Barcoding T Cell Calcium Response Diversity with Methods for Automated and Accurate Analysis of Cell Signals (MAAACS).docVIP

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Barcoding T Cell Calcium Response Diversity with Methods for Automated and Accurate Analysis of Cell Signals (MAAACS).doc

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Barcoding T Cell Calcium Response Diversity with Methods for Automated and Accurate Analysis of Cell Signals (MAAACS)

BarcodingTCellCalciumResponseDiversitywith MethodsforAutomatedandAccurateAnalysisofCell Signals(MAAACS) AudreySalles1,2,3.¤a,CyrilleBillaudeau1,2,3.¤b,ArnauldSerge′1,2,3¤c*,Anne-MarieBernard1,2,3 Marie-ClairePhe′lipot1,2,3,NicolasBertaux4,5,MathieuFallet1,2,3,PierreGrenot1,2,3,DidierMarguet1,2,3 Hai-TaoHe1,2,3,YannickHamon1,2,3 , , * 1Centred’ImmunologiedeMarseille-Luminy(CIML),Aix-MarseilleUniversity,UM2,Marseille,France,2InstitutNationaldelaSante′ etdelaRechercheMe′dicale(Inserm), U1104,Marseille,France,3CentreNationaldelaRechercheScientifique(CNRS)UMR7280,Marseille,France,4InstitutFresnel,CentreNationaldelaRechercheScientifique (CNRS)UMR7249,Marseille,France,5E′coleCentraleMarseille,Technopo?ledeCha?teau-Gombert, Marseille,France Abstract WeintroduceaseriesofexperimentalproceduresenablingsensitivecalciummonitoringinTcellpopulationsbyconfocal video-microscopy. Tracking and post-acquisition analysis was performed using Methods for Automated and Accurate Analysis of Cell Signals (MAAACS), a fully customized program that associates a high throughput tracking algorithm, an intuitive reconnection routine and a statistical platform to provide, at a glance, the calcium barcode of a population of individualT-cells.Combinedwithasensitivecalciumprobe,thismethodallowedustounraveltheheterogeneityinshape andintensityofthecalciumresponseinTcellpopulationsandespeciallyinnaiveTcells,whichdisplayintracellularcalcium oscillationsuponstimulationbyantigenpresentingcells. Citation:SallesA,BillaudeauC,Serge′ A,BernardA-M,Phe′lipot M-C,etal.(2013)BarcodingTCellCalciumResponseDiversitywithMethodsforAutomatedand AccurateAnalysisofCellSignals(MAAACS).PLoSComputBiol9(9):e1003245.doi:10.1371/journal.pcbi.1003245 Editor:Gre′goireAltan-Bonnet,MemorialSloan-KetteringCancerCenter,UnitedStatesofAmerica ReceivedJanuary30,2013;AcceptedAugust15,2013;PublishedSeptember26,2013 Copyright: ? 2013 Salles et al. This is an open-access article distributed under the terms of the Creative Co