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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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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