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Electron Microscopy pp 559–592Cite as

Correlative Light and Electron Microscopy Using Immunolabeled Sections

Part of the Methods in Molecular Biology book series (MIMB,volume 1117)

Abstract

In correlative microscopy, light microscopy provides the overview and orientation of the complex cells and tissue, while electron microscopy offers the detailed localization and correlation of subcellular structures. In this chapter we offer detailed high-quality electron microscopical preparation methods for optimum preservation of the cellular ultrastructure. From such preparations serial thin sections are collected and used for comparative histochemical, immunofluorescence, and immunogold staining.

In light microscopy histological stains identify the orientation of the sample and immunofluorescence labeling facilitates to find the region of interest, namely, the labeled cells expressing the macromolecule under investigation. Sections, labeled with immunogold are analyzed by electron microscopy in order to identify the label within the cellular architecture at high resolution.

Key words

  • Immunolabeling
  • Immunofluorescence
  • Immunogold
  • Light microscopy
  • Transmission electron microscopy
  • Scanning electron microscopy
  • Resin embedding
  • Methacrylate
  • Epoxy
  • Tokuyasu cryosectioning
  • Correlative microscopy
  • Array tomography

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Acknowledgments

The zebra fish specimen and the anti-prolactin serum were provided by Dr. Matthias Hammerschmidt (University of Cologne). Dr. Roger Wepf (EMEZ, ETH Zürich) recommended to use ITO-coated coverslips for SEM and provided such coverslips for pilot experiments. We thank Mrs. Brigitte Sailer for technical support, Mrs. Gertrud Scheer for excellent photographic work, and Dr. Céline Loussert and Dr. York-Dieter Stierhof for their valuable comments on the manuscript. We also would like to thank the financial support by the Faculty of Biology and Medicine of the University of Lausanne and by the R'Equip grant 316030_128692 of the Swiss National Science Foundation.

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Authors and Affiliations

  1. Max Planck Institute for Developmental Biology, Tübingen, Germany

    Heinz Schwarz

  2. Electron Microscopy Facility, Biophore, University of Lausanne, Lausanne, Switzerland

    Bruno M. Humbel

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  1. Heinz Schwarz
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  2. Bruno M. Humbel
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Editors and Affiliations

  1. University of Western Australia, Crawley, Australia

    John Kuo

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Schwarz, H., Humbel, B.M. (2014). Correlative Light and Electron Microscopy Using Immunolabeled Sections. In: Kuo, J. (eds) Electron Microscopy. Methods in Molecular Biology, vol 1117. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-776-1_25

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  • DOI: https://doi.org/10.1007/978-1-62703-776-1_25

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-775-4

  • Online ISBN: 978-1-62703-776-1

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