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© Research
Publication : Nature communications

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

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Nature communications - 15 Feb 2018

Lagache T, Grassart A, Dallongeville S, Faklaris O, Sauvonnet N, Dufour A, Danglot L, Olivo-Marin JC

Link to Pubmed [PMID] – 29449608

Nat Commun 2018 Feb;9(1):698

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.

https://www.ncbi.nlm.nih.gov/pubmed/29449608