A Microscope Automated Fluidic System to Study Bacterial Processes in Real Time 英文参考文献.docVIP

AMicroscopeAutomatedFluidicSystemtoStudy BacterialProcessesinRealTime AdrienDucret,EtienneMaisonneuve,PhilippeNotareschi,AlainGrossi,Ta?mMignot*,SamDukan* Aix Marseille Universite′ - Laboratoire de Chimie Bacte′rienne (UPR 9043) - Institut de Microbiologie de la Me′diterrane′e (IFR 88) - CNRS, 31, Chemin Joseph Aiguier, Marseille,France Abstract Most time lapse microscopy experiments studying bacterial processes ie growth, progression through the cell cycle and motility have been performed on thin nutrient agar pads. An important limitation of this approach is that dynamic perturbationsoftheexperimentalconditionscannotbeeasilyperformed.Ineukaryoticcellbiology,fluidicapproacheshave beenlargelyusedtostudytheimpactofrapidenvironmentalperturbationsonlivecellsandinrealtime.However,allthese approaches are not easily applicable to bacterial cells because the substrata are in all cases specific and also because microfluidicsnanotechnologyrequiresacomplexlithographyforthestudyofmicrometersizedbacterialcells.Infact,in manycasesagaristheexperimentalsolidsubstratumonwhichbacteriacanmoveorevengrow.Forthesereasons,we designedanovelhybridmicrofluidicdevicethatcombinesathinagarpadandacustomflowchamber.Bystudyingseveral examples, we show that this system allows real time analysis of a broad array of biological processes such as growth, developmentandmotility.Thus,theflowchambersystemwillbeanessentialtooltostudyanyprocessthattakeplaceon anagarsurfaceatthesinglecelllevel. Citation:DucretA,MaisonneuveE,NotareschiP,GrossiA,MignotT,etal.(2009)AMicroscopeAutomatedFluidicSystemtoStudyBacterialProcessesinReal Time.PLoSONE4(9):e7282.doi:10.1371/journal.pone.0007282 Editor:ChristopheHerman,BaylorCollegeofMedicine,UnitedStatesofAmerica ReceivedJuly31,2009;AcceptedSeptember8,2009;PublishedSeptember30,2009 Copyright: ? 2009 Ducret et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproduc