Automated quantification of protein periodic nanostructures in fluorescence nanoscopy images: Abundance and regularity of neuronal spectrin membrane-associated skeleton

Fluorescence nanoscopy imaging permits the observation of periodic supramolecular protein structures in their natural environment, as well as the unveiling of previously unknown protein periodic structures. Deciphering the biological functions of such protein nanostructures requires systematic and q...

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Bibliographic Details
Authors: Barabas, Federico Martín, Masullo, Luciano Andrés, Bordenave, Martín Diego, Giusti, Sebastian Alejandro, Unsain, Nicolas, Refojo, Damian, Caceres, Alfredo Oscar, Stefani, Fernando Daniel
Format: article
Status:Published version
Publication Date:2017
Country:Argentina
Institution:Consejo Nacional de Investigaciones Científicas y Técnicas
Repository:CONICET Digital (CONICET)
Language:English
OAI Identifier:oai:ri.conicet.gov.ar:11336/54317
Online Access:http://hdl.handle.net/11336/54317
Access Level:Open access
Keyword:PROTEIN SELF-ASSEMBLY
SUPER-RESOLUTION
NEURON
CYTOSKELETON
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
Description
Summary:Fluorescence nanoscopy imaging permits the observation of periodic supramolecular protein structures in their natural environment, as well as the unveiling of previously unknown protein periodic structures. Deciphering the biological functions of such protein nanostructures requires systematic and quantitative analysis of large number of images under different experimental conditions and specific stimuli. Here we present a method and an open source software for the automated quantification of protein periodic structures in super-resolved images. Its performance is demonstrated by analyzing the abundance and regularity of the spectrin membrane-associated periodic skeleton (MPS) in hippocampal neurons of 2 to 40 days in vitro, imaged by STED and STORM nanoscopy. The automated analysis reveals that both the abundance and the regularity of the MPS increase over time and reach maximum plateau values after 14 DIV. A detailed analysis of the distributions of correlation coefficients provides indication of dynamical assembly and disassembly of the MPS.

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