自動(dòng)血液分析儀課件

自動(dòng)血液分析儀---SYSMEXXE-2100COULTERGen.S病理檢驗(yàn)部一般檢驗(yàn)科簡(jiǎn)介更多的分析項(xiàng)目C:WBC,RBC,HGB,HCT,MCV,MCH,MCHC,PLT-ORDW-SD,RDW-CV,PDW,MPV,PCT,P-LCRD:NEUT%,LYMPH%,MONO%,EO%,BASO%NEUT#,LYMPH#,MONO#,EO#,BASO#N:NRBC%,NRBC#R:RET%,RET#,HFR,MFR,LFR,IRF,PLT-I32個(gè)分析項(xiàng)目,分別歸為4類32分析項(xiàng)目選擇選擇不同的分析方式1.C2.C+D3.C+N4.C+D+N5.C+R6.C+D+R7.C+D+N+RC:CBCD:DCN:NRBCR:RETIC操作更方便使用薄膜式按鍵即可完成所有例行作業(yè)關(guān)機(jī)聯(lián)機(jī)求助多功能鍵Sampler上機(jī)輸入Manual上機(jī)輸入光標(biāo)IPU檢體量(自動(dòng)穿刺)(開蓋分析)全新的偵測(cè)管路及分析原理ParametersPrincipleWBC,BASONEUT&BASO,LYMPH,MONO,EONRBCRET,IRF,PLT-OFlowCytometryusingSemiconductorlaserRBC,HCT,MCV,PLT-IHydrodynamicfocusingHGBSLS-HbHPC&ImmatureCellRF/DCDetectorWBC/BasoDIFFNRBCRETRBC/PLTHGBIMI四種偵測(cè)原理,七個(gè)偵測(cè)管路染料染料的成份

Polymethinor/andOxazine染色標(biāo)的

Polymethine:DNA+RNAOxazine:DNA主要分析項(xiàng)目原理介紹RBCHgBPLTWBCDIFFIMINRBCRETRBC分析流程BloodCELLPACKRBC/PLTDetectorBlockSRV2,000μL-Blood:4.0μL-Reagent:1,996μLRBCsamplechamberCELLSHEATHImpedance(DCMethod)ExternalElectrodeApertureInternalElectrode電位=阻抗x電流當(dāng)電流固定時(shí),阻抗與電位成正比血球占去相等體積的稀釋液空間造成阻抗的改變HydrodynamicFocusing原理前后高壓導(dǎo)流導(dǎo)正血球排列及方向流體動(dòng)力聚焦Hb的偵測(cè)偵測(cè)原理:SLSMethod/Spectrophotometer偵測(cè)位置:HbChannel提供項(xiàng)目:HbSLS試藥反應(yīng)SLS厭水端與球蛋白結(jié)合立體結(jié)構(gòu)的改變氧化:Fe2+-->Fe3+SLS親水端與Fe3+鍵結(jié)-->SLS-HbFe2+Fe3+Fe3+SLSSLS-Hb優(yōu)點(diǎn)1.SLS(SodiumLauryalSulfate)是一種Detergent&Surfactant。2.與標(biāo)準(zhǔn)氰化物法相關(guān)性高(R2=0.999)3.Cyanidefreereagent；無(wú)生物毒性。4.Formadehydefreereagent；不刺激呼吸系統(tǒng)。5.沒有Oxyhemoglobinmethod無(wú)法氧化Methemoglobin

的缺點(diǎn)。Impedance/HydrodynamicFocusing原理PLT-I的HistogramFCM/Laser/Fluorescence原理FlowCellCELLPACKCELLPACKSampleSemiconductorLaserLaserbeam:633nmFCM/Laser/Fluorescence原理側(cè)散亂光RNA,DNA的含量細(xì)胞內(nèi)顆粒,結(jié)構(gòu),復(fù)雜性細(xì)胞的大小前散亂光螢光激光光DichroicmirrorPLT-oScattergramForwardScatter(大小)Fluorescence(DNA,RNA)GiantPLTFragmentedRBCSmallPLTXE-2100利用RNA含量的差異,PLT含RNA,成熟RBC不含以光學(xué)原理區(qū)分RBC與PLT,使其成為不連續(xù)的分布WBC,DIFF偵測(cè)偵測(cè)原理:FCM/LaserScatterLight/Fluorescence-偵測(cè)前散亂光,側(cè)散亂光

-偵測(cè)DNA,RNA含量偵測(cè)位置:1.WBC/BasoChannel2.DIFFChannel提供項(xiàng)目:WBC#,Nut#&%,Lym#&%,Mono#&%Eo#&%,Ba#&%DIFF試藥反應(yīng)-normalcellLymMonoNeut+BasoEo1.針對(duì)Eosinophil以O(shè)rganicacid處理,加強(qiáng)SideScatterLight,以增加Eo偵測(cè)敏感度2.中性試劑,含兩種接口活性劑,能將RBC溶解,而白血球則只有輕微的皺縮,抗溶解RBC沒有RNA,

也沒有顆粒,而儀器忽略“FSc(大小)＂的偵測(cè),

因此沒有紅血球破壞不全的現(xiàn)象,不會(huì)干擾分類反應(yīng)前反應(yīng)后SSc(內(nèi)容物)Flu(RNA)DIFFScattergram(DNA,RNA)(內(nèi)容物結(jié)構(gòu))DIFF試藥反應(yīng)-abnormalcell反應(yīng)前反應(yīng)后ImmatureWBC含有較多的RNA成份,因此螢光反應(yīng)較成熟的WBC強(qiáng)DIFFScattergram(abnpattern)SideScatter(Internalstructure)BandMetaMyeloProMyeloBlastAty-LymFluorescence(RNA,DNA)DIFFScattergaramNORMALABNORMALImmatureWBC偵測(cè)XE-2100提供兩種分析原理偵測(cè)ImmatureWBC

偵測(cè)原理1:FCM/LaserScatterLight/Fluorescence

以“RNA含量＂,“細(xì)胞內(nèi)結(jié)構(gòu)信息＂偵測(cè)偵測(cè)位置:DIFFChannel偵測(cè)原理2:RF/DC

以“細(xì)胞膜脂質(zhì)含量＂,“細(xì)胞核密度＂偵測(cè)偵測(cè)位置:IMIChannel提供項(xiàng)目:Blast?,IG?,LS?,Aty-Ly?,HPC#&%,IG#&%(HPC:HaematopoeticProgenitorCell,造血前驅(qū)細(xì)胞,相當(dāng)于CD34+Cell

也就是PBSC,周邊血液干細(xì)胞)ImmatureWBC偵測(cè)原理-1SideScatter(Internalstructure)BandMetaMyeloProMyeloBlastAty-LymFluorescence(RNA,DNA)以“RNA含量＂,“細(xì)胞內(nèi)結(jié)構(gòu)信息＂偵測(cè)ImmatureWBC偵測(cè)原理-2RF:RadiofrequencyDC:DirectCurrent(Impedance)細(xì)胞的大小細(xì)胞核密度IMI試藥反應(yīng)(反應(yīng)前)IMI試藥反應(yīng)反應(yīng)后,成熟的WBC裸核化,ImmatureWBC形態(tài)輕微皺縮IMI試藥反應(yīng)成熟白血球不成熟白血球IMIScattergram細(xì)胞在成熟的過(guò)程中最大的差異在于核密度的變化利用RF來(lái)偵測(cè)細(xì)胞核的密度可以很敏感的將Blast,IG,band等分開RFDCBandMyelocyteMetamyelocyteBlastPromyelocyte以“細(xì)胞膜脂質(zhì)含量＂,“細(xì)胞核密度＂偵測(cè)AbnormalCell分布BandMetaMyeloProMyeloBlastAty-LymFluorescence(RNA,DNA)SideScatter(Internalstructure)PLTClumpImmatureGranBlastHPCLeftshiftRF(細(xì)胞核的密度)DC(大小)ABA可偵測(cè)Lymphoblast/MyeloblastB可偵測(cè)MyeloblastA+B+MyeloblastA+B-LymphoblastXE-2100提供Blasts?以及L-Blast?兩種FlagIMIDIFFnormalLymQ-FlagQ-Flag為儀器對(duì)SuspactMessage“Possibility＂的Index可用在儀器與手工鏡檢比對(duì)時(shí),調(diào)整敏感度畫面中數(shù)字為Q-FlagIndex,代表Possibility定量NRBC的偵測(cè)偵測(cè)原理:FCM/LaserScatterLight/Fluorescence-偵測(cè)前散亂光/側(cè)散亂光

-偵測(cè)DNA,RNA含量偵測(cè)位置:NRBCChannel提供項(xiàng)目:NRBC#,NRBC%(NRBC/100WBC)NRBC試藥反應(yīng)反應(yīng)前反應(yīng)后特殊酸性試劑,含水楊酸成份,能加強(qiáng)RBC的溶解,而WBC則輕微的皺縮,同時(shí)作核酸的染色NRBC試藥反應(yīng)WBCNRBCRBCHighLevelFluorescenceSize:BigLowLevelFluorescenceSize:SmallWeakFluorescenceSize:MicroLysing(WBCmembraneisweaklydamaged)StainingRNADNANRBCScattergram(DNA,RNA)(大小)“Achangeoccursinelectricalresistancebetweentwoelectrodeswhenanonconductiveparticlesuchasabloodcellpassesbetweenthem＂ -WallaceCoulter,1947TheCoulterPrinciple“ScienceServingHumanity.＂AcTAcT5PDHmXSTKSGenSLH755AcT5PDALCountingtheCellsCellsaredilutedinanelectricallyconductivefluid(0.085%saltwater).Electrodesareseparatedbyasmallholethroughwhichthecurrentflows.Asthefluidandcellsarepulledthroughthehole,thecelltemporarilyblockstheflowofcurrentandcreatescountablepulses.SizingtheCellsThelargerthecell,themoreelectricalcurrentitblocks.Theheightofthepulseisequaltothevolumeofthecell.TheCoultermethodcountsandsizescellsbydetectingandmeasuringchangesinelectricalresistancewhenaparticle(suchasacell)inaconductiveliquidgoesthroughasmallaperture.#ofPulseParticlecountSizeofPulseCellVolumeCBCAnalysis-ImpedancePrincipleSensingZoneRedBloodCellAredcellpassesthroughRBCapertureTheCoulterprincipleOscilloscopeOscilloscopeOscilloscopeSensingZoneNeutrophilAWhitecellpassesthroughWBCapertureTheCoulterprincipleOscilloscope20fl2flBaselineRBCofvarioussizesPlateletsApplyingThresholdstoseparatePLTfromRBCHydrodynamicFocusing檢體不會(huì)與Sheathfluid混合確定檢體通過(guò)偵測(cè)孔正中央防止回流HydrodynamicFocusingwithDiluentSheath8psiCellsApertureSensingZoneRBCspushedawaybydiluentOscilloscopescreenRBCPLT+-DiluentstreamSystemwithsweepflow

掃流系統(tǒng)SweepFlowPulsetobeeditedDiluentstreamPulseediting(脈沖修正)NoncentralcellflowHemoglobin(HGB)國(guó)際標(biāo)準(zhǔn)法:氰化高鐵血色素法原理溶解紅血球?qū)⒍r(jià)鐵氧化為三價(jià)鐵之高鐵血色素高鐵血色素與KCN結(jié)合為氧化高鐵血色素

540nm比色優(yōu)點(diǎn)專一性佳缺點(diǎn)

KCN具毒性且造成環(huán)境污染

光學(xué)比色法原理Cyanide-freereagentBeer＇sLaw測(cè)定血紅素稀釋液的吸光度變化吸光度波長(zhǎng)為540nm血紅素復(fù)合物為吸收光線的主要成分MCV直接測(cè)定法將雙凹盤狀紅血球球狀化直接偵測(cè)紅血球通過(guò)時(shí)發(fā)生電阻波形之高度推算之體積避免deformedRBCMCV偏低較傳統(tǒng)計(jì)算法更能看出紅血球?qū)嶋H大小PackedCellVolume(PCV;Hematocrit)WintrobetubemethodMicrohematocritmethodAutomaticmethodRedcelldistributionwidth(RDW)反應(yīng)紅血球大小分布及差異性PriceJohns曲線圖直徑測(cè)定法自動(dòng)血球計(jì)算法Anisocytosis

RDW-CV=×100%MCV

SD血小板測(cè)定方法

人工計(jì)數(shù)法電子阻抗法光學(xué)測(cè)定法免疫血小板電子阻抗法

自動(dòng)血球分析儀最常見的原理以Hydrodynamicfocusingflow,測(cè)單一方向大小

將特定大小范圍內(nèi)的物質(zhì)定為血小板白血球計(jì)數(shù)及分類測(cè)定需將紅血球溶解以阻抗法測(cè)數(shù)目分類使用ScatterLight偵測(cè)

(3-partdifferential)CoulterWBCHistogram

(3-partdifferential)LymMonoEosNeutBaso

WBC50100200300400fLLyEoNeRBCErythroLyseStabliLyseNearNativeFlowcyometryVCSTechnologyDirect5-partDifferentialCountTechnologyRecap最佳血球大小偵測(cè)方法偵測(cè)0~3