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Fluorescence Imaging of Osteoclasts Using Confocal Microscopy

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Bone Research Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 816))

Abstract

In order to understand osteoclast cell biology, it is necessary to culture these cells on a physiological ­substrate that they can resorb in vitro, such as bone or dentine. However, this creates problems for analysis by fluorescence microscopy, due to the depth of the sample under investigation. By virtue of its optical sectioning capabilities, confocal microscopy is ideal for analysis of such samples, enabling precise intracellular localisation of proteins in resorbing osteoclasts to be determined. Moreover, by taking a series of images in the axial dimension, it is possible to create axial section views and to reconstruct 3D images of the osteoclasts, enabling the spatial organisation of the structures of interest to be more easily discerned.

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Author information

Authors and Affiliations

  1. Musculoskeletal Research Programme, Division of Applied Medicine, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen, UK

    Fraser P. Coxon

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  1. Fraser P. Coxon
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Correspondence to Fraser P. Coxon .

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Editors and Affiliations

  1. Institute of Medical Sciences, University of Aberdeen, Forester Hill, Aberdeen, AB25 2ZD, United Kingdom

    Miep H. Helfrich

  2. Institute of Genetics and Molecular Medi, Rheumatic Diseases Unit, University of Edinburgh, Crewe Road South, Edinburgh, EH4 2XU, United Kingdom

    Stuart H. Ralston

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Coxon, F.P. (2012). Fluorescence Imaging of Osteoclasts Using Confocal Microscopy. In: Helfrich, M., Ralston, S. (eds) Bone Research Protocols. Methods in Molecular Biology, vol 816. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-415-5_25

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  • DOI: https://doi.org/10.1007/978-1-61779-415-5_25

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-414-8

  • Online ISBN: 978-1-61779-415-5

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