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

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.
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Submitted on : Friday, June 28, 2019 - 1:52:37 PM
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Thibault Lagache, Alexandre Grassart, Stéphane Dallongeville, Orestis Faklaris, Nathalie Sauvonnet, et al.. Mapping molecular assemblies with fluorescence microscopy and object-based spatial statistics. Nature Communications, Nature Publishing Group, 2018, 9 (1), pp.698. ⟨10.1038/s41467-018-03053-x⟩. ⟨pasteur-02168087⟩

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