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Nanomechanical Investigation of Soft Biological Cell Adhesion using Atomic Force Microscopy

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Abstract

Mechanical coupling between living cells is a complex process that is important for a variety of biological processes. In this study the effects of specific biochemical treatment on cell-to-cell adhesion and single cell mechanics were systematically investigated using atomic force microscopy (AFM) single cell force spectroscopy. Functionalised AFM tipless cantilevers were used for attaching single suspended cells that were brought in contact with substrate cells. Cell-to-cell adhesion parameters, such as maximum unbinding force (F max) and work or energy of detachment (W D), were extracted from the retraction force–displacement (F–d) curves. AFM indentation experiments were performed by indenting single cells with a spherical microbead attached to the cantilever. Hertzian contact model was applied to determine the elastic modulus (E) of single cells. Following treatment of the cells with neutralising antibody for epithelial (E)-cadherin, F max was increased by 25%, whereas W D decreased by 11% in response to a 43% increase in E. The results suggest that although the adhesion force between cells was increased after treatment, the energy of adhesion was decreased due to the reduced displacement separation as manifested by the loss of elastic deformation. Conclusively, changes in single cell mechanics are important underlying factors contributing to cell-to-cell adhesion and hence cytomechanical characterization is critical for cell adhesion measurements.

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Acknowledgments

This work was supported by grants from Diabetes UK (BDA: 09/0003913, and 12/0004546) and Leverhulme Trust (PRG-2012-738). The authors are grateful to JPK instruments for their technical support in the use of single-cell force spectroscopy.

Conflict of interest

The authors, Eleftherios Siamantouras, Claire E. Hills, Paul E. Squires and Kuo-Kang Liu, declare that they have no conflict of interests.

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No human nor animal studies were carried out by the authors for this article.

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Authors and Affiliations

  1. School of Engineering, University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, UK

    Eleftherios Siamantouras & Kuo-Kang Liu

  2. School of Life Sciences, University of Lincoln, Brayford Pool, Lincoln, LN6 7TS, UK

    Claire E. Hills & Paul E. Squires

Authors
  1. Eleftherios Siamantouras
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  2. Claire E. Hills
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  3. Paul E. Squires
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  4. Kuo-Kang Liu
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Correspondence to Kuo-Kang Liu.

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Associate Editor Cheng Dong oversaw the review of this article.

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Siamantouras, E., Hills, C.E., Squires, P.E. et al. Nanomechanical Investigation of Soft Biological Cell Adhesion using Atomic Force Microscopy. Cel. Mol. Bioeng. 8, 22–31 (2015). https://doi.org/10.1007/s12195-014-0359-2

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  • DOI: https://doi.org/10.1007/s12195-014-0359-2

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