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Stochastic optical reconstruction microscopy-based relative localization analysis (STORM-RLA) for quantitative nanoscale assessment of spatial protein organization

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dc.contributor.authorVeeraraghavan, Rengasayeeen
dc.contributor.authorGourdie, Robert G.en
dc.contributor.departmentFralin Biomedical Research Instituteen
dc.contributor.departmentSchool of Biomedical Engineering and Sciencesen
dc.date.accessioned2017-06-07T20:45:14Zen
dc.date.available2017-06-07T20:45:14Zen
dc.date.issued2016-11-07en
dc.description.abstractThe spatial association between proteins is crucial to understanding how they function in biological systems. Colocalization analysis of fluorescence microscopy images is widely used to assess this. However, colocalization analysis performed on two-dimensional images with diffraction-limited resolution merely indicates that the proteins are within 200–300 nm of each other in the xy-plane and within 500–700 nm of each other along the z-axis. Here we demonstrate a novel three-dimensional quantitative analysis applicable to single-molecule positional data: stochastic optical reconstruction microscopy–based relative localization analysis (STORM-RLA). This method offers significant advantages: 1) STORM imaging affords 20-nm resolution in the xy-plane and <50 nm along the z-axis; 2) STORM-RLA provides a quantitative assessment of the frequency and degree of overlap between clusters of colabeled proteins; and 3) STORM-RLA also calculates the precise distances between both overlapping and nonoverlapping clusters in three dimensions. Thus STORM-RLA represents a significant advance in the high-throughput quantitative assessment of the spatial organization of proteins.en
dc.description.versionPublished versionen
dc.format.extent3583 - 3590 (8) page(s)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.1091/mbc.E16-02-0125en
dc.identifier.issn1059-1524en
dc.identifier.issue22en
dc.identifier.orcidGourdie, RG [0000-0001-6021-0796]en
dc.identifier.urihttp://hdl.handle.net/10919/77942en
dc.identifier.volume27en
dc.language.isoenen
dc.publisherAmerican Society for Cell Biologyen
dc.relation.urihttp://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000387391400019&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=930d57c9ac61a043676db62af60056c1en
dc.rightsCreative Commons Attribution-NonCommercial-ShareAlike 3.0 Unporteden
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/3.0/en
dc.subjectCell Biologyen
dc.subjectINTERCALATED DISCen
dc.subjectFLUORESCENCE MICROSCOPYen
dc.subjectCONDUCTIONen
dc.subjectTRACKINGen
dc.titleStochastic optical reconstruction microscopy-based relative localization analysis (STORM-RLA) for quantitative nanoscale assessment of spatial protein organizationen
dc.title.serialMolecular Biology of the Cellen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
pubs.organisational-group/Virginia Techen
pubs.organisational-group/Virginia Tech/All T&R Facultyen
pubs.organisational-group/Virginia Tech/Faculty of Health Sciencesen
pubs.organisational-group/Virginia Tech/University Research Institutesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciencesen
pubs.organisational-group/Virginia Tech/University Research Institutes/Fralin Life Sciences/Fralin Affiliated Facultyen
pubs.organisational-group/Virginia Tech/University Research Institutes/Virginia Tech Carilion Research Instituteen

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