Abstract
A single elastic modulus is not sufficient for describing the mechanical behavior of a living cell due to its viscoelastic nature and heterogeneity beneath the membrane. In this paper, the nanoscale elastic and viscoelastic behavior of individual living Chinese hamster ovary (CHO-K1) cells in a physiological environment were probed by atomic force microscopy (AFM) indentations at various loading rates. Based on Hertzian fits of the force–distance curves, the apparent elastic modulus of the cells was determined and found to be a function of the loading rate as well as the indentation depth. Notably, contributions from the substrate were negligible up to 50% of the cell thickness. For increased indentation rates and depths, healthy spindle-shaped CHO-K1 cells were found to exhibit an increased change of stiffness, whereas for unhealthy oval- shaped CHO-K1 cells there was little stiffening at equivalent loading rates and depths. Furthermore, a larger hysteresis between the loading and unloading curves was observed with increasing loading rates, which was related to the viscoelastic behavior of CHO-K1 cells. This work demonstrates differences in the rate- and depth-dependent elastic behavior at the nanoscale level between healthy and unhealthy mammalian cells.
Similar content being viewed by others
References
M. Radmacher, R.W. Tillmann, M. Fritz, H.E. Gaub: From molecules to cells: Imaging soft samples with the atomic force microscope. Science 257, 1900 (1992).
H. Huang, R. Kamm, R. Lee: Cell mechanics and mechanotransduction: Pathways, probes, and physiology. Am. J. Physiol. 287, 1 (2004).
G. Bao, S. Suresh: Cell and molecular mechanics of biological materials. Nat. Mater. 2, 715 (2003).
G. Charras, M. Horton: Single cell mechanotransduction and its modulation analyzed by atomic force microscope indentation. Biophys. J. 82, 2970 (2002).
A. Ikai, R. Afrin: Toward mechanical manipulations of cell membranes and membrane proteins using an atomic force microscope: An invited review. Cell Biochem. Biophys. 39, 257 (2003).
K. Vliet, G. Bao, S. Suresh: The biomechanics toolbox: Experimental approaches for living cells and biomolecules. Acta Mater. 51, 5881 (2003).
C. Zhu, G. Bao, N. Wang: Cell mechanics: Mechanical response, cell adhesion, and molecular deformation. Ann. Rev. Biomed. Eng. 2, 189 (2000).
S. Kasas, X. Wang, H. Hirling, R. Marsault, B. Huni, A. Yersin, R. Regazzi, G. Grenningloh, B. Riederer: Superfical and deep changes of celluar mechanical properties following cytoskeleton disassembly. Cell Motil. Cytoskeleton 62, 124 (2005).
F. Braet, C. Rotsch, E. Wisse, M. Radmacher: Comparison of fixed and living liver endothelial cells by atomic force microscopy. Appl. Phys. A 66, S (1998).
E. Takai, K. Costa, A. Shaheen, C. Hung, X. Guo: Osteoblast elastic modulus measured by atomic force microscopy is substrate dependent. Ann. Biomed. Eng. 33, 963 (2005).
C. Rotsch, K. Jacobson, M. Radmacher: 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 (1999).
H. Haga, S. Sasaki, K. Kawabata, E. Ito, T. Ushiki, T. Sambongi: Elasticity mapping of living fibroblasts by AFM and immunofluorescence observation of the cytoskeleton. Ultramicroscopy 82, 253 (2000).
A. Mathur, G. Truskey, W. Reichert: Atomic force and total internal reflection fluorescence microscopy for the study of force transmission in endothelial cells. Biophys. J. 78, 1725 (2000).
R. Matzke, K. Jacobson, M. Radmacher: Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nat. Cell Biol. 3, 607 (2001).
P. Haydon, R. Lartius, V. Parpura, S. Marchese-Ragona: Membrane deformation of living glial cells using atomic force microscopy. J. Microsc. 182, 114 (1996).
I. Lee, R. Marchant: Force measurements on platelet surfaces with high spatial resolution under physiological conditions. Colloids Surf. B Biointerfaces 19, 357 (2000).
C. Rotsch, M. Radmacher: Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: An atomic force microscopy study. Biophys. J. 78, 520 (2000).
M. McElfresh, E. Baesu, R. Balhorn, J. Belak, M.J. Allen, R.E. Rudd: Combining constitutive materials modeling with atomic force microscopy to understand the mechanical properties of living cells. Proc. Natl. Acad. Sci. USA 99, 6493 (2002).
J. Hutter, J. Chen, W.K. Wan, S. Uniyal, M. Leabu, B.M.C Chan: Atomic force microscopy investigation of the dependence of cellular elastic moduli on glutaraldehyde fixation. J. Microsc. 219, 61 (2005).
A. Pelling, Y. Li, W. Shi, J. Gimzewski: Nanoscale visualization and characterization of Myxococcus xanthus cells with atomic force microscopy. Proc. Natl. Acad. Sci. USA 102, 6484 (2005).
E. A-Hassan, W.F. Heinz, M.D. Antonik, N.P. D’Costa, S. Nageswaran, C.A. Schoenberger, J.H. Hoh: Relative microelastic mapping of living cells by atomic force microscopy. Biophys. J. 74, 1564 (1998).
S. Sen, S. Subramanian, D. Discher: Indentation and adhesive probing a cell membrane with AFM: Theoretical model and experiments. Biophys. J. 89, 3203 (2005).
K.S. Tai, H.J. Qi, C. Ortiz: Effect of mineral content on the nanoindentation properties and nanoscale deformation mechanisms of bovine tibial cortical bone. J. Mater. Sci.: Mater. Med. 16, 947 (2005).
A. Hategan, R. Law, S. Kahn, D. Discher: Adhesively tensed cell membranes: Lysis kinetics and atomic force microscopy probing. Biophys. J. 85, 2746 (2003).
S. Heidemann, D. Wirtz: Towards a regional approach to cell mechanics. Trends Cell Biol. 14, 160 (2004).
H. Wu, V. Moy: Mechanical properties of L929 cells measured by atomic force microscopy: Effects of anticytoskeletal drugs and membrane crosslinking. Scanning 20, 389 (1998).
M. Beil, A. Micoulet, G.V. Wichert, S. Paschke, P. Walther, M.B. Omary, P.P.V Veldhoven, U. Gern, E. Wolf-Hieber, J. Eggermann, J. Waltenberger, G. Adler, J. Spatz, T. Seufferlein: Sphingosylphosphorylcholine regulates keratin network architecture and visco-elastic properties of human cancer cells. Nat. Cell Biol. 5, 803 (2003).
S. Yamada, D. Wirtz, S.C. Kuo: Mechanics of living cells measured by laser tracking microrheology. Biophys. J. 78, 1736 (2000).
A. Mathur, A. Collinsworth, W. Reichert, W. Kraus, G. Truskey: Endothelial, cardiac muscle and skeletal muscle exhibit different viscous and elastic properties as determined by atomic force microscopy. J. Biomech. 34, 1545 (2001).
F. Rico, P. Roca-Cusachs, N. Gavara, R. Farre, M. Rotger, D. Navajas: Probing mechanical properties of living cells by atomic force microscopy with blunted pyramidal cantilever tips. Phys. Rev. E 72, 021914 (2005).
J. Alcaraz, L. Buscemi, M. Grabulosa, X. Trepat, B. Fabry, R. Farre, D. Navajas: Microrheology of human lung epithelial cells measured by atomic force microscopy. Biophys. J. 84, 2071 (2003).
T. Tsui, G. Pharr: Substrate effects on nanoindentation mechanical property measurement of soft films on hard substrate. J. Mater. Res. 14, 292 (1999).
J. Domke, M. Radmacher: Measuring the elastic properties of thin polymer films with the atomic force microscope. Langmuir 14, 3320 (1998).
T. Berdyyeva, C. Woodworth, I. Sokolov: Human epithelial cells increase their rigidity with ageing in vitro: Direct measurements. Phys. Med. Biol. 50, 81 (2005).
K. Costa: Single-cell elastography: Probing for disease with the atomic force microscope. Dis. Markers 19, 139 (2003).
S. Suresh, J. Spatz, J.P. Mills, A. Micoulet, M. Dao, C.T. Lim, M. Beil, T. Seufferlein: Connections between single-cell biomechanics and human disease states: Gastrointestinal cancer and malaria. Acta Biomater. 1, 15 (2005).
E. Stoffels, I. Kieft, R. Sladek: Superfical treatment of mammalian cells using plasma needle. J. Phys. D: Appl. Phys. 36, 2908 (2003).
K. Costa, F. Yin: Analysis of indentation: Implications for measuring mechical properties with atomic force microscopy. J. Biomech, Eng. Trans. ASME 121, 462 (1999).
E. Dimitriadis, F. Horkay, J. Maresca, B. Kachar, R. Chadwick: Determination of elastic moduli of thin layers of soft material using the atomic force microscope. Biophys. J. 82, 2798 (2002).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, M., Srinivasan, C., Burgess, D.J. et al. Rate- and depth-dependent nanomechanical behavior of individual living Chinese hamster ovary cells probed by atomic force microscopy. Journal of Materials Research 21, 1906–1912 (2006). https://doi.org/10.1557/jmr.2006.0233
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1557/jmr.2006.0233