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Mapping molecular assemblies with fluorescence microscopy and object-based spatial statistics

Author

Listed:
  • Thibault Lagache

    (BioImage Analysis Unit. CNRS UMR 3691. 25 rue du Docteur Roux
    Columbia University)

  • Alexandre Grassart

    (Molecular Microbial Pathogenesis Unit. INSERM U1202. 28 rue du Docteur Roux)

  • Stéphane Dallongeville

    (BioImage Analysis Unit. CNRS UMR 3691. 25 rue du Docteur Roux)

  • Orestis Faklaris

    (Université Paris Diderot)

  • Nathalie Sauvonnet

    (Molecular Microbial Pathogenesis Unit. INSERM U1202. 28 rue du Docteur Roux)

  • Alexandre Dufour

    (BioImage Analysis Unit. CNRS UMR 3691. 25 rue du Docteur Roux)

  • Lydia Danglot

    (Team Membrane traffic in healthy and diseased brain)

  • Jean-Christophe Olivo-Marin

    (BioImage Analysis Unit. CNRS UMR 3691. 25 rue du Docteur Roux)

Abstract

Elucidating protein functions and molecular organisation requires to localise precisely single or aggregated molecules and analyse their spatial distributions. We develop a statistical method SODA (Statistical Object Distance Analysis) that uses either micro- or nanoscopy to significantly improve on standard co-localisation techniques. Our method considers cellular geometry and densities of molecules to provide statistical maps of isolated and associated (coupled) molecules. We use SODA with three-colour structured-illumination microscopy (SIM) images of hippocampal neurons, and statistically characterise spatial organisation of thousands of synapses. We show that presynaptic synapsin is arranged in asymmetric triangle with the 2 postsynaptic markers homer and PSD95, indicating a deeper localisation of homer. We then determine stoichiometry and distance between localisations of two synaptic vesicle proteins with 3D-STORM. These findings give insights into the protein organisation at the synapse, and prove the efficiency of SODA to quantitatively assess the geometry of molecular assemblies.

Suggested Citation

  • Thibault Lagache & Alexandre Grassart & Stéphane Dallongeville & Orestis Faklaris & Nathalie Sauvonnet & Alexandre Dufour & Lydia Danglot & Jean-Christophe Olivo-Marin, 2018. "Mapping molecular assemblies with fluorescence microscopy and object-based spatial statistics," Nature Communications, Nature, vol. 9(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:9:y:2018:i:1:d:10.1038_s41467-018-03053-x
    DOI: 10.1038/s41467-018-03053-x
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    Cited by:

    1. Simone Cauzzo & Ester Bruno & David Boulet & Paul Nazac & Miriam Basile & Alejandro Luis Callara & Federico Tozzi & Arti Ahluwalia & Chiara Magliaro & Lydia Danglot & Nicola Vanello, 2024. "A modular framework for multi-scale tissue imaging and neuronal segmentation," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    2. Sandra Mayr & Fabian Hauser & Sujitha Puthukodan & Markus Axmann & Janett Göhring & Jaroslaw Jacak, 2020. "Statistical analysis of 3D localisation microscopy images for quantification of membrane protein distributions in a platelet clot model," PLOS Computational Biology, Public Library of Science, vol. 16(6), pages 1-34, June.

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