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Localization-Based Super-Resolution Imaging of Cellular Structures

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Adhesion Protein Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1046))

Abstract

Fluorescence microscopy allows direct visualization of fluorescently tagged proteins within cells. However, the spatial resolution of conventional fluorescence microscopes is limited by diffraction to ~250 nm, prompting the development of super-resolution microscopy which offers resolution approaching the scale of single proteins, i.e., ~20 nm. Here, we describe protocols for single molecule localization-based super-resolution imaging, using focal adhesion proteins as an example and employing either photoswitchable fluorophores or photoactivatable fluorescent proteins. These protocols should also be easily adaptable to imaging a broad array of macromolecular assemblies in cells whose components can be fluorescently tagged and assemble into high density structures.

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Acknowledgments

PK is supported by the Singapore National Research Foundation under the NRF Fellowship (NRFF-2011-04). CMW is supported by the Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health. We thank Harald Hess and Gleb Shtengel (Howard Hughes Medical Institute, Janelia Farm Research Campus), and Michael Davidson (The Florida State University) for advice, equipment, reagents, and collaboration related to this work.

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Authors and Affiliations

  1. Department of Bioengineering, Mechanobiology Institute, National University of Singapore, Singapore, Singapore

    Pakorn Kanchanawong

  2. Cell Biology and Physiology Center, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Clare M. Waterman

Authors
  1. Pakorn Kanchanawong
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  2. Clare M. Waterman
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Editor information

Editors and Affiliations

  1. , Department of Oncology, University of Oxford, Roosevelt Drive, Oxford, OX3 7DQ, United Kingdom

    Amanda S. Coutts

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Kanchanawong, P., Waterman, C.M. (2013). Localization-Based Super-Resolution Imaging of Cellular Structures. In: Coutts, A. (eds) Adhesion Protein Protocols. Methods in Molecular Biology, vol 1046. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-538-5_4

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  • DOI: https://doi.org/10.1007/978-1-62703-538-5_4

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-537-8

  • Online ISBN: 978-1-62703-538-5

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