Drop-on-Demand Single Cell Isolation and Total RNA Analysis 英文参考文献.docVIP

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Drop-on-Demand Single Cell Isolation and Total RNA Analysis 英文参考文献.doc

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Drop-on-Demand Single Cell Isolation and Total RNA Analysis 英文参考文献

Drop-on-DemandSingleCellIsolationandTotalRNA Analysis SangjunMoon1,Yun-GonKim1,LingshengDong2,MichaelLombardi2,EdwardHaeggstrom3,RoderickV. Jensen4,Li-LiHsiao2*,UtkanDemirci1,2,5* 1DemirciBio-Acoustic-MEMSinMedicine(BAMM)Laboratory,CenterforBioengineering,BrighamandWomen’sHospital,HarvardMedicalSchool,Boston,Massachusetts, United States of America, 2Renal Division, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts, United States of America, 3Electronics ResearchLaboratory,DepartmentofPhysics,UniversityofHelsinki,Helsinki,Finland,4DepartmentofBiologicalSciences,VirginiaTech,Blacksburg,Virginia,UnitedStates ofAmerica,5Harvard-MITDivisionofHealthSciencesandTechnology,MassachusettsInstituteofTechnology,Cambridge,Massachusetts,UnitedStatesofAmerica Abstract Technologiesthatrapidlyisolateviablesinglecellsfromheterogeneoussolutionshavesignificantlycontributedtothefield ofmedicalgenomics.Challenges remainbothtoenableefficientextraction,isolationandpatterningofsinglecells from heterogeneoussolutionsaswellastokeepthemaliveduringtheprocessduetoalimiteddegreeofcontroloversinglecell manipulation.Here,wepresentamicrodropletbasedmethodtoisolateandpatternsinglecellsfromheterogeneouscell suspensions(10%targetcellmixture),preserveviabilityoftheextractedcells(97.060.8%),andobtaingenomicinformation from isolated cells compared to the non-patterned controls. The cell encapsulation process is both experimentally and theoreticallyanalyzed.Usingtheisolatedcells,weidentified11stemcellmarkersamong1000genesandcomparetothe controls.Thisautomatedplatformenablinghigh-throughputcellmanipulationforsubsequentgenomicanalysisemploys fewerhandlingstepscomparedtoexistingmethods. Citation: Moon S, Kim Y-G, Dong L, Lombardi M, Haeggstrom E, et al. (2011) Drop-on-Demand Single Cell Isolation and Total RNA Analysis. PLoS ONE 6(3): e17455.doi:10.1371/journal.pone.0017455 Editor:DimasCovas,UniversityofSaoPaulo-USP,Brazil ReceivedSeptember13,2010;AcceptedFebruary3,2011;Publish