Targeted volume correlative light and electron microscopy of an environmental marine microorganism

Author:

Mocaer Karel12,Mizzon Giulia345,Gunkel Manuel6,Halavatyi Aliaksandr6ORCID,Steyer Anna7ORCID,Oorschot Viola3ORCID,Schorb Martin3ORCID,Le Kieffre Charlotte8,Yee Daniel P.8,Chevalier Fabien8,Gallet Benoit9ORCID,Decelle Johan8,Schwab Yannick13ORCID,Ronchi Paolo3ORCID

Affiliation:

1. European Molecular Biology Laboratory 1 Cell Biology and Biophysics Unit , , 69117 Heidelberg , Germany

2. Collaboration for joint PhD degree between the European Molecular Biology Laboratory and the Heidelberg University 2 , Faculty of Biosciences, 69120 Heidelberg , Germany

3. European Molecular Biology Laboratory 3 Electron Microscopy Core Facility , , 69117 Heidelberg , Germany

4. 4 Department of Infectious Diseases, Molecular Virology, CIID, 69120 Heidelberg, Germany

5. 5 German Center for Infection Research, Heidelberg partner site, 69120 Heidelberg, Germany

6. European Molecular Biology Laboratory 6 Advanced Light Microscopy Facility , , 69117 Heidelberg , Germany

7. EMBL Imaging Centre, European Molecular Biology Laboratory 7 , 69117 Heidelberg , Germany

8. Université Grenoble Alpes, CNRS, CEA, INRAe, IRIG-LPCV 8 , 38054 Grenoble , France

9. Institut de Biologie Structurale (IBS), Université Grenoble Alpes, CEA, CNRS 9 , 38000 Grenoble , France

Abstract

ABSTRACT Photosynthetic microalgae are responsible for an important fraction of CO2 fixation and O2 production on Earth. Three-dimensional (3D) ultrastructural characterization of these organisms in their natural environment can contribute to a deeper understanding of their cell biology. However, the low throughput of volume electron microscopy (vEM) methods along with the complexity and heterogeneity of environmental samples pose great technical challenges. In the present study, we used a workflow based on a specific electron microscopy sample preparation method compatible with both light and vEM imaging in order to target one cell among a complex natural community. This method revealed the 3D subcellular landscape of a photosynthetic dinoflagellate, which we identified as Ensiculifera tyrrhenica, with quantitative characterization of multiple organelles. We show that this cell contains a single convoluted chloroplast and show the arrangement of the flagellar apparatus with its associated photosensitive elements. Moreover, we observed partial chromatin unfolding, potentially associated with transcription activity in these organisms, in which chromosomes are permanently condensed. Together with providing insights in dinoflagellate biology, this proof-of-principle study illustrates an efficient tool for the targeted ultrastructural analysis of environmental microorganisms in heterogeneous mixes.

Funder

Centre National de la Recherche Scientifique

Institut National de la Santé et de la Recherche Médicale

Université Grenoble Alpes

European Molecular Biology Laboratory

European Marine Biological Resource Centre

Agence Nationale de la Recherche

Boehringer Ingelheim Stiftung

Publisher

The Company of Biologists

Subject

Cell Biology

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