Holographic Photolysis for Multiple Cell Stimulation in Mouse Hippocampal Slices 英文参考文献.docVIP

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Holographic Photolysis for Multiple Cell Stimulation in Mouse Hippocampal Slices 英文参考文献.doc

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Holographic Photolysis for Multiple Cell Stimulation in Mouse Hippocampal Slices 英文参考文献

HolographicPhotolysisforMultipleCellStimulationin MouseHippocampalSlices MoradZahid1.,MateoVe′lez-Fort2.,EiriniPapagiakoumou1,CathieVentalon1,Mar?′aCeciliaAngulo2, ValentinaEmiliani1* 1Wavefront-EngineeringMicroscopyGroup,NeurophysiologyandNewMicroscopiesLaboratory,CNRSUMR8154,INSERMU603,UniversityParisDescartes,Paris,France, 2Neuron-GliaInteractionsGroup,NeurophysiologyandNewMicroscopiesLaboratory,CNRSUMR8154,INSERMU603,UniversityParisDescartes,Paris,France Abstract Background:Advancedlightmicroscopyofferssensitiveandnon-invasivemeanstoimageneuralactivityandtocontrol signalingwithphotolysablemoleculesand,recently,light-gatedchannels.Theseapproachesrequirepreciseandyetflexible light excitation patterns. For synchronous stimulation of subsets of cells, they also require large excitation areas with millisecondandmicrometricresolution.Wehaverecentlydevelopedanewmethodforsuchopticalcontrolusingaphase holographicmodulationofopticalwave-fronts,whichminimizespowerloss,enablesrapidswitchingbetweenexcitation patterns, and allows a true 3D sculpting of the excitation volumes. In previous studies we have used holographic photololysistocontrolglutamateuncagingonsingleneuronalcells.Here,weextendtheuseofholographicphotolysisfor theexcitationofmultipleneuronsandofglialcells. Methods/Principal Findings: The system combines a liquid crystal device for holographic patterned photostimulation, high-resolutionopticalimaging,theHiLomicroscopy,todefinethestimulatedregionsandaconventionalCa2+ imaging systemtodetectneuralactivity.Bymeansofelectrophysiologicalrecordingsandcalciumimaginginacutehippocampal slices,weshowthattheuseofexcitationpatternspreciselytailoredtotheshapeofmultipleneuronalsomatarepresentsa very efficient way for the simultaneous excitation of a group of neurons. In addition, we demonstrate that fast shaped illuminationpatternsalsoinducereliableresponsesinsingleglialcells. Conclusions/Significance: Weshowthatthemainadvantageofholographicilluminationisthatitallowsforaneffi