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Plastids pp 113–122Cite as

Imaging Plastids in 2D and 3D: Confocal and Electron Microscopy

Part of the Methods in Molecular Biology book series (MIMB,volume 1829)

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.

Key words

  • Confocal imaging
  • Transmission electron microscopy (TEM)
  • Scanning electron microscopy (SEM)
  • Focused ion beam (FIB)
  • Tomography
  • Arabidopsis
  • Phaeodactylum tricornutum

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  • DOI: 10.1007/978-1-4939-8654-5_7
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References

  1. Gunning B, Koenig F, Govindjee G (2006) A dedication to pionners of research on chloroplast structure. In: Wise RR, Hoober JK (eds) The structure and function of plastids. Advances in photosynthesis and respiration, vol 23. Springer, Dordrecht, pp xxiii–xxxxi

    Google Scholar 

  2. Gris JBA (1857) Recherches microscopiques sur la chlorophylle. Ann Sci Nat Bot Ser IV 7:179–219

    Google Scholar 

  3. Trécul AAL (1858) Des formations vésiculaires dans les cellules végétales. Ann Sci Nat Bot Ser IV 8(20–163):205–382

    Google Scholar 

  4. Doutreligne J (1935) Note sur la structure des chloroplastes. Porc Akad Wetensch Amsterd 38:886–896

    Google Scholar 

  5. Kausche GA, Ruska H (1940) Zur Frage der Chloroplastenstruktur. Naturwiss 28:303–304

    CAS  Google Scholar 

  6. Menke W (1940) Untersuchungen über den Feinbau des Protoplasmas mit dem Universal-Elektronenmikroskop. Protoplasma 35:115–130

    CrossRef  CAS  Google Scholar 

  7. Steahelin LA (2003) Chloroplast structure: from chlorophyll granules to supra-molecular architecture of thylakoid membranes. Photosynth Res 76:185–196

    CrossRef  Google Scholar 

  8. Giannuzzi L, Stevie F (eds) (2005) Introduction to focus ion beam. Springer, New York

    Google Scholar 

  9. Kizilyaprak C, Loussert C, Daraspe J, Humbel BM (2013) Focused ion beam scanning electron microscopy in biology. Microsc Microanal 19:874–875

    CrossRef  Google Scholar 

  10. Narayan K, Subramaniam S (2015) Focused ion beams in biology. Nat Methods 12:1021–1031

    CrossRef  CAS  PubMed  Google Scholar 

  11. Berges J, Franklin PJ, Harrison PJ (2001) Evolution of an artificial seawater medium: improvements in enriched seawater, artificial water over the last two decades. J Phycol 37:1138–1145

    CrossRef  Google Scholar 

  12. Flori S, Jouneau PH, Finazzi G, Maréchal E, Falconet D (2016) Ultrastructure of the periplastidial compartment of the diatom Phaeodactylum tricornutum. Protist 167(3):254–267

    CrossRef  Google Scholar 

  13. Flori S, Jouneau PH, Bailleul B, Gallet B, Estrozi LF, Moriscot C et al (2017) Plastid thylakoid architecture optimizes photosynthesis in diatoms. Nat Commun 8:15885

    CrossRef  CAS  PubMed  PubMed Central  Google Scholar 

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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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Author notes

  1. Serena Flori

    Present address: Marine Biological Association of the UK Citadel Hill, Plymouth, UK

Authors and Affiliations

  1. Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, UMR5168, Université Grenoble Alpes, Grenoble, France

    Serena Flori, Giovanni Finazzi & Denis Falconet

  2. Laboratoire d’Etudes des Matériaux par Microscopie Avancée, Institut Nanosciences et Cryogénie-Service de Physique des Matériaux et Microstructures, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, INAC-MEM, Université Grenoble Alpes, Grenoble, France

    Pierre-Henri Jouneau

  3. Institut de Biologie Structurale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, IBS, Université Grenoble Alpes, Grenoble, France

    Benoit Gallet, Leandro F. Estrozi, Christine Moriscot & Guy Schoehn

Authors
  1. Serena Flori
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  2. Pierre-Henri Jouneau
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  3. Benoit Gallet
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  4. Leandro F. Estrozi
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  5. Christine Moriscot
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  6. Guy Schoehn
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  7. Giovanni Finazzi
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  8. Denis Falconet
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Corresponding author

Correspondence to Denis Falconet .

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Editors and Affiliations

  1. Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, Institut National Recherche Agronomique, UMR5168, Université Grenoble Alpes, Grenoble, France

    Eric Maréchal

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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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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8653-8

  • Online ISBN: 978-1-4939-8654-5

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