Lateral diffusion on tubular membranes: quantification of measurements bias.

Single Particle Tracking (SPT) is a powerful technique for the analysis of the lateral diffusion of the lipid and protein components of biological membranes. In neurons, SPT allows the study of the real-time dynamics of receptors for neurotransmitters that diffuse continuously in and out synapses. I...

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Main Authors: Marianne Renner, Yegor Domanov, Fanny Sandrin, Ignacio Izeddin, Patricia Bassereau, Antoine Triller
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2011-01-01
Series:PLoS ONE
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21980531/pdf/?tool=EBI
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author Marianne Renner
Yegor Domanov
Fanny Sandrin
Ignacio Izeddin
Patricia Bassereau
Antoine Triller
author_facet Marianne Renner
Yegor Domanov
Fanny Sandrin
Ignacio Izeddin
Patricia Bassereau
Antoine Triller
author_sort Marianne Renner
collection DOAJ
description Single Particle Tracking (SPT) is a powerful technique for the analysis of the lateral diffusion of the lipid and protein components of biological membranes. In neurons, SPT allows the study of the real-time dynamics of receptors for neurotransmitters that diffuse continuously in and out synapses. In the simplest case where the membrane is flat and is parallel to the focal plane of the microscope the analysis of diffusion from SPT data is relatively straightforward. However, in most biological samples the membranes are curved, which complicates analysis and may lead to erroneous conclusions as for the mode of lateral diffusion. Here we considered the case of lateral diffusion in tubular membranes, such as axons, dendrites or the neck of dendritic spines. Monte Carlo simulations allowed us to evaluate the error in diffusion coefficient (D) calculation if the curvature is not taken into account. The underestimation is determined by the diameter of the tubular surface, the frequency of image acquisition and the degree of mobility itself. We found that projected trajectories give estimates that are 25 to 50% lower than the real D in case of 2D-SPT over the tubular surface. The use of 3D-SPT improved the measurements if the frequency of image acquisition was fast enough in relation to the mobility of the molecules and the diameter of the tube. Nevertheless, the calculation of D from the components of displacements in the axis of the tubular structure gave accurate estimate of D, free of geometrical artefacts. We show the application of this approach to analyze the diffusion of a lipid on model tubular membranes and of a membrane-bound GFP on neurites from cultured rat hippocampal neurons.
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spelling doaj.art-308033a427ae445c84731e6319f9e2402022-12-21T23:10:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0169e2573110.1371/journal.pone.0025731Lateral diffusion on tubular membranes: quantification of measurements bias.Marianne RennerYegor DomanovFanny SandrinIgnacio IzeddinPatricia BassereauAntoine TrillerSingle Particle Tracking (SPT) is a powerful technique for the analysis of the lateral diffusion of the lipid and protein components of biological membranes. In neurons, SPT allows the study of the real-time dynamics of receptors for neurotransmitters that diffuse continuously in and out synapses. In the simplest case where the membrane is flat and is parallel to the focal plane of the microscope the analysis of diffusion from SPT data is relatively straightforward. However, in most biological samples the membranes are curved, which complicates analysis and may lead to erroneous conclusions as for the mode of lateral diffusion. Here we considered the case of lateral diffusion in tubular membranes, such as axons, dendrites or the neck of dendritic spines. Monte Carlo simulations allowed us to evaluate the error in diffusion coefficient (D) calculation if the curvature is not taken into account. The underestimation is determined by the diameter of the tubular surface, the frequency of image acquisition and the degree of mobility itself. We found that projected trajectories give estimates that are 25 to 50% lower than the real D in case of 2D-SPT over the tubular surface. The use of 3D-SPT improved the measurements if the frequency of image acquisition was fast enough in relation to the mobility of the molecules and the diameter of the tube. Nevertheless, the calculation of D from the components of displacements in the axis of the tubular structure gave accurate estimate of D, free of geometrical artefacts. We show the application of this approach to analyze the diffusion of a lipid on model tubular membranes and of a membrane-bound GFP on neurites from cultured rat hippocampal neurons.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21980531/pdf/?tool=EBI
spellingShingle Marianne Renner
Yegor Domanov
Fanny Sandrin
Ignacio Izeddin
Patricia Bassereau
Antoine Triller
Lateral diffusion on tubular membranes: quantification of measurements bias.
PLoS ONE
title Lateral diffusion on tubular membranes: quantification of measurements bias.
title_full Lateral diffusion on tubular membranes: quantification of measurements bias.
title_fullStr Lateral diffusion on tubular membranes: quantification of measurements bias.
title_full_unstemmed Lateral diffusion on tubular membranes: quantification of measurements bias.
title_short Lateral diffusion on tubular membranes: quantification of measurements bias.
title_sort lateral diffusion on tubular membranes quantification of measurements bias
url https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21980531/pdf/?tool=EBI
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