European Biophysics Journal

, Volume 37, Issue 6, pp 935–945 | Cite as

Probing elasticity and adhesion of live cells by atomic force microscopy indentation

  • L. SirghiEmail author
  • J. Ponti
  • F. Broggi
  • F. RossiEmail author
Original Paper


Atomic force microscopy (AFM) indentation has become an important technique for quantifying the mechanical properties of live cells at nanoscale. However, determination of cell elasticity modulus from the force–displacement curves measured in the AFM indentations is not a trivial task. The present work shows that these force–displacement curves are affected by indenter-cell adhesion force, while the use of an appropriate indentation model may provide information on the cell elasticity and the work of adhesion of the cell membrane to the surface of the AFM probes. A recently proposed indentation model (Sirghi, Rossi in Appl Phys Lett 89:243118, 2006), which accounts for the effect of the adhesion force in nanoscale indentation, is applied to the AFM indentation experiments performed on live cells with pyramidal indenters. The model considers that the indentation force equilibrates the elastic force of the cell cytoskeleton and the adhesion force of the cell membrane. It is assumed that the indenter-cell contact area and the adhesion force decrease continuously during the unloading part of the indentation (peeling model). Force–displacement curves measured in indentation experiments performed with silicon nitride AFM probes with pyramidal tips on live cells (mouse fibroblast Balb/c3T3 clone A31-1-1) in physiological medium at 37°C agree well with the theoretical prediction and are used to determine the cell elasticity modulus and indenter-cell work of adhesion.


Cell mechanics Atomic force microscopy indentation Cell membrane adhesion 



We are grateful to Mr. Takao Sasaki for SEM images of the AFM probes used in the experiments.

Supplementary material

249_2008_311_MOESM1_ESM.doc (674 kb)
Electronic supplementary material (DOC 674 kb)


  1. Afrin R, Yamada T, Ikai A (2004) Analysis of force curves obtained on the live cell membrane using chemically modified AFM probes. Ultramicroscopy 100:187–195CrossRefGoogle Scholar
  2. A-Hassan E, Heinz WF, Antonik MD, D’Costa NP, Nageswaran S, Schoenenberger C-A, Hoh JH (1998) Relative microelastic mapping of living cells by atomic force microscopy. Biophys J 74:1564–1578Google Scholar
  3. Alcaraz J, Buscemi L, Grabulosa M, Trepat X, Fabry B, Farree R, Navajas D (2003) Microrheology of human lung epithelial cells measured by atomic force microscopy. Biophys J 84:2071–2079Google Scholar
  4. Andersen L K, Cortera SA, Justesen J, Duch M, Hansen O, Chevallier J, Foss M, Pedersen FS and, Besenbacher F (2005) Cell volume increase in murine MC3T3-E1 Pre-ostereoblasts attaching onto biocompatible tantalum observed by magnetic AC mode atomic force microscopy. Euro Cells Mater 10:61–69Google Scholar
  5. Antunes JM, Menezes LF, Fernandes JV (2006) Three-dimensional numerical simulation of Vickers indentation tests. Int J Solids Struct 43:784–806zbMATHCrossRefGoogle Scholar
  6. Burnham NA, Chen X, Hodges CS, Matei GA, Thoreson EJ, Roberts CJ, Davies MC, Tendler SJB (2003) Comparison of calibration methods for atomic-force microscopy cantilevers. Nanotechnology 14:1–6CrossRefADSGoogle Scholar
  7. Indrajit R, Tymish YO, Dhruba JB, Haridas EP, Ruth AM, Navjot K, Paras NP (2005) Optical tracking of organically modified silica nanoparticles as DNA carriers: a nonviral nanomedicine approach for gene delivery. PNAS 102:279–284CrossRefGoogle Scholar
  8. Israelachivili JN (1992) Intermolecular and surface forces, 2nd edn. Academic Press, LondonGoogle Scholar
  9. Jena BP (2002) Fusion pore in live cells. News Physiol Sci 17:219–222Google Scholar
  10. Johnson KL, Kendall K, Roberts AD (1971) Surface energy and the contact of elastic solids. Proc R Soc Lond A324:301–313ADSGoogle Scholar
  11. Jung Y-G, Lawn BR, Martyniuk M, Huang H, Hu XZ (2004) Evaluation of elastic modulus and hardness of thin films by nanoindentation. J Mater Res 19:3076–3080CrossRefADSGoogle Scholar
  12. King RB (1987) Elastic analysis of some punch problems for a layered medium. Int J Solids Struct 23:1657–1664zbMATHCrossRefGoogle Scholar
  13. Lim CT, Zhou EH, Quek ST (2006) Mechanical models for living cells-a review. J Biomech 39:195–216CrossRefGoogle Scholar
  14. Murphy MF, Lalor MJ, Manning FCR, Lilley F, Crosby SR, Randall C and, Burton DR (2006) Comparative study of the conditions required to image live human epithelial and fibroblast cells using atomic force microscopy. Microsc Res Tech 69:757–765CrossRefGoogle Scholar
  15. McNamee CE, Nayoung P, Tanaka S, Kanda Y and, Higashitani K (2006a) Imaging of a soft, weakly adsorbing, living cell with a colloid probe tapping atomic force microscope technique. Colloids Surf B Biointerf 47:85–89CrossRefGoogle Scholar
  16. McNamee CE, Pyo N, Tanaka S, Vakarelski IU, Kanda Y, Higashitani K (2006b) Parameters affecting the adhesion strength between a living cell and a colloid probe when measured by the atomic force microscope. Colloids Surf B Biointerf 48:176–182CrossRefGoogle Scholar
  17. Obataya I, Nakamura Ch, Han SW, Nakamura N, Miyake J (2005) Nanoscale operation of a living cell using an atomic force microscope with nanoneedle. Nano Lett 5:27–30CrossRefADSGoogle Scholar
  18. Oberdorster G, Oberdorster EJ (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Perspect 113:823–839CrossRefGoogle Scholar
  19. Oliver WC, Pharr GM (1992) An improved technique for determining hardness and elastic modulus using load and displacement sensing indentation experiments. J Mater Res 7:1564–1583CrossRefADSGoogle Scholar
  20. Pesen D, Hoh JH (2005) Micromechanical architecture of the endothelial cell cortex. Biophys J 88:670–679CrossRefGoogle Scholar
  21. Rabinovich Y, Esayanur M, Daosukho S, Byer K, El-Shall H, Khan S (2005) Atomic force microscopy measurement of the elastic properties of the kidney epithelial cells. J Colloid Interface Sci 285:125–135CrossRefGoogle Scholar
  22. Robert A, Freitas Jr JD (2005) What is nanomedicine? Nanomed Nanotechnol Biol Med 1:2–9CrossRefGoogle Scholar
  23. Rotsch C, Radmacher M (2000) Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study. Biophys J 78:520–535Google Scholar
  24. Radmacher M (2002) Measuring the elastic properties of living cells by the atomic force microscopy. In: Jena BP, Horber JK (eds) Methods in Cell Biology, vol 68. Academic Press, Elsevier, New York, Amsterdam, pp 67–90Google Scholar
  25. Rotsch C, Braet F, Wisse E, Radmacher M (1997) AFM imaging and elasticity measurements on living rat liver macrophages. Cell Biol Int 11:685–696CrossRefGoogle Scholar
  26. Schaus SS, Henderson ER (1997) Cell viability and probe-cell membrane interactions of XR1 glial cells imaged by atomic force microscopy. Biophys J 73:1205–1214Google Scholar
  27. Seifert U, Lipowsky R (1995) The structure and dynamics of membranes. In: Lipowsky R, Sackmann E (eds) Handbook of biological physics, vol 1. Elsevier, AmsterdamGoogle Scholar
  28. Sen S, Subramanian S, Disher DE (2005) Indentation and adhesive probing of cell memmbrane with AFM: theoretical model and experiments. Biophys J 89:3203–3213CrossRefGoogle Scholar
  29. Simon A, Durrieu M-C (2006) Review. Strategies and results of atomic force microscopy in the study of cellular adhesion. Micron 37:1–13CrossRefGoogle Scholar
  30. Shen Y, Sun J L, Zhang A, Hu J, Xu L X (2007) A new image correction method for live cell atomic force microscopy. Phys Med Biol 52:2185–2196CrossRefGoogle Scholar
  31. Sirghi L, Rossi F (2006) Adhesion and elasticity in nanoscale indentation. Appl Phys Lett 89:243118–243120CrossRefADSGoogle Scholar
  32. Sneddon I N (1965) The relation between load and penetration in the axisymmetric Boussinesq problem for a punch of arbitrary profile. Int J Eng Sci 3:47–57zbMATHCrossRefMathSciNetGoogle Scholar
  33. Sun SX, Wirtz D (2006) Mechanics of enveloped virus entry into host cells. Biophys J 89:L10–12CrossRefGoogle Scholar
  34. Sun M, Graham J S, Hagedus B, Marga F, Zhang Y, Forgacs G, Grandbois M (2005) Multiple membrane tethers probed by atomic force microscopy. Biophys J 89:4320–4329CrossRefGoogle Scholar
  35. Zhu Ch (2000) Kinetick and mechanics of cell adhesion. J Biomech 33:23–33CrossRefGoogle Scholar
  36. Zhu AP, Fang N, Chan-Park MB, Chan V (2006) Adhesion contact dynamics of 3T3 fibroblasts on poly (lactide-co-glycoide acid). Surf Modified Photochem Immobil Biomacromolec Biomater 27:2566–2576Google Scholar

Copyright information

© EBSA 2008

Authors and Affiliations

  1. 1.European Commission, Joint Research CenterInstitute for Health and Consumer ProtectionIspra (VA)Italy

Personalised recommendations