Probing Cellular Dynamics with a Chemical Signal Generator 英文参考文献.docVIP

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Probing Cellular Dynamics with a Chemical Signal Generator 英文参考文献.doc

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Probing Cellular Dynamics with a Chemical Signal Generator 英文参考文献

ProbingCellularDynamicswithaChemicalSignal Generator BrandonKuczenski1,WarrenC.Ruder2,WilliamC.Messner1*,PhilipR.LeDuc1,2* 1Department of Mechanical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, United States of America, 2Department of Biomedical Engineering, CarnegieMellonUniversity,Pittsburgh,Pennsylvania,UnitedStatesofAmerica Abstract Observationsofmaterialandcellularsystemsinresponsetotime-varyingchemicalstimulicanaidtheanalysisofdynamic processes. We describe a microfluidic ‘‘chemical signal generator,’’ a technique to apply continuously varying chemical concentrationwaveformstoarbitrarylocationsinamicrofluidicchannelthroughfeedbackcontroloftheinterfacebetween parallel laminar (co-flowing) streams. As the flow rates of the streams are adjusted, the channel walls are exposed to a chemical environment that shifts between the individual streams. This approach can be used to probe the dynamic behaviorofobjectsorsubstancesadherenttotheinteriorofthechannel.Todemonstratethetechnique,weexposedlive fibroblastcellstoionomycin,amembrane-permeablecalciumionophore,whileassayingcytosoliccalciumconcentration. Throughthemanipulationofthelaminarflowinterface,weexposedthecells’endogenouscalciumhandlingmachineryto spatially-containeddiscreteandoscillatoryintracellulardisturbances,whichwereobservedtoelicitaregulatoryresponse. The spatiotemporal precision of the generated signals opens avenues to previously unapproachable areas for potential investigationofcellsignalingandmaterialbehavior. Citation:KuczenskiB,RuderWC,MessnerWC,LeDucPR(2009)ProbingCellularDynamicswithaChemicalSignalGenerator.PLoSONE4(3):e4847.doi:10.1371/ journal.pone.0004847 Editor:MauricioRojas,EmoryUniversity,UnitedStatesofAmerica ReceivedSeptember17,2008;AcceptedFebruary4,2009;PublishedMarch16,2009 Copyright: ? 2009 Kuczenski et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproduct