本科论文细胞工程3000字翻译.docx

本科论文细胞工程3000字翻译.docx

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本科论文细胞工程3000字翻译

本科毕业设计（论文）

英文文献翻译

学院名称：

电气信息工程学院

专业：

电子信息工程

班级：

14电子1B

学号：

201496111

姓名：

李鹏

指导教师姓名：

指导教师职称：

二〇一六年六月

Part1：

cellengineering

In2007,Lietal.[94]proposedtheirmicrofluidicdeviceforsinglecellanalysis.Thechipwasfabricatedbystandard1-photomaskwithlowcostmethod.Thedeviceconsistsfourreservoirs,fourchannelsandoneopenregionwhichcontainacellretentionchamber.Reservoironewasusedforcellintroductionandwashing,whereasreservoirtwowasusedforreagentdelivery.Reservoirthreeandfourwereusedaswastereservoirs.Byusingthischip,theyquantifieddynamicCa2+mobilizationofasinglecardiomyocyteduringitsspontaneouscontraction.Also,theymonitoredsuccessfullydynamicresponsesfromvariousexternalstimulationsuchasdaunorubicin（cardiotoxicchemotherapeuticdrug）,caffeine,andisoliquiritigenin（herbalanticancer）.TheirresultsalsoprovethatanticancerdrugshavelesseffectontheCa2+ofthecardiomyocytes.Thisdevicehasquantifiedthecellularresponseofsinglecardiomyocytes,discoveryofheartdiseasesdrugandcardiotoxicitytesting.

In2010,Mellorsetal.[95]proposedanelectrophoreticandelectrosprayionizationbasedmicrofluidicdeviceforsinglecellanalysis.Thedevicewasfabricatedoncorningborosilicateglasssubstratebyusingstandardphotolithographyandwetchemicaletchingtechnique.Figure12showstheschematicofthemicrofluidicdevice,whereAwasacellloadingreservoirandBwasbufferloading,whichintersectswiththeseparationchannel

Thisintersectionzonewasacelllysiszone.CSwasanelectro-osmoticpumpwhichwasconnectedwithanelectrophoreticseparationchannelandelectrosprayorifice.Cellscanflowthroughhydrodynamicallyorelectricallytotheintersectionzone,wherecellswereelectricallylysed.Then,cellscanmigratetoelectrosprayorificethroughtheseparationchannelwherecellselectrosprayionizationoccurred.Thisdevicesuccessfullylysedhumanerythrocyteswithreal-timeelectrophoreticseparation.Thehemegroup,αandβsubunitsofhemoglobinweredetectedfromerythrocyteswhencellswerecontinuouslyflowedthroughthedevice.Thisdevicecananalyze12c/m.

Localizedsinglecellmembraneelectroporationcanprovidebettercelltransfectionwithmicro/nanofluidicdevicescomparedtosinglecellelectroporation（SCEP）orbulkelectroporation（BEP）.Becauseofmicro/nanoscaleelectrodedimensionanddistancebetweentwoelectrodeswereverysmall,asaresult,electricfieldcanintenseinaverysmallregionofthecellmembranecomparedtosinglecelldimension.Thus,thelocalareaofthesinglecellcanbeaffectedbyastrongelectricfield,whereasotherareaswillbeunaffected.Duetotheeffectsofsmallareasofthewholesinglecell,highcellviabilityandhightransfectionratecanbeachievedcomparedtosinglecellelectroporation.However,Boukanyetal.showlocalizedsinglecellelectroporationbyusingnano-channelbasediontransportationusingelectrophoresismethodwithtwolargeelectrodes[30].Byfabricatingmicro/nanoelectrodeswithamicro/nanoscaleelectrodegap,thisdevicecanprovidesomepromisingparameterssuchaslowvoltageandpowerrequirement,lowertoxiceffectduetonegligibleiongeneration,smallsamplevolumeandnegligibleheatgeneration.Theseparametersareessentialtoachievehightransfectionrateandhighcellviability.Thus,microfluidicbasedLSCMEPprocesscanprovideabetterunderstandingtoanalyzeintracellularcytosoliccompoundscomparedtoSCEPorthebulkelectroporationprocess.Nawarathnaetal.,demonstratedtheAFMbasedLSCMEPprocess.Figure13showslocalizedelectroporationofasinglecellusingatomicforcemicroscopy（AFM）technique[29].Forthisexperiment,theymodifiedAFMtiptoactasanano-electrodetomakeanintensehighelectricfieldnearthelocalizedareaofthesinglecellmembrane.AborondopedsiliconAFMtips（σ=0.001Ωcm,k=1.5N/m）wasusedforLSCMEPprocess.Beforeelectroporation,thetipwasgrownwith20nmSiO2layerandfinallythisoxidizedtipwassectioneduntilbaresiliconwasexposedbyfocusedionbeam（FIB）technique.Asaresult,asmallerareaofbaresiliconcancauseanintensehighelectricfieldonasinglecellmembrane.Theyhavereducedthisbaresiliconareadownto0.5μmindiameter,whichwasconcentratedwithanintenseelectricfieldon10μmdiameterofratfibroblastcell.Figure13a–hshowstheresultsofLSCMEPtechniqueusingAFMtipforelectroporationprocessandFigure13idemonstratedtheAFMtip,whichwaspositionedontopofthesinglecellforlocalizedsinglecellmembraneelectroporation（LSCMEP）process.Tomakeanintensehighelectricfield,1Vppwith0.5Hzpulsewasusedtotransfectratfibroblastcells.Thetransfectionofsinglecellwascompletedwithin10s.Thisdevicecanperformhighlylocalizedelectroporationofasiglecellwithconcentricelectricfieldonlocalareaofsinglecellmembrane.Theexperimentcanbeperformedinafriendlyenvironmentsuchascellculturedishes,etc.

Inrecentyears,Boukanyetal.[30]showednanochannelbasedlocalizedsinglecellelectroporationwithapreciseamountofbiomoleculesdelivery.Inthisdevice,theypositionedasinglecellinonemicrochannelbyopticaltweezersandtransfectionagentwasloadedtoanothermicrochannel.Thesetwomicrochannelswereconnectedbyonenanochannel.Toapplyaveryhighelectricfieldinbetweentwomicrochannels,atransfectionagentwasdeliveredthroughthenanochannelusinganelectrophoreticallydrivenprocessandfinallydrugsweredeliveredinsideasinglecellthroughaverysmallareaofthecellmembrane.In2012,Chenetal.demonstratedanotherlocalizedsinglecellmembraneelectroporationusimgITOmicroelectrodebasedtransparentchip[27].Figure14showsmicrofluidiclocalizedsinglecellmembraneelectroporationdevice.TheydepositedITOfilmsonacoveredglasssubstrateandpatterenditbystandardlithographicprocesstoformasITOlines.Afterthat,athinSiO2layerwasdepositedasapassivationlayerbyplasmaenhancedchemicalvapordeposition（PECVD）technique.ThefinalITOlineswerecutbythefocusedionbeam（FIB）technique.Thegapbetweentwoelectrodeswere1μmandwidthofeachelectrodewas2μm.Whensinglecellwasstronglyattachedinbetweentwoelectrodesgap,theelectricfieldwasintensedinonlya1μmgapareaonsinglecellmembrane.Asaresult,theydemonstratedlocalizedsinglecellmembraneelectroporationwithmicrofluidicdevice.Figure14ashowslocalizedelectroporationprocessbetweentwomicro-electrodesandFigure14bshowsmultiplenumberofelectrodesforLSCMEPprocess.Figure14canddshowstheopticalmicroscopeimageofpatterenedITOmicroelectrodesandscanningelectronmicroscope（SEM）imageofITOmicroelectrodsewithmicro-channel.Accordingtotheirresults,theyachived0.93μmelectroporationregionwith60%cellviabilityfor8Vpp20mspulseapplication.Toreducethegapbetweentwoelectrodes,ahightransfectionratecanbeachivedbythistechnique.Thisdevicenotonlycontroltherecoveryofcellmembranes（reversibleelectroporation）withoutcelldamagebutalsoitprovidesclearopticalviewbyusinganinvertedmicroscope（ITObasedtransparentchip）.

Recently,anotherLSCMEPbaseddevicewasproposedbyJokilaaksoetal.[77]forsinglecelllysis.Theyreportedasiliconnanowireandnanoribbonbasedbiologicalfieldeffecttransistorforsinglecellpositioningandlysismechanism.Figure15ashowsthecrosssectionalviewandelectricconnectionwithPDMSabovethedeviceandFigure15bshowsanarrayofthetransistorswithbothnanowiresandnanoribons.Topositionthesinglecellonthisdevice,theyusedprogrammablemagneticfieldformagneticmanipulationof7.9μmCOOHmodifiedCOMPELmagneticmicrosphere.Afterpositioningthesinglecell（HT-29）ontopofthetransistor,cellswereadheredfor30minpriortoelectroporationexperiment.Theappliedelectricfieldwas600–900mVpp（peaktopeak）at10MHzfor2mspulse.Thiselectricfieldwasconnectedwithashortedsourceanddraininoneterminalandanotherterminalconnectedonthegateofthedevice.Theelectricfieldintensitywasfringinginnature,whichaffectedthecellmembraneintegrityleadingtocelllysis.Thisdevicecanperformsinglecelllysiswhichispotentiallyapplicabletomedicaldiagnosticsandbiologicalcellstudies.

Insummary,thisarticledescribesthedetailsaboutbulkelectroporation（BEP）,singlecellelectroporation（SCEP）,andlocalizedsinglecellmembraneelectroporation（LSCMEP）byusingmicro/nanofluidicdeviceswiththeiradvantagesanddisadvantages.Alloftheseprocessescandeliverdrugs,DNA,RNA,oligonucleotides,proteins,etc.However,toanalyzecelltocellbehaviorwiththeirorganellesandintracellularbiochemicaleffect,singlecellanalysismustbeexecuted.Micro/nanofluidicdevicesarethepotentialcandidatestoanalyzesinglecells,becauseoftheirdimensionreductiontothedimensionofsinglecelllevel.Thesedevicesprovideeasyperformancesuchascellhandling,lowerpowerconsumption,lowtoxicity,smallsamplevolume,lowercontaminationrate,highcellviability,andhightransfectionratewhencomparedtoconventionalelectroporation.Toreducetheelectrodeareaandgapbetweentwoelectrodesbyusingmicro/nanofluidicdevices,selectiveandlocalizeddrugdeliveryispossible.Thisnewapproachiscalledlocalizedsinglecellmembraneelectroporation（LSCMEP）.However,untilnowthistechniqueisinthedevelopmentstage.Inthefuture,theLSCMEPprocesscanprovideselectiveandspecificsinglecellmanipulationfrommillionsofpopulationsofcellstog