Single Cell Analysis of Yeast Replicative Aging Using a New Generation of Microfluidic Device 英文参考文献.docVIP

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Single Cell Analysis of Yeast Replicative Aging Using a New Generation of Microfluidic Device 英文参考文献.doc

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Single Cell Analysis of Yeast Replicative Aging Using a New Generation of Microfluidic Device 英文参考文献

SingleCellAnalysisofYeastReplicativeAgingUsinga NewGenerationofMicrofluidicDevice YiZhang1,2,ChunxiongLuo1,KeZou1,2,ZhengweiXie1,OnnBrandman3,QiOuyang1,4*,HaoLi2* 1The State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, Center for Quantitative Biology, and School of Physics, Peking University, China, 2DepartmentofBiochemistryandBiophysicsandCaliforniaInstituteforQuantitativeBiosciences,UniversityofCaliforniaSanFrancisco,SanFrancisco,California,United States of America, 3Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California, United States of America, 4DepartmentofPhysics,Peking-TsinghuaCenterforLifeSciences,PekingUniversity,Beijing,China Abstract Amajorlimitationtoyeastagingstudyhasbeentheinabilitytotrackmothercellsandobservemolecularmarkersduring theagingprocess.Thetraditionallifespanassayreliesonmanualmicro-manipulationtoremovedaughtercellsfromthe mother, which is laborious, time consuming, and does not allow long term tracking with high resolution microscopy. Recently, wehavedevelopedamicrofluidic system capable of retaining mothercells in themicrofluidic chambers while removingdaughtercellsautomatically,makingitpossibletoobservefluorescentreportersinsinglecellsthroughouttheir lifespan.Herewereportthedevelopmentofanewgenerationofmicrofluidicdevicethatovercomesseverallimitationsof the previous system, making it easier to fabricate and operate, and allowing functions not possible with the previous design. The basic unit of the device consists of microfluidic channels with pensile columns that can physically trap the mother cells while allowing the removal of daughter cells automatically by the flow of the fresh media. The whole microfluidic device contains multiple independent units operating in parallel, allowing simultaneous analysis of multiple strains. Using this system, we have reproduced the lifespan curves for the known long and short-lived mutants, demonstrating the po