On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed Fluorescent Synaptic Markers 英文参考文献.docVIP

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On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed Fluorescent Synaptic Markers 英文参考文献.doc

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On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed Fluorescent Synaptic Markers 英文参考文献

OnOpticalDetectionofDenselyLabeledSynapsesin NeuropilandMappingConnectivitywith CombinatoriallyMultiplexedFluorescentSynaptic Markers YuriyMishchenko* DepartmentofStatisticsandCenterforTheoreticalNeuroscience,ColumbiaUniversity,NewYork,NewYork,UnitedStatesofAmerica Abstract Weproposeanewmethodformappingneuralconnectivityoptically,byutilizingCre/LoxsystemBrainbowtotagsynapses ofdifferentneuronswithrandommixturesofdifferentfluorophores,suchasGFP,YFP,etc.,andthendetectingpatternsof fluorophoresatdifferentsynapsesusinglightmicroscopy(LM).Suchpatternswillimmediatelyreportthepre-andpost- synapticcellsateachsynapticconnection,withouttracingneuralprojectionsfromindividualsynapsestocorrespondingcell bodies. We simulate fluorescence from a population of densely labeled synapses in a block of hippocampal neuropil, completelyreconstructedfromelectronmicroscopydata,andshowthathigh-endLMisabletodetectsuchpatternswith over95%accuracy.Weconclude,therefore,thatwiththedescribedapproachneuralconnectivityinmacroscopicallylarge neuralcircuitscanbemappedwithgreataccuracy,inscalablemanner,usingfastopticaltools,andstraightforwardimage processing.Relyingonanelectronmicroscopydataset,wealsoderiveandexplicitlyenumeratetheconditionsthatshould bemettoallowsynapticconnectivitystudieswithhigh-resolutionopticaltools. Citation: Mishchenko Y (2010) On Optical Detection of Densely Labeled Synapses in Neuropil and Mapping Connectivity with Combinatorially Multiplexed FluorescentSynapticMarkers.PLoSONE5(1):e8853.doi:10.1371/journal.pone.0008853 Editor:DavidFinkelstein,TheMentalHealthResearchInstituteofVictoria,Australia ReceivedOctober19,2009;AcceptedDecember24,2009;PublishedJanuary22,2010 Copyright:?2010YuriyMishchenko.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Theauthorhasnosupportorfundingtoreport.Thestudywasnotspecificallyfinanci