macs磁性分選與facs流式細胞優(yōu)勢比較

MACS分選與FACS分選10823分鐘，分選1094個小時。而且流式分選需要調(diào)整儀給機器調(diào)節(jié)帶來，而獲取條件不合適，必定會直接影響分選結(jié)果。而MACS分選速度107細胞/1095分鐘時間，大大地提高了工作效率。同時MACS分操作簡單：流式分選需要專門的有豐富經(jīng)驗的操作人員。而MACS分選操作簡單，分選后細胞的：在流式細胞儀分選中，細胞長時間處于高壓液流包裹狀態(tài)，加上分選時間長，導(dǎo)致分選后得到的細胞狀態(tài)不好。而MACS分選，由于分選速度快，所需時間短，并且磁珠只有50nm，對細胞無毒性，能最大程度的保留細胞的。裁判員均為同一人的現(xiàn)象，無法真正保證分選結(jié)果的可靠性。而MACS分選后，用流式細回收率：在流式分選中，流式進樣和獲取管道決定了流式分選最高只能獲取80％的總體細胞，加上調(diào)節(jié)儀器條件也會造成細胞損失，因此回收率低。而MACS分選采用批處理方式，能盡可能回收目的細胞?；厥章试?0%以上。分選成本上：流式分選需要的試劑是抗體，每支抗體的平均價格在2000能標記108細胞。而MACS分選需要的磁珠，平均價格在8500左右，但是能分選109細胞。因此MACS分選的成本要遠遠低于FACS分選。（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."分選MACS&More,ApplicationofuMACSStreptavidinMicroBeadsfortheysisofHIV-1directlyfrompatientInthisreport,wenowdescribemarkedimprovementsintheimmunomagneticcaptureprotocolthatpermitsysisofvirionsdirectlyfrompatientsplasmawithoutpriorprocessing.Theimprovedabilityviralcapturetechnologywithoutextensivesampleprocessing,duetotheuniquepropertiesofmMACSMicroBeads,canadvancethistechniqueintonewapplications.Byeliminatingtheneedtoremoveinhibitors(antibodies,acuteserumproteinsetc.)frombiologicfluidsbyprocessing,thiswillallowtheapproachdesscribedheretobeutilizedforbodilyfluidsforwhichextensiveprocessingwouldnotbefeasible(breastmilk,semen,etc.).2:關(guān)于MACS分選后細胞活性的文獻（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分選后細胞功能的文獻(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分選后電鏡檢查Leuk.CharacterizationofCD34+HumanHematopoieticProgentitorCellsfromthePeripheralBlood:Enzyme-,Carbohydrate-andImmunocytochemistry,Morphometry,andUltrastructure.CD34+cellswereisolatedusingtheCD34+IsolationKitandCD34+cellswereisolatedusingtheCD34+IsolationKitandMiniMACS(Purity90-95%,Reco