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The Use of Correlative Light-Electron Microscopy (CLEM) to Study PINK1/Parkin-Mediated Mitophagy

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Mitophagy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1759))

Abstract

In this chapter we describe the use of correlative light-electron microscopy (CLEM) to study, in cultured cells, the turnover of damaged mitochondria by PINK1/Parkin-dependent mitophagy. CLEM combines the advantages of light microscopy, which allows to image and rapidly screen a large number of the cells, while electron microscopy provides high-resolution imaging of these selected cells and a detailed structural analysis of their cellular organelles. We describe in detail how to prepare the cell cultures for optimum preservation of their cellular ultrastructure for CLEM using the most suitable buffers, fixatives, and embedding resins. These protocols are applicable for detailed ultrastructural analysis in a wide variety of organisms and cells, ranging from prokaryotic bacteria to mammalian cells.

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Acknowledgements

We thank Dr. John Kendrick-Jones for his help in the preparation of this chapter, Dr. Antonina J. Kruppa for generating the HEK293 cells stably expressing HA-Parkin, and Dr. Nicholas A. Bright for critical reading of the manuscript. This work was supported by Medical Research Council (MR/K000888/1).

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Authors and Affiliations

  1. Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK

    Chieko Kishi-Itakura & Folma Buss

Authors
  1. Chieko Kishi-Itakura
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  2. Folma Buss
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Corresponding authors

Correspondence to Chieko Kishi-Itakura or Folma Buss .

Editor information

Editors and Affiliations

  1. Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan

    Nobutaka Hattori

  2. Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan

    Shinji Saiki

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Kishi-Itakura, C., Buss, F. (2017). The Use of Correlative Light-Electron Microscopy (CLEM) to Study PINK1/Parkin-Mediated Mitophagy. In: Hattori, N., Saiki, S. (eds) Mitophagy. Methods in Molecular Biology, vol 1759. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_8

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  • DOI: https://doi.org/10.1007/7651_2017_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7749-9

  • Online ISBN: 978-1-4939-7750-5

  • eBook Packages: Springer Protocols

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