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dc.contributor.authorGhosh, Mitrajit
dc.contributor.authorvan den Akker, Nynke M. S.
dc.contributor.authorWijnands, Karolina A. P.
dc.contributor.authorPoeze, Martijn
dc.contributor.authorWeber, Christian
dc.contributor.authorMcQuade, Lindsey E.
dc.contributor.authorPluth, Michael D.
dc.contributor.authorLippard, Stephen J.
dc.contributor.authorPost, Mark J.
dc.contributor.authorMolin, Daniel G. M.
dc.contributor.authorvan Zandvoort, Marc A. M. J.
dc.date.accessioned2014-01-06T20:07:10Z
dc.date.available2014-01-06T20:07:10Z
dc.date.issued2013-09
dc.date.submitted2013-05
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/1721.1/83520
dc.description.abstractTo study the role and (sub) cellular nitric oxide (NO) constitution in various disease processes, its direct and specific detection in living cells and tissues is a major requirement. Several methods are available to measure the oxidation products of NO, but the detection of NO itself has proved challenging. We visualized NO production using a NO-sensitive copper-based fluorescent probe (Cu [subscript 2]FL2E) and two-photon laser scanning microscopy (TPLSM). Cu [subscript 2]FL2E demonstrated high sensitivity and specificity for NO synthesis, combined with low cytotoxicity. Furthermore, Cu [subscript 2]FL2E showed superior sensitivity over the conventionally used Griess assay. NO specificity of Cu [subscript 2]FL2E was confirmed in vitro in human coronary arterial endothelial cells and porcine aortic endothelial cells using various triggers for NO production. Using TPLSM on ex vivo mounted murine carotid artery and aorta, the applicability of the probe to image NO production in both endothelial cells and smooth muscle cells was shown. NO-production and time course was detected for multiple stimuli such as flow, acetylcholine and hydrogen peroxide and its correlation with vasodilation was demonstrated. NO-specific fluorescence and vasodilation was abrogated in the presence of NO-synthesis blocker L-NAME. Finally, the influence of carotid precontraction and vasorelaxation validated the functional properties of vessels. Specific visualization of NO production in vessels with Cu [subscript 2]FL2E-TPLSM provides a valid method for studying spatial-temporal synthesis of NO in vascular biology at an unprecedented level. This approach enables investigation of the pathways involved in the complex interplay between NO and vascular (dys) function.en_US
dc.description.sponsorshipNational Science Foundation (U.S.) (Grant CHE-0907905)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Grant K99GM092970)en_US
dc.language.isoen_US
dc.publisherPublic Library of Scienceen_US
dc.relation.isversionofhttp://dx.doi.org/10.1371/journal.pone.0075331en_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.5/en_US
dc.sourcePLoSen_US
dc.titleSpecific Visualization of Nitric Oxide in the Vasculature with Two-Photon Microscopy Using a Copper Based Fluorescent Probeen_US
dc.typeArticleen_US
dc.identifier.citationGhosh, Mitrajit, Nynke M. S. van den Akker, Karolina A. P. Wijnands, Martijn Poeze, Christian Weber, Lindsey E. McQuade, Michael D. Pluth, et al. “Specific Visualization of Nitric Oxide in the Vasculature with Two-Photon Microscopy Using a Copper Based Fluorescent Probe.” Edited by David D. Roberts. PLoS ONE 8, no. 9 (September 23, 2013): e75331.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemistryen_US
dc.contributor.mitauthorPluth, Michael D.en_US
dc.contributor.mitauthorLippard, Stephen J.en_US
dc.relation.journalPLoS ONEen_US
dc.eprint.versionFinal published versionen_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsGhosh, Mitrajit; van den Akker, Nynke M. S.; Wijnands, Karolina A. P.; Poeze, Martijn; Weber, Christian; McQuade, Lindsey E.; Pluth, Michael D.; Lippard, Stephen J.; Post, Mark J.; Molin, Daniel G. M.; van Zandvoort, Marc A. M. J.en_US
dc.identifier.orcidhttps://orcid.org/0000-0002-2693-4982
mit.licensePUBLISHER_CCen_US
mit.metadata.statusComplete


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