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

Thibault Lagache, Alexandre Grassart, Stéphane Dallongeville, Orestis Faklaris, Nathalie Sauvonnet, Alexandre Dufour, Lydia Danglot () and Jean-Christophe Olivo-Marin ()
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Thibault Lagache: BioImage Analysis Unit. CNRS UMR 3691. 25 rue du Docteur Roux
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

Nature Communications, 2018, vol. 9, issue 1, 1-15

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-03053-x

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