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

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...

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Bibliographic Details
Published in:Nature communications 2018-02, Vol.9 (1), p.698-15, Article 698
Main Authors: Lagache, Thibault, Grassart, Alexandre, Dallongeville, Stéphane, Faklaris, Orestis, Sauvonnet, Nathalie, Dufour, Alexandre, Danglot, Lydia, Olivo-Marin, Jean-Christophe
Format: Article
Language:English
Subjects:
14/19
14/35
14/63
631/114/1564
631/114/2415
631/1647/245/2225
631/1647/328/2238
Assemblies
Cellular Biology
Computer Science
Fluorescence
Fluorescence microscopy
Hippocampus
Humanities and Social Sciences
Image Processing
Life Sciences
Localization
Microscopy
multidisciplinary
Peptide mapping
Postsynaptic density proteins
Proteins
Science
Science (multidisciplinary)
Spatial analysis
Spatial distribution
Statistical analysis
Statistical methods
Statistics
Stoichiometry
Synapses
Synapsin
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Summary: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. Elucidating molecular organisation requires precise localisation and analysis. Here the authors develop SODA software for automatic and quantitative mapping of statistically coupled molecules, and use it to unravel spatial organisation of thousands of synaptic proteins in SIM and 3DSTORM microscopy.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-018-03053-x