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dc.contributor.authorDurisic, Nela
dc.contributor.authorLaparra-Cuervo, Lara
dc.contributor.authorSandoval-Álvarez, Ángel
dc.contributor.authorBorbely, Joseph Steven
dc.contributor.authorLakadamyali, Melike
dc.contributor.otherUniversitat Politècnica de Catalunya. Institut de Ciències Fotòniques
dc.date.accessioned2016-07-19T08:07:05Z
dc.date.available2016-07-19T08:07:05Z
dc.date.issued2014-01-05
dc.identifier.issn1548-7091
dc.identifier.urihttp://hdl.handle.net/2117/88877
dc.description.abstractPhotoswitchable fluorescent probes are central to localization-based super-resolution microscopy. Among these probes, fluorescent proteins are appealing because they are genetically encoded. Moreover, the ability to achieve a 1:1 labeling ratio between the fluorescent protein and the protein of interest makes these probes attractive for quantitative single-molecule counting. The percentage of fluorescent protein that is photoactivated into a fluorescently detectable form (i.e., the photoactivation efficiency) plays a crucial part in properly interpreting the quantitative information. It is important to characterize the photoactivation efficiency at the single-molecule level under the conditions used in super-resolution imaging. Here, we used the human glycine receptor expressed in Xenopus oocytes and stepwise photobleaching or single-molecule counting photoactivated localization microcopy (PALM) to determine the photoactivation efficiency of fluorescent proteins mEos2, mEos3.1, mEos3.2, Dendra2, mClavGR2, mMaple, PA-GFP and PA-mCherry. This analysis provides important information that must be considered when using these fluorescent proteins in quantitative super-resolution microscopy.
dc.format.extent7 p.
dc.language.isoeng
dc.publisherNature
dc.rightsAttribution-NonCommercial-NoDerivs 3.0 Spain
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
dc.subjectÀrees temàtiques de la UPC::Física
dc.subject.lcshfluorescence
dc.subject.otherfluorescent protein
dc.titleSingle molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate
dc.typeArticle
dc.subject.lemacFluorescència
dc.description.peerreviewedPeer Reviewed
dc.relation.publisherversionhttp://doi.org/10.1038/nmeth.2784
dc.rights.accessOpen Access
dc.description.versionPostprint (author's final draft)
upcommons.citation.pubplaceUSA
upcommons.citation.publicationNameNATURE METHODS
upcommons.citation.volume11
upcommons.citation.startingPage156
upcommons.citation.endingPage162


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