Abstract
Internal chloroplast structures present complex and various characteristics, which are still largely undetermined due to insufficient imaging investigation. Information on chloroplast morphology has traditionally been collected using light microscopy (LM), confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) techniques. However, recent technological progresses in the field of microscopy have made it possible to visualize the internal structure of chloroplast in far greater detail and in 3D. Here we recapitulate protocols to visualize chloroplasts from Arabidopsis leaves and Phaeodactylum tricornutum cells with confocal and transmission electron microscopy together with a new technique using a focused ion beam-scanning electron microscope (FIB-SEM) allowing for 3D imaging.
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Acknowledgments
Didier Grunwald and Jean-Philippe Kleman are acknowledged for their help with confocal microscopy. We are grateful to Maryse Block for critical reading of the manuscript. DF would like to thank Pr. Paul Mandaron for introducing him to plant imaging. Presented examples were obtained with support from the CEA DRF-Impulsion Fib-Bio program 2016 and the French National Research Agency (ANR-10-LABEX-04 GRAL Labex, Grenoble Alliance for Integrated Structural Cell Biology).
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Flori, S. et al. (2018). Imaging Plastids in 2D and 3D: Confocal and Electron Microscopy. In: Maréchal, E. (eds) Plastids. Methods in Molecular Biology, vol 1829. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8654-5_7
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DOI: https://doi.org/10.1007/978-1-4939-8654-5_7
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