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Nanoscale live-cell imaging using hopping probe ion conductance microscopy

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A Corrigendum to this article was published on 03 September 2009

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Abstract

We describe hopping mode scanning ion conductance microscopy that allows noncontact imaging of the complex three-dimensional surfaces of live cells with resolution better than 20 nm. We tested the effectiveness of this technique by imaging networks of cultured rat hippocampal neurons and mechanosensory stereocilia of mouse cochlear hair cells. The technique allowed examination of nanoscale phenomena on the surface of live cells under physiological conditions.

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Figure 1: Principles of adaptive HPICM.
Figure 2: Visualization of vertically protruding mechanosensitive stereocilia of auditory hair cells.
Figure 3: HPICM images of live hippocampal neurons.

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Change history

  • 03 September 2009

    NOTE: In the version of this paper originally published, references to previous work on pulse mode SICM should have been included (Mann et al. J. Neurosci. Methods 116, 113–117, (2002) and Happel et al. J. Microsc. 212, 144–151 (2003)). These references were removed during shortening of the paper for publication and have been added back to the PDF and HTML versions of this article. The pulse mode technique reported in these previous papers has conceptual similarity to our hopping mode SICM, in that distance feedback control is not continuous; thus, it also solves the problem of probe-sample collision for large cellular structures. However, the pulse mode technique is considerably slower owing to a different feedback mechanism and does not perform at nanoscale resolution.

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Acknowledgements

This study was supported by the UK Biotechnology and Biological Sciences Research Council and Medical Research Council to Y.E.K. and (BB/D01817X/1) to G.W.J.M. and T.G.S., and by the US National Organization for Hearing Research Foundation, the Kentucky Science and Engineering Foundation (KSEF-148-502-07-215) and National Institute on Deafness and Other Communication Disorders, National Institutes of Health (DC008861 to G.I.F.).

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Author notes

  1. Pavel Novak and Chao Li: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Medicine, Imperial College London, London, UK

    Pavel Novak, Andrew I Shevchuk, Max J Lab & Yuri E Korchev

  2. Department of Cardiac Medicine, National Heart and Lung Institute, Imperial College London, London, UK

    Pavel Novak & Julia Gorelik

  3. Department of Chemistry, Cambridge University, Cambridge, UK

    Chao Li, Victor P Ostanin & David Klenerman

  4. Department of Physiology, University of Kentucky, Lexington, Kentucky, USA

    Ruben Stepanyan & Gregory I Frolenkov

  5. Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK

    Matthew Caldwell, Simon Hughes, Trevor G Smart & Guy W J Moss

  6. Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, UK

    Matthew Caldwell & Guy W J Moss

Authors
  1. Pavel Novak
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  2. Chao Li
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  3. Andrew I Shevchuk
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  4. Ruben Stepanyan
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  5. Matthew Caldwell
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  6. Simon Hughes
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  7. Trevor G Smart
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  8. Julia Gorelik
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  9. Victor P Ostanin
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  10. Max J Lab
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  11. Guy W J Moss
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  12. Gregory I Frolenkov
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  13. David Klenerman
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  14. Yuri E Korchev
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Contributions

P.N., C.L., A.I.S. and V.P.O. implemented experimental design into the hardware and the software; P.N., A.I.S., R.S., M.C. and S.H. performed the experiments; G.I.F., D.K., G.W.J.M., T.G.S., M.J.L., J.G. and Y.E.K. designed the study; P.N., G.I.F., D.K., G.W.J.M., T.G.S., M.J.L. and Y.E.K. wrote the paper.

Corresponding authors

Correspondence to Gregory I Frolenkov, David Klenerman or Yuri E Korchev.

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Competing interests

P.N. has a consultancy agreement with Ionscope Ltd., a small start-up company selling scanning ion conductance microscopes. D.K., M.J.L., A.I.S. and Y.E.K. are shareholders of Ionscope Ltd.

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Supplementary Figures 1–3, Supplementary Methods (PDF 7112 kb)

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Novak, P., Li, C., Shevchuk, A. et al. Nanoscale live-cell imaging using hopping probe ion conductance microscopy. Nat Methods 6, 279–281 (2009). https://doi.org/10.1038/nmeth.1306

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