, 389–393 (2009).
Staunton, J. R., Doss, B. L., Lindsay, S. & Ros, R. Correlating confocal microscopy and atomic force indentation reveals metastatic cancer cells stiffen during invasion into collagen I matrices. Sci. Rep. 6, 19686 (2016).
Barriga, E. H., Franze, K., Charras, G. & Mayor, R. Tissue stiffening coordinates morphogenesis by triggering collective cell migration in vivo. Nature 554, 523–527 (2018).
Krieg, M. et al. Genetic defects in beta-spectrin and tau sensitize C.elegans axons to movement-induced damage via torque-tension coupling. eLife 6, e20172 (2017).
Zhang, Y. et al. Modeling of the axon membrane skeleton structure and implications for its mechanical properties. PLOS Comput. Biol. 13, e1005407 (2017).
Magdesian, M. H. et al. Atomic force microscopy reveals important differences in axonal resistance to injury. Biophys. J. 103, 405–414 (2012).
Gueta, R., Barlam, D., Shneck, R. Z. & Rousso, I. Measurement of the mechanical properties of isolated tectorial membrane using atomic force microscopy. Proc. Natl Acad. Sci. USA 103, 14790–14795 (2006).
Szarama, K. B., Gavara, N., Petralia, R. S., Kelley, M. W. & Chadwick, R. S. Cytoskeletal changes in actin and microtubules underlie the developing surface mechanical properties of sensory and supporting cells in the mouse cochlea. Development 139, 2187–2197 (2012).
Windmill, J. F. C., Jackson, J. C., Pook, V. G. & Robert, D. Frequency doubling by active in vivo motility of mechanosensory neurons in the mosquito ear. R. Soc. Open Sci. 5, 171082 (2018).
Rotsch, C., Braet, F., Wisse, E. & Radmacher, M. AFM imaging and elasticity measurements on living rat liver macrophages. Cell Biol. Int. 21, 685–696 (1997).
Rotsch, C., Jacobson, K. & Radmacher, M. Dimensional and mechanical dynamics of active and stable edges in motile fibroblasts investigated by using atomic force microscopy. Proc. Natl Acad. Sci. USA 96, 921–926 (1999).
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).
Blaue, C., Kashef, J. & Franz, C. M. Cadherin-11 promotes neural crest cell spreading by reducing intracellular tension-Mapping adhesion and mechanics in neural crest explants by atomic force microscopy. Semin. Cell Dev. Biol. 73, 95–106 (2018).
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).
Matzke, R., Jacobson, K. & Radmacher, M. Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nat. Cell Biol. 3, 607–610 (2001). This is a pioneering AFM study mapping the dynamic stiffness changes of the cortex of adherent cells.
Gaub, B. M. & Muller, D. J. Mechanical stimulation of piezo1 receptors depends on extracellular matrix proteins and directionality of force. Nano Lett. 17, 2064–2072 (2017).
Ossola, D. et al. Force-controlled patch clamp of beating cardiac cells. Nano Lett. 15, 1743–1750 (2015).
Pamir, E., George, M., Fertig, N. & Benoit, M. Planar patch-clamp force microscopy on living cells. Ultramicroscopy 108, 552–557 (2008).
Upadhye, K. V., Candiello, J. E., Davidson, L. A. & Lin, H. Whole-cell electrical activity under direct mechanical stimulus by AFM cantilever using planar patch clamp chip approach. Cell. Mol. Bioeng. 4, 270–280 (2011).
Martinez-Martin, D. et al. Inertial picobalance reveals fast mass fluctuations in mammalian cells. Nature 550, 500–505 (2017). This paper introduces AFM as a picobalance to measure the total mass of adherent cells at millisecond time resolution under cell culture conditions.
Xu, W. et al. Modeling and measuring the elastic properties of an archaeal surface, the sheath of Methanospirillum hungatei, and the implication for methane production. J. Bacteriol. 178, 3106–3112 (1996).
Yao, X., Jericho, M., Pink, D. & Beveridge, T. Thickness and elasticity of gram-negative murein sacculi measured by atomic force microscopy. J. Bacteriol. 181, 6865–6875 (1999).
Gaboriaud, F., Bailet, S., Dague, E. & Jorand, F. Surface structure and nanomechanical properties of Shewanella putrefaciens bacteria at two pH values (4 and 10) determined by atomic force microscopy. J. Bacteriol. 187, 3864–3868 (2005).
van der Mei, H. C. et al. Direct probing by atomic force microscopy of the cell surface softness of a fibrillated and nonfibrillated oral streptococcal strain. Biophys. J. 78, 2668–2674 (2000).
Francius, G., Domenech, O., Mingeot-Leclercq, M. P. & Dufrene, Y. F. Direct observation of Staphylococcus aureus cell wall digestion by lysostaphin. J. Bacteriol. 190, 7904–7909 (2008).
Touhami, A., Nysten, B. & Dufrêne, Y. F. Nanoscale mapping of the elasticity of microbial cells by atomic force microscopy. Langmuir 19, 4539–4543 (2003).
Francius, G., Tesson, B., Dague, E., Martin-Jezequel, V. & Dufrene, Y. F. Nanostructure and nanomechanics of live Phaeodactylum tricornutum morphotypes. Environ. Microbiol. 10, 1344–1356 (2008).
Mosier, A. P., Kaloyeros, A. E. & Cady, N. C. A novel microfluidic device for the in situ optical and mechanical analysis of bacterial biofilms. J. Microbiol. Methods 91, 198–204 (2012).
Alsteens, D., Trabelsi, H., Soumillion, P. & Dufrene, Y. F. Multiparametric atomic force microscopy imaging of single bacteriophages extruding from living bacteria. Nat. Commun. 4, 2926 (2013).
Formosa-Dague, C., Speziale, P., Foster, T. J., Geoghegan, J. A. & Dufrene, Y. F. Zinc-dependent mechanical properties of Staphylococcus aureus biofilm-forming surface protein SasG. Proc. Natl Acad. Sci. USA 113, 410–415 (2016).
Marchetti, M., Wuite, G. & Roos, W. H. Atomic force microscopy observation and characterization of single virions and virus-like particles by nano-indentation. Curr. Opin. Virol. 18, 82–88 (2016).
Ivanovska, I. L. et al. Bacteriophage capsids: tough nanoshells with complex elastic properties. Proc. Natl Acad. Sci. USA 101, 7600–7605 (2004).
Uetrecht, C. et al. High-resolution mass spectrometry of viral assemblies: molecular composition and stability of dimorphic hepatitis B virus capsids. Proc. Natl Acad. Sci. USA 105, 9216–9220 (2008).
Smith, D. E. et al. The bacteriophage straight phi29 portal motor can package DNA against a large internal force. Nature 413, 748–752 (2001).
Carrasco, C. et al. DNA-mediated anisotropic mechanical reinforcement of a virus. Proc. Natl Acad. Sci. USA 103, 13706–13711 (2006). This paper shows that the viral genome interacts with and mechanically reinforces the viral capsid.
Castellanos, M., Carrillo, P. J. & Mateu, M. G. Quantitatively probing propensity for structural transitions in engineered virus nanoparticles by single-molecule mechanical analysis. Nanoscale 7, 5654–5664 (2015).
Snijder, J. et al. Probing the biophysical interplay between a viral genome and its capsid. Nat. Chem. 5, 502–509 (2013).
Carrasco, C. et al. Built-in mechanical stress in viral shells. Biophys. J. 100, 1100–1108 (2011).
Baclayon, M. et al. Prestress strengthens the shell of Norwalk virus nanoparticles. Nano Lett. 11, 4865–4869 (2011). This work describes the contribution of viral domains to the mechanical stability of a viral capsid.
Roos, W. H. et al. Mechanics of bacteriophage maturation. Proc. Natl Acad. Sci. USA 109, 2342–2347 (2012).
Kol, N. et al. A stiffness switch in human immunodeficiency virus. Biophys. J. 92, 1777–1783 (2007). This work establishes a direct link between the mechanical properties of HIV and its infectivity.
Ortega-Esteban, A. et al. Fluorescence tracking of genome release during mechanical unpacking of single viruses. ACS Nano 9, 10571–10579 (2015).
Martinez-Martin, D. et al. Resolving structure and mechanical properties at the nanoscale of viruses with frequency modulation atomic force microscopy. PLOS ONE 7, e30204 (2012).
Luque, D. Self-assembly and characterization of small and monodisperse dye nanospheres in a protein cage. Chem. Sci. 5, 575–581 (2014).
Heinze, K. et al. Protein nanocontainers from nonviral origin: testing the mechanics of artificial and natural protein cages by AFM. J. Phys. Chem. B 120, 5945–5952 (2016).
Snijder, J. et al. Assembly and mechanical properties of the cargo-free and cargo-loaded bacterial nanocompartment encapsulin. Biomacromolecules 17, 2522–2529 (2016).
Llauro, A. et al. Decrease in pH destabilizes individual vault nanocages by weakening the inter-protein lateral interaction. Sci. Rep. 6, 34143 (2016).
Sieben, C. et al. Influenza virus binds its host cell using multiple dynamic interactions. Proc. Natl Acad. Sci. USA 109, 13626–13631 (2012).
Alsteens, D. et al. Nanomechanical mapping of first binding steps of a virus to animal cells. Nat. Nanotechnol. 12, 177–183 (2017).
Arkhipov, A., Roos, W. H., Wuite, G. J. & Schulten, K. Elucidating the mechanism behind irreversible deformation of viral capsids. Biophys. J. 97, 2061–2069 (2009).
Krieg, M. et al. Tensile forces govern germ-layer organization in zebrafish. Nat. Cell Biol. 10, 429–436 (2008). This work addresses a question in cell biology that is over 30 years old, that is, whether cell adhesion or cortex tension governs germ-layer organization in development.
Heu, C., Berquand, A., Elie-Caille, C. & Nicod, L. Glyphosate-induced stiffening of HaCaT keratinocytes, a peak force tapping study on living cells. J. Struct. Biol. 178, 1–7 (2012).
Sheetz, M. P. Cell control by membrane-cytoskeleton adhesion. Nat. Rev. Mol. Cell Biol. 2, 392–396 (2001).
Krieg, M., Helenius, J., Heisenberg, C. P. & Muller, D. J. A bond for a lifetime: employing membrane nanotubes from living cells to determine receptor-ligand kinetics. Angew. Chem. Int. Ed. 47, 9775–9777 (2008).
Vasquez, V., Krieg, M., Lockhead, D. & Goodman, M. B. Phospholipids that contain polyunsaturated fatty acids enhance neuronal cell mechanics and touch sensation. Cell Rep. 6, 70–80 (2014).
Gonnermann, C. et al. Quantitating membrane bleb stiffness using AFM force spectroscopy and an optical sideview setup. Integr. Biol. 7, 356–363 (2015).
Dong, M., Husale, S. & Sahin, O. Determination of protein structural flexibility by microsecond force spectroscopy. Nat. Nanotechnol. 4, 514–517 (2009).
Medalsy, I., Hensen, U. & Muller, D. J. Imaging and quantifying chemical and physical properties of native proteins at molecular resolution by force-volume AFM. Angew. Chem. Int. Ed. 50, 12103–12108 (2011).
Rico, F., Su, C. & Scheuring, S. Mechanical mapping of single membrane proteins at submolecular resolution. Nano Lett. 11, 3983–3986 (2011).
Pfreundschuh, M., Martinez-Martin, D., Mulvihill, E., Wegmann, S. & Muller, D. J. Multiparametric high-resolution imaging of native proteins by force-distance curve-based AFM. Nat. Protoc. 9, 1113–1130 (2014).
Liang, X., Mao, G. & Simon Ng, K. Y. Probing small unilamellar EggPC vesicles on mica surface by atomic force microscopy. Colloids Surf. B Biointerfaces 34, 41–51 (2004).
Delorme, N. & Fery, A. Direct method to study membrane rigidity of small vesicles based on atomic force microscope force spectroscopy. Phys. Rev. E Stat. Nonlin. Soft Matter Phys. 74, 030901 (2006).
Li, S., Eghiaian, F., Sieben, C., Herrmann, A. & Schaap, I. A. Bending and puncturing the influenza lipid envelope. Biophys. J. 100, 637–645 (2011).
Vorselen, D. et al. Multilamellar nanovesicles show distinct mechanical properties depending on their degree of lamellarity. Nanoscale 10, 5318–5324 (2018).
Engel, A. & Muller, D. J. Observing single biomolecules at work with the atomic force microscope. Nat. Struct. Biol. 7, 715–718 (2000).
Muller, D. J., Buldt, G. & Engel, A. Force-induced conformational change of bacteriorhodopsin. J. Mol. Biol. 249, 239–243 (1995).
Pfreundschuh, M., Hensen, U. & Muller, D. J. Quantitative imaging of the electrostatic field and potential generated by a transmembrane protein pore at subnanometer resolution. Nano Lett. 13, 5585–5593 (2013).
Martinez-Martin, D., Herruzo, E. T., Dietz, C., Gomez-Herrero, J. & Garcia, R. Noninvasive protein structural flexibility mapping by bimodal dynamic force microscopy. Phys. Rev. Lett. 106, 198101 (2011).
Wegmann, S. et al. Human Tau isoforms assemble into ribbon-like fibrils that display polymorphic structure and stability. J. Biol. Chem. 285, 27302–27313 (2010).
Zhang, S. et al. Coexistence of ribbon and helical fibrils originating from hIAPP(20–29) revealed by quantitative nanomechanical atomic force microscopy. Proc. Natl Acad. Sci. USA 110, 2798–2803 (2013).
de Pablo, P. J., Schaap, I. A. T., MacKintosh, F. C. & Schmidt, C. F. Deformation and collapse of microtubules on the nanometer scale. Phys. Rev. Lett. 91, 098101 (2003).
Block, J. et al. Nonlinear loading-rate-dependent force response of individual vimentin intermediate filaments to applied strain. Phys. Rev. Lett. 118, 048101 (2017).
Goodman, R. P. et al. Rapid chiral assembly of rigid DNA building blocks for molecular nanofabrication. Science 310, 1661–1665 (2005).
Radmacher, M., Tillmann, R. W., Fritz, M. & Gaub, H. E. From molecules to cells: imaging soft samples with the atomic force microscope. Science 257, 1900–1905 (1992).
Adamantidis, A. et al. Optogenetics: 10 years after ChR2 in neurons — views from the community. Nat. Neurosci. 18, 1202–1212 (2015).
Seo, D. et al. A mechanogenetic toolkit for interrogating cell signaling in space and time. Cell 169, 1357 (2017).
Dufrene, Y. F. et al. Five challenges to bringing single-molecule force spectroscopy into living cells. Nat. Methods 8, 123–127 (2011).
Norregaard, K., Metzler, R., Ritter, C. M., Berg-Sorensen, K. & Oddershede, L. B. Manipulation and motion of organelles and single molecules in living cells. Chem. Rev. 117, 4342–4375 (2017).
Derjaguin, B. V., Muller, V. M. & Toporov, Y. P. Effect of contact deformations on adhesion of particles. J. Colloid Interface Sci. 53, 314–326 (1975).
Lomakina, E. B., Spillmann, C. M., King, M. R. & Waugh, R. E. Rheological analysis and measurement of neutrophil indentation. Biophys. J. 87, 4246–4258 (2004).
Rosenbluth, M. J., Lam, W. A. & Fletcher, D. A. Force microscopy of nonadherent cells: a comparison of leukemia cell deformability. Biophys. J. 90, 2994–3003 (2006).
de Sousa, J. S. et al. Analytical model of atomic-force-microscopy force curves in viscoelastic materials exhibiting power law relaxation. J. Appl. Phys. 121, 034901 (2017).
Darling, E. M., Zauscher, S. & Guilak, F. Viscoelastic properties of zonal articular chondrocytes measured by atomic force microscopy. Osteoarthritis Cartilage 14, 571–579 (2006).
Stifter, T., Weilandt, E., Marti, O. & Hild, S. Influence of the topography on adhesion measured by SFM. Appl. Phys. A 66, S597–S605 (1998).
Rico, F. et al. Probing mechanical properties of living cells by atomic force microscopy with blunted pyramidal cantilever tips. Phys. Rev. E 72, 021914 (2005).
The authors thank R. Newton for critically discussing the manuscript. M.K. acknowledges financial support from the Spanish Ministry of Economy and Competitiveness through the Ramon y Cajal programme (RYC-2015-17935), “Severo Ochoa” programme for the Centres of Excellence in R&D (SEV-2015-0522) from Fundació Privada Cellex, Generalitat de Catalunya, through the Centres de Recerca de Catalunya (CERCA) programme, and the European Research Council (ERC; MechanoSystems grant 715243). D.A. was supported by the ERC (NanoVirus grant 758224) and the National Fund for Scientific Research and Research Department of the Communauté française de Belgique (Concerted Research Action). B.M.G. was supported by a long-term European Molecular Biology Organization (EMBO) fellowship (ALTF 424–2016). W.H.R. was funded by the Nederlandse organisatie voor Wetenschappelijk Onderzoek (VIDI grant). H.E.G. acknowledges financial support from the CelluFuel ERC grant. C.G. was supported by the Swiss Nanoscience Institute (SNI), University of Basel. Y.F.D. was supported by the Université catholique de Louvain, ERC, under the European Union’s Horizon 2020 research and innovation programme (grant 693630), Walloon Excellence in Life Sciences and Biotechnology (WELBIO) (grant no. WELBIO-CR-2015A-05), National Fund for Scientific Research (FNRS and EOS grants) and Research Department of the Communauté française de Belgique (Concerted Research Action). D.J.M. was supported by the Swiss National Science Foundation (SNF; grant 310030B_160225), the National Centre of Competence in Research (NCCR) Molecular Systems Engineering and the Swiss Commission for Technology and Innovation (CTI, grant 28033.1).
The authors declare no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Krieg, M., Fläschner, G., Alsteens, D. et al. Atomic force microscopy-based mechanobiology. Nat Rev Phys 1, 41–57 (2019). https://doi.org/10.1038/s42254-018-0001-7
- Biological Sensor Systems
- Mechanical Cues
- Actomyosin Cortex
- Atomic Force Microscopy Probe
- Cortical Shell-liquid Core (CSLC)
This article is cited by
Trends in mechanobiology guided tissue engineering and tools to study cell-substrate interactions: a brief review
Biomaterials Research (2023)
High-resolution line-scan Brillouin microscopy for live imaging of mechanical properties during embryo development
Nature Methods (2023)
High-force catch bonds between the Staphylococcus aureus surface protein SdrE and complement regulator factor H drive immune evasion
Communications Biology (2023)
Does “form follow function” in the rotiferan genus Keratella?
Mechanical stimulation devices for mechanobiology studies: a market, literature, and patents review
Bio-Design and Manufacturing (2023)