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Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy

Nature Nanotechnology volume 6, pages 809–814 (2011)Cite this article

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Abstract

The nanomechanical properties of living cells, such as their surface elastic response and adhesion, have important roles in cellular processes such as morphogenesis1, mechano-transduction2, focal adhesion3, motility4,5, metastasis6 and drug delivery7,8,9,10. Techniques based on quasi-static atomic force microscopy techniques11,12,13,14,15,16,17 can map these properties, but they lack the spatial and temporal resolution that is needed to observe many of the relevant details. Here, we present a dynamic atomic force microscopy18,19,20,21,22,23,24,25,26,27,28 method to map quantitatively the nanomechanical properties of live cells with a throughput (measured in pixels/minute) that is ∼10–1,000 times higher than that achieved with quasi-static atomic force microscopy techniques. The local properties of a cell are derived from the 0th, 1st and 2nd harmonic components of the Fourier spectrum of the AFM cantilevers interacting with the cell surface. Local stiffness, stiffness gradient and the viscoelastic dissipation of live Escherichia coli bacteria, rat fibroblasts and human red blood cells were all mapped in buffer solutions. Our method is compatible with commercial atomic force microscopes and could be used to analyse mechanical changes in tumours, cells and biofilm formation with sub-10 nm detail.

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Figure 1: Harmonic content of the AFM microcantilever as it interacts with soft living cells.
Figure 2: Multi-harmonic images of E. coli bacteria.
Figure 3: Multi-harmonic images of rat fibroblasts.
Figure 4: Multi-harmonic images of a human red blood cell.
Figure 5: Tracking time-varying changes in the mechanical properties of rat fibroblasts.

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Acknowledgements

A.R. acknowledges financial support from the National Science Foundation (grant no. CMMI 0927648; programme manager, E. Misawa) and the Keeley visiting fellowship (awarded by Wadham Collage, University of Oxford) to support his stay at the University of Oxford. S.C. and A.R. also acknowledge financial support from the Engineering and Physical Sciences Research Council (EPSRC, grant no. EPSRC-EP/H043659/1). S.C. also acknowledges support from the Research Councils UK and Oxford Martin School.

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Author notes

  1. A. Raman and S. Trigueros: These authors contributed equally to this work

Authors and Affiliations

  1. School of Mechanical Engineering, Purdue University, West Lafayette, Indiana, USA

    A. Raman, A. Cartagena, M. Susilo & E. Nauman

  2. Department of Physics and Institute of Nanoscience for Medicine, Oxford Martin School, University of Oxford, Oxfordshire, UK

    S. Trigueros, A. P. Z. Stevenson & S. Antoranz Contera

  3. Birck Nanotechnology Center, Purdue University, West Lafayette, Indiana, USA

    A. Raman & A. Cartagena

  4. Weldon School of Biomedical Engineering, West Lafayette, Indiana, USA

    E. Nauman

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Contributions

A.R. and S.T. are lead authors and contributed equally to this work. A.R. discovered the important experimental channels for material contrast. S.T., A.C., A.S, A.R., E.N. and M.S. developed experimental protocols for sample preparation. A.R., S.T. and S.C. conceived and designed the experiments. A.R. developed the theory and A.C. performed the numerical simulations and developed the code to implement the theory on the acquired AFM images. A.R., A.C., A.S. and S.T. performed the experiments. A.R. and S.T. co-wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to A. Raman.

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The authors declare no competing financial interests.

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Raman, A., Trigueros, S., Cartagena, A. et al. Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. Nature Nanotech 6, 809–814 (2011). https://doi.org/10.1038/nnano.2011.186

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