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Using Correlative Light and Electron Microscopy to Study Zebrafish Vascular Morphogenesis

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Tissue Morphogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1189))

Abstract

Live imaging is extremely useful to characterize the dynamics of cellular events in vivo, yet it is limited in terms of spatial resolution. Correlative light and electron microscopy (CLEM) allows combining live confocal microscopy with electron microscopy (EM) for the characterization of biological samples at high temporal and spatial resolution. Here we describe a protocol allowing extracting endothelial cell ultrastructure after having imaged the same cell in its in vivo context through live confocal imaging during zebrafish embryonic development.

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Acknowledgements

We thank Irina Pokrovskaia for the idea to use malachite green. We thank Patrick Schultz and Corinne Crucifix for sharing equipment and help with electron tomography at IGBMC. We thank the IGBMC imaging center, in particular Coralie Spiegelhalter, Nadia Messaddeq, Pascal Kessler, Marc Koch and Didier Hentsch. We also thank Devrim Acehan, from the Electron Microscopy Core Facility at EMBL, for his significant help with the F30. This work was supported by HFSP, INSERM, la ligue contre le cancer, FRM, and the seventh framework program (MC-IRG256549 (JV) and MC-IEF254951 (JGG)).

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Authors and Affiliations

  1. Institute of Genetics and Molecular and Cellular Biology (IGBMC), CNRS/INSERM/UdS, 1 rue Laurent Fries, 67404, Illkirch, France

    Fabien Monduc & Julien Vermot Ph.D.

  2. Electron Microscopy Core Facility - Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117, Heidelberg, Germany

    Yannick Schwab

  3. The Microenvironmental Niche in Tumorigenesis and Targeted Therapy, Inserm U1109, LabEx Medalis, Fédération de Médecine Translationnelle de Strasbourg (FMTS), 67000, Strasbourg, France

    Jacky G. Goetz

Authors
  1. Jacky G. Goetz
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  2. Fabien Monduc
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  3. Yannick Schwab
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  4. Julien Vermot Ph.D.
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Corresponding author

Correspondence to Julien Vermot Ph.D. .

Editor information

Editors and Affiliations

  1. Chemical & Biological Engineering, Princeton University, New Jersey, USA

    Celeste M. Nelson

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Goetz, J.G., Monduc, F., Schwab, Y., Vermot, J. (2015). Using Correlative Light and Electron Microscopy to Study Zebrafish Vascular Morphogenesis. In: Nelson, C. (eds) Tissue Morphogenesis. Methods in Molecular Biology, vol 1189. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1164-6_3

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  • DOI: https://doi.org/10.1007/978-1-4939-1164-6_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1163-9

  • Online ISBN: 978-1-4939-1164-6

  • eBook Packages: Springer Protocols

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