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dc.contributor.authorPeddie, Christopher J.
dc.contributor.authorDomart, Marie-Charlotte
dc.contributor.authorSnetkov, Xenia
dc.contributor.authorO'Toole, Peter
dc.contributor.authorLarijani, Banafshé ORCID
dc.contributor.authorWay, Michael
dc.contributor.authorCox, Susan
dc.contributor.authorCollinson, Lucy M.
dc.date.accessioned2019-01-16T11:06:32Z
dc.date.available2019-01-16T11:06:32Z
dc.date.issued2017-08
dc.identifier.citationJournal of Structural Biology 199(2) : 120-131 (2017)es_ES
dc.identifier.issn1047-8477
dc.identifier.issn1095-8657
dc.identifier.urihttp://hdl.handle.net/10810/30902
dc.description.abstractSuper-resolution light microscopy, correlative light and electron microscopy, and volume electron microscopy are revolutionising the way in which biological samples are examined and understood. Here, we combine these approaches to deliver super-accurate correlation of fluorescent proteins to cellular structures. We show that YFP and GFP have enhanced blinking properties when embedded in acrylic resin and imaged under partial vacuum, enabling in vacuo single molecule localisation microscopy. In conventional section-based correlative microscopy experiments, the specimen must be moved between imaging systems and/or further manipulated for optimal viewing. These steps can introduce undesirable alterations in the specimen, and complicate correlation between imaging modalities. We avoided these issues by using a scanning electron microscope with integrated optical microscope to acquire both localisation and electron microscopy images, which could then be precisely correlated. Collecting data from ultrathin sections also improved the axial resolution and signal-to-noise ratio of the raw localisation microscopy data. Expanding data collection across an array of sections will allow 3-dimensional correlation over unprecedented volumes. The performance of this technique is demonstrated on vaccinia virus (with YFP) and diacylglycerol in cellular membranes (with GFP).es_ES
dc.description.sponsorshipThis work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001999), the UK Medical Research Council (FC001999), and the Wellcome Trust (FC001999); and by the MRC, BBSRC and EPSRC under grant award MR/K01580X/1 to LMC and POT, and awards MR/K015664/1 and BB/K01563X/1 to SC. SC acknowledges support from a Royal Society University Research Fellowship. We would like to thank Sander den Hoedt (DELMIC B.V.) for useful discussions, and Bernd Rieger (T.U. Delft) for advice on using the FRC code.es_ES
dc.language.isoenges_ES
dc.publisherElsevieres_ES
dc.rightsinfo:eu-repo/semantics/openAccesses_ES
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/es/*
dc.subjectCLEMes_ES
dc.subjectILSEMes_ES
dc.subjectcorrelativees_ES
dc.subjectvolumees_ES
dc.subjectelectron microscopyes_ES
dc.subjectsuper-resolutiones_ES
dc.subjectintegratedes_ES
dc.subjectYFPes_ES
dc.subjectGFPes_ES
dc.subjectfluorescencees_ES
dc.subjectprotein localisationes_ES
dc.subject3-dimensionales_ES
dc.subjectblinkinges_ES
dc.subjectIn-resin fluorescencees_ES
dc.subjectvaccinia viruses_ES
dc.subjectlocalization microscopyes_ES
dc.subjectcryoelectron tomographyes_ES
dc.subjectstructural basises_ES
dc.subjectdynamicses_ES
dc.subjectphotoactivationes_ES
dc.subjectcellses_ES
dc.subjectlightes_ES
dc.subjectresolutiones_ES
dc.subjectnanoscopyes_ES
dc.titleCorrelative super-resolution fluorescence and electron microscopy using conventional fluorescent proteins in vacuoes_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.rights.holderThis is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)es_ES
dc.rights.holderAtribución 3.0 España*
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S104784771730093X?via%3Dihubes_ES
dc.identifier.doi10.1016/j.jsb.2017.05.013
dc.departamentoesMedicinaes_ES
dc.departamentoeuMedikuntzaes_ES


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This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
Except where otherwise noted, this item's license is described as This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)