macs磁性分选与facs流式细胞优势比较

MACS分选与FACS分选10823分钟，分选1094个小时。而且流式分选需要调整仪给机器调节带来，而获取条件不合适，必定会直接影响分选结果。而MACS分选速度107细胞/1095分钟时间，大大地提高了工作效率。同时MACS分操作简单：流式分选需要专门的有丰富经验的操作人员。而MACS分选操作简单，分选后细胞的：在流式细胞仪分选中，细胞长时间处于高压液流包裹状态，加上分选时间长，导致分选后得到的细胞状态不好。而MACS分选，由于分选速度快，所需时间短，并且磁珠只有50nm，对细胞无毒性，能最大程度的保留细胞的。裁判员均为同一人的现象，无法真正保证分选结果的可靠性。而MACS分选后，用流式细回收率：在流式分选中，流式进样和获取管道决定了流式分选最高只能获取80％的总体细胞，加上调节仪器条件也会造成细胞损失，因此回收率低。而MACS分选采用批处理方式，能尽可能回收目的细胞。回收率在90%以上。分选成本上：流式分选需要的试剂是抗体，每支抗体的平均价格在2000能标记108细胞。而MACS分选需要的磁珠，平均价格在8500左右，但是能分选109细胞。因此MACS分选的成本要远远低于FACS分选。（1.CalciumIonophore-TreatedPeripheralBloodMonocytesandDendriticCellsRapidlyDisplayCharacteristicsofActivatedDendriticCells.isolationofmonocytesandimmaturedendriticcells(iDC) fromelutriatedleukapheresatesusingCD14andCD33MicroBeads,VS+columnsandMidiMACSorVarioMACS.AuthorsclaimthatFACSsortingofthecellsimpairedtheirresponsivenesstoionophortreatmentandthustheyusedMACSseparationtocircumventthis.ComparisonofPurityandEnrientofCD34+CellsfromBoneMarrow,UmbilicalCordandPeripheralBlood(PrimedforAphresis)UsingFiveSeparationSystems.EnrientofCD34+cellsusingCD34KitandTheauthorshaveusedFACSsortingandfourofthelabotary-scaledevices(Cellector,Dynabeads,CEPRATE,MiniMACS)whicharebasedonpositiveselectionbyimmunoadsorptiontocompareenrientofCD34+cellsfromdiffrentsamples.Theisolatedcellswereassessedforpurity,yieldandcolonyformingcell(CFC)enrient.MACSshowedcomparablepuritytoFACSandCellectorFlasksBUToutofalltechniquesMACShadhighestyieldandcolonyformingcell-enrient.（3）Proc.Natl.Acad.Sci.USA，1994，91：1323－1327 binationinNormalIgA1+BLymphocytes.HumanBcellsexpressingall-surfaceIgA1werepurifiedfromperipheralblood.AfterFicollgradientcentrifugation,washingtoremoveplaetsandleucinemethylesterlysistodepletemonocytes,lymphocyteswerelabeledwithbiotinylatedanti-IgA1antibody,StreptavidinMicroBeadsandStreptavidin-PE.IgA1+wereenrichedtofrequenciesofupto80%withrecoveriesofupto80%.MACSprovedmuchmoreefficientthanFACSintermsofpurityandrecovery.Theshortprocessingtime(<1h)andlowphysicalforcesrequiredforMACS,comparedtoFACS,alsoresultedinhighercellviability.Tofurtherpurifythecellstohomogenityformolecularysis,MACS-enrichedcellswereprocessedfurtherbyFACS(finalpurity>97%).TheBiologyandApplicationofHumanBoneMarrowStromalCellRecentdataconcerningtheimmunophenotypeandfunctionalcharacteristicsofprecursorcellsfrommarrowstromaltissuewerereviewed.Theauthorscompareddifferentisolationstrategiesofstromalmarrowcells,e.g.FACS,DynalandMACS,andtheypraisedMACStechnologyasthebest:MACSprovidesaparticularlyeffectiveisolationtechniquethatcanbeemployedusingBMmononuclearcellswithoutanypreenrientsteps,reproduciblyproducesexcellentyieldsofSPC,andcanreadilyprocesshighcellStromalcellswereisolatedwithStro1-abandindirectsystem.ForexplicitprotocolseeGronthosandSimmons[No.149].Am.J.Pathol.1996,150:1021-ExpressionPatternandCellularOriginofCytokinesintheNormalandToxoplasmagondii-infectedmurinebrainMACSisolationofCNS-derivedleukocytes:usingeitherdirectCD4orCD8MicroBeadsorindirectlywithF4/80(recognizesmacrophagesandmicroglia)+goatanti-ratMicroBeadsTheauthorsinvestigatedtheintracerebralcytokineproductioninnormalandToxoplasmagondiiinfectedusingimmunohistochemistry,insituhybridization,flowcytometryandRT-PCR.Therefore,theyisolatedCD4+,CD8+Tcellsandmacrophages/microgliacells(F4/80bright/F4/80dim)usingMACStechniquefrombrainofnormalandinfectedmice.TheyfirsttriedtoisolatethesecellswithFACsorting,however,noviablecellscouldberetrievedfrombrainparenchyma...(probably)basedonhighlevelofapoptoticleukocytes...Therefore,amoregentlemethodwhichisMACSwasestablished.Purityoftheisolatedcellpopulationswas>97%,recoveryafter2columnpassageswas~25%.DuetolownumberofCD4+/CD8+TCsinnormalbrain,thesecellscouldonlybeisolatedfrominfectedmice.TheMACSisolationallowedafurthersemitativeRT-PCRforcomparisonofcytokinemRNAindifferentcells.（6）Int.J.Radiat.ImproveddeterminationofvarianterythrocytesattheglycophorinA(GPA)locusandvariantfrequencyinpatientstreatedwithradioiodineforthyroidcancer.ysisoftheGPAgeneasameansforthedeterminationofmutationsduetoe.gradiationsincemutationsaquiredbystemcellsareretainedinerythrocytes.BR6assayissofartheonlyassayusedfortheysisoftheappearanceofredbloodcellvariants.Normallycellsarestainedandysedflowcytometrically.ButtoofewcellscanbeysedthiswaythereforepreenrientwithMACS.HighlyabundantgenesinthetranscriptosomeofhumanandbaboonCD34antigen-positivebonemarrowcells.Usedbiotinylatedanti-CD34antibodies12.8(baboon)orQBEND/10(human)andstreptavidin(SA)beadstopullouthumanandbaboonhematopoeiticstemcellsAuthorsfavorablycomparedMACStoflow"ExpressionstudiesoncDNAarraysrequireafairlylargenumberofcellstoisolateanappropriateamountofRNAforprobepreparation.Becauseofthisconstraint,itwasnecessarytopurifytheCD34+cellsbyimmunomagneticcolumnsratherthanFACS,whichwouldrequireprolongedsorting.ThestressimposedbytheprolongedsortingtimerequiredtopreparethisnumberofcellscandramaticallyreducecellviabilityandyieldofCD34+cellsandmayaltertheirgeneexpressionprofile."分选MACS&More,ApplicationofuMACSStreptavidinMicroBeadsfortheysisofHIV-1directlyfrompatientInthisreport,wenowdescribemarkedimprovementsintheimmunomagneticcaptureprotocolthatpermitsysisofvirionsdirectlyfrompatientsplasmawithoutpriorprocessing.Theimprovedabilityviralcapturetechnologywithoutextensivesampleprocessing,duetotheuniquepropertiesofmMACSMicroBeads,canadvancethistechniqueintonewapplications.Byeliminatingtheneedtoremoveinhibitors(antibodies,acuteserumproteinsetc.)frombiologicfluidsbyprocessing,thiswillallowtheapproachdesscribedheretobeutilizedforbodilyfluidsforwhichextensiveprocessingwouldnotbefeasible(breastmilk,semen,etc.).2:关于MACS分选后细胞活性的文献（1）Proc.Natl.Acad.Sci.CD8+TLymphocytesofAfricanGreenMonkeysSecreteanImmunodeficiency-SuppressingLymphokine.TheauthorsinvesitgatedtheinfluenceofCD8TlymphocytesonSIVreplicationinCD4TlymphocytesofAfricangreenmonkey(AGM).TheauthorsusedCD8microbeadsfordepletionofCD8+cellsfromPBMC.AdditionallypositiveselectionofCD4andCD8cellsofAGMswasperformedafterdepletionofmonocytesusingCD14PurityofisolatedTcellfractionswas>98%withviabilityof>EffectofGranulocyte-MacrophageColony-StimulatingFactoronEicosanoidProductionbyMononuclearHumanPBMCwereisolatedfromvenousblood,Ficoll-purified,filteredthroughnylonmeshandthenlabeledwithCD14MicroBeadsandMACSseparatedAuthorsexaminedtheeffectsofGM-CSFonarachidonicacidmetabolisminratalveolarmacrophages,peritonealmacrophagesandMACSisolatedhumanperipheralbloodmonocytes.MacrophagesandmonocyteswereculturedandtreatedwithGM-CSFandtheneicosanoidproductionwasmonitored.TheMACSpurifiedmonocyteswere97%pureandvital(determinedbytrypanblueexclusionatisolationandafterovernightincubation).Preparedcellswerenotactivatedtogenerateeicosanoids.3:MACS分选后细胞功能的文献(1)J.Immunol.1996,158:637-TheroleofB7-1andLFA-3incostimulationofCD8+isolationofCD4+andCD8+TCfromhumanPBMCusingdirectbeads-- TCfromPBMCafterFicollisolatedusingdirectbeadspurity>98%andCD8>96%cellsusedininvitroassaysligatione.g.withspecificantibodiesauthorsmentionthatMACSsortedcellscanbedirectlyusedforthese'MACSMicroBeadsarebiodegradable posewhencellsare'Theytypicallydonotactivateisolated influencetheirviability,andnobead entisrequired,positivelyisolatedcellscanbeusedimmediaEur.J.ArePrimedCD4+TLymphocytesDifferentfromUnprimedAuthorsinvestigatedthecontributionofantigen-specificTCfrequencytothegenmerationofprimaryandsecondaryCD4+TCresponsesinvitroandinvivo.TheproliferationandIFN-gproductionafterinvitroprimingandrestimulationofnaiveCD45RB+TC(MACSisolated)wasmeasured.MACSpositiveselectionstrategydidnotinfluencenaiveTCfunction!!4:MACS分选后电镜检查Leuk.CharacterizationofCD34+HumanHematopoieticProgentitorCellsfromthePeripheralBlood:Enzyme-,Carbohydrate-andImmunocytochemistry,Morphometry,andUltrastructure.CD34+cellswereisolatedusingtheCD34+IsolationKitandCD34+cellswereisolatedusingtheCD34+IsolationKitandMiniMACS(Purity90-95%,Reco