PrinciplesofmeasurementBiomedicalengineering測(cè)量原理生物醫(yī)學(xué)工程課件

PrinciplesofmeasurementBiomedicalengineering測(cè)量原理生物醫(yī)學(xué)工程課件DCDetectionMethodThePulseSampleBeakerFirstDilution 1:500(WBC)SecondDilution 1:50.000(RBC)

SampleBeakerDB-1DilutionsforSemiautomaticAnalyserDCDetectionMethodThePulseSDCDetectionMethodThePulseWBCDilution+3DropsQuicklyserforLysingandHGBRBCDilutionDilutionsforSemiautomaticAnalyserDCDetectionMethodThePulseWDCDetectionMethodThePulseExternalElectrodeApertureInternalElectrodeDCDetectionMethodThePulseEDCDetectionMethodTransducerVacuumBloodcellsResistanceConstantCurrent(ca.100V)InternalElectrodeExternalElectrodeApertureSampleBeakerBloodSuspensionDCDetectionMethodTransducerDCDetectionMethodTransducerExternalElectrodeApertureInternalElectrodeVacuumDCDetectionMethodTransducerDCDetectionMethodExternalElectrodeApertureInternalElectrodeU=RxIThePulseDCDetectionMethodExternalApDCDetectionMethodU=RxIExternalElectrodeApertureInternalElectrodeThePulseDCDetectionMethodU=RxIEDCDetectionMethodTime[sec]PulsePictureVolume[fl]Pulses/VolumeBloodsuspensionTransducerStart-SensorStop-SensorManometerCountingTime(definedVolume)forerunafterrunAbsoluteCountingIDCDetectionMethodTime[secTime[sec]PulsePictureVolume[fl]Pulses/VolumeBloodsuspensionTransducerStart-SensorStop-SensorManometerCountingTime(definedVolume)PrerunAfterrunDCDetectionMethodAbsoluteCountingITime[sec]PulsePictureVolumeTime[sec]PulsePictureVolume[fl]Pulses/VolumeBloodsuspensionTransducerStart-SensorStop-SensorManometerCountingTime(definedVolume)PrerunAfterrunDCDetectionMethodAbsoluteCountingITime[sec]PulsePictureVolumeTime[sec]PulsePictureVolume[fl]Pulses/VolumeBloodsuspensionTransducerStart-SensorStop-SensorManometerCountingTime(definedVolume)PrerunAfterrunDCDetectionMethodAbsoluteCountingITime[sec]PulsePictureVolumeTime[sec]PulsePictureVolume[fl]Pulses/VolumeBloodsuspensionTransducerStart-SensorStop-SensorManometerCountingTime(definedVolume)PrerunAfterrrunDCDetectionMethodAbsoluteCountingITime[sec]PulsePictureVolumeDCDetectionMethodTransducerSampleStartStopDefinedVolumeVacuumCounteroffanDisplaySumofPulses/VolumeonPulseProcessingdigitalAbsoluteCountingDCDetectionMethodTransducerDCDetectionMethodTransducerSampleStartStopDefinedVolumeVacuumCounteroffanDisplaySumofPulses/VolumeonPulseProcessingdigitalAbsoluteCountingIISignal-PulsesDisturbingPulsesSignal-PulsesDiscriminatorNottobechangedbyUser:

SampleVolume

DilutionRatio

CountedVolume

ApertureDiameterAutomaticallycheckedbytheinstrument:

CountingTime

Conditionofapertureduringmeasurement

(Noise-Control)Advantages:

Nocalibrationofcounters

AutomaticcheckofapertureDCDetectionMethodTransducerDCDetectionMethodSampleVacuumReferenceSampleforInstrumentCalibrationPulsesperTime

electricalVoltage

Digital-Analog

ConverterAnalog-DigitalConverter

(Digital-Voltmeter)Display: digital Volt(=TargetValueofReferenceSample)AdjustmentofVoltagetoTargetValueofReferenceSamplePulsprocessinganalogSignal-PulsesDisturbingPulseSignal-PulsesDiscriminatorPulsprocessingdigitalMeasurementofParticles

perTimePeriodElementsofUncertainty:

ReferenceSample

StateoftheAperture

VacuumRelativeCountingDCDetectionMethodSampleVacuDCDetectionMethodMeasurementofparticles

inadefinedvolumeCalculationofcell-concentrationwithknownSamplevolumeanddefinedSampledilutionnocalibrationofcountingvaluesMeasurementofparticlespertimeperiodThecountisreceivedindirectlythroughareference-sample.Itistakenforgrantedthatthesampleandthereference-samplehavethesamecharacteristicsandgivethesameresult.CalibrationofcountingvaluesinnecessaryAbsoluteCountingRelativeCountingComparisonofAbsoluteandRelativeCountingDCDetectionMethodMeasuremen1234567891011121314PulseHeightVolume(Time)DCDetectionMethod1234567891011121314PulseHeigh1234567891011121314CumulativeSizeDistributionCurvePulsePicture102030123456789101112131441000123454321DCDetectionMethodHistogram1234567891011121314Cumulative1234567891011121314Histogram41000123454321CumulativeSizeDistributionCurve102030123456789101112131441000123454321DCDetectionMethodHistogram1234567891011121314Histogram41ErythrocyteHistogram25-75fl200-250flDCDetectionMethodHistogramErythrocyteHistogram25-75fl2PlateletHistogram2-6fl12-30flfixedat12flDCDetectionMethodHistogramPlateletHistogram2-6fl12-30DCDetectionMethodHistogramLeukocyte-Histogramfixedat300fl30-60flLDT1T2UDLymphocytesNeutrophilesMonocytesBasophilesEosinophilesDCDetectionMethodHistogramLOperationSequencesHistogramSV2SV3SV1SV4NCNOCNONCCCNONCCNCDiluentInlet(A)(B)BallFoatOpentotheAirorPressurePumpWaste(Vacuum)Allsolenoidvalvesarede-energized.C(Common)andNO(NormallyOpen) solenoidvalveportsareconnected.BallFloatisinthedownposition.“READY”-StatusOperationSequencesHistogramSOperationSequencesHistogramSV2SV3SV1SV4NCNOCNONCCCNONCCNCDiluentInlet(A)(B)BallFloatOpentotheAirWaste(Vacuum)COUNTKEYonSV1andSV4areenergized.CandNCsolenoidvalveportsare connected.Diluentisaspiratedthrough SV4,SV2,Transducer,SV1andSV3.TheBallFloatinthemanometeris pushedupward.Theinnerportionoftransducerand manometerisautomaticallyrinsedto minimizecarry-overeffect.OperationSequencesHistogramSOperationSequencesHistogramCountingSV1andSV4arede-energizedand SV2isenergized.Thedilutedsampleisaspiratedthrough theapertureandtheBallFloatmoves downward.WhentheBallFloatreacheslevelA, thecounterstartscounting.WhentheBallFloatreacheslevelB, thecountingstopsandSV2is de-energized.Thevolumeofaspiratedsampleisthe sameasthemanometer-volume betweenpointsAandB.SV2SV3SV1SV4NCNOCNONCCCNONCCNCDiluentInlet(A)(B)OpentotheAirWaste(Vacuum)OperationSequencesHistogramCOperationSequencesHistogramSV2SV3SV1SV4NCNOCNONCCCNONCCNCDiluentInlet(A)(B)BallFloatPressurePumpWaste(Vacuum)SV1andSV3areenergized.PressurePumpisactivated.Pressureisappliedtotheapertureto removetheclog.Thesamefunctionoccursifthe FLUSHkeyispressed.ClogremovalOperationSequencesHistogramSOperationSequencesHistogramSV2SV3SV1SV4NCNOCNONCCCNONCCNCDiluentInlet(A)(B)BallFloatOpentotheAirWaste(Vacuum)Whenpowerswitchisturnedon, SV1andSV4areenergized.Diluentisaspiratedthroughthe transducer.Thesamefunctionoccursifthe FILLkeyispressed.Thediluentremovesdiluentandany bloodsampleremaininginthesystem.DiluentFillOperationSequencesHistogramSHaemoglobinHistogramHaemoglobinisthedyeattheRBC.ItisaFe-containingProtein.HaemoglobinisexclusivelyproducedbyErythroblastsinthebonemarrow.HemoglobinisanimportantpartoftheErythrocytes.ThereisadirectrelationbetweenHaematokrit,Erythrocyte-concentrationandHaemoglobin.ThemainfunctionofHaemoglobininthebodyistotransportOxygenandCO2.HaemoglobinHistogramHaemoglobiHaemoglobinHistogramNewborn 14-26g/dlChildren 10-26g/dlWomen 12-16g/dlMen 14-18g/dl

02468101214161820NewbornBabiesChildrenWomenMenPatientLimitsHaemoglobinHistogramNewborn 14HaemoglobinHistogramAspecialreagentisneeded,containingCyanide

forlysingRBCF-Series=Quicklyser

SysmexKCN-MethodHaemoglobinHistogramSysmexKCNHaemoglobinHistogramSLSmeansSodiumlaurylsulfateSLSinsnottoxic(ingredientinalotofsoapstooth-paste)SysmexSulfolyserisnowusedwiththemajorityof

instruments(K-,E-,NE-,SF-,SE-,XE-systems)OnlySysmexhematologysystemshaveastableKCN-freeHaemoglobin-measurementasstandard.SysmexSLS-MethodHaemoglobinHistogramSLSmeansHaemoglobin-MeasurementaabbHeamoglobinmoleculeHydrophobicareaHydrophilicareaSO3-Na+HaemoglobinHaemoglobin-aabbHeamoglobinmoaabbHaemoglobinmoleculeaHydrophobicareaHydrophilicareaSO3-Na+HaemoglobinCyanide-freeSLS-methodaabbHaemoglobinmoleculeaHydroHydrophobicareaofSLSbindtoGlobinConformationchangeOxidation:Fe2+-->Fe3+HydrophilicareaofSLSbindtoFe3+

-->SLS-Hb.Fe

2+Fe

3+Fe

3+SLSHaemoglobinCyanide-freeSLS-methodHydrophobicareaofSLSbindtphotometricmeasurementat540nm(F-Series,K-800,K-1000,K-4500)LampLensFilterSampleCellpackFlowcellPhotosensorHaemoglobinCyanide-freeSLS-methodphotometricmeasurementat540LampHaemoglobinLampLensFilterDiluentFlowCellPhotoSensorFirststep:BLANKMEASUREMANTLampHaemoglobinLampLensFiltLampHaemoglobinLampLensFilterDiluentFlowCellPhotoSensorSampleLampLensFilterSampleFlowCellPhotoSensorSecondstep:MEASUREMENTWITHLYSEDSAMPLELampHaemoglobinLampLensFiltLampHaemoglobinStartFlowCellisrinsedwithDiluent.MeasurementandsavingofBlank.MeasurementofsampledilutionSamplevalue-Blank=HB-valuePrintingofresultEndLampHaemoglobinStartFlowCellHaematocritHaematocritisdeterminedbycomparingthetotalorcumulativevolumeofredbloodcellstothevolumeofwholeblood.DeterminednormallybycentrifugationorbyelectricpulsemeasurementHaematocritisbesidesHaemoglobinanimportantparameterinanemia-diagnosis.HaematocritHaematocritHaematocritisdeteHaematocrit00,10,20,30,40,50,6NewbornChildrenMenBabiesWomenNewborn 0,51-0,65Children 0,35-0,43Women 0,38-0,48Men 0,42-0,52

PatientLimitsHaematocrit00,10,20,30,40,50,6HaematocritDeterminingthevolumeofcorpusclesinthebloodNormallyequivalenttomassoferythrocytes.Bloodisdrawnintospecialcapillaries,whichcontaindriedheparin.Afterclosing,capillariesareplacedintoacentrifuge.Inthecentrifugebloodisdividedintoitscorpuscular(cells)andfluid(plasma)componentsCentrifugationHaematocritDeterminingthevolHaematocrit-MeasurementHaematocritHaematocrit-HaematocritCentrifugationHaematocritCentrifugationHaematocritTotalvolumeVTCentrifugationHaematocritTotalvolumeVTCentrifugationHCentrifugeCentrifugationHaematocritTotalvolumeVTCentrifugeCentrifugationHaematCentrifugeVTVHct(%)=V/VTx100CentrifugationHaematocritTotalvolumeVTCentrifugeVTVHct(%)=V/VTCentrifugationHaematocritCentrifugationReadingisdonewithascaleoutoftriangles.00,10,20,30,40,50,60,70,80,91,0Theauxiliarylinethroughtheborder

betweenerythrocytes-andleukocytes

givestheheamatocrit.CVapp.1%CentrifugationHaematocritCentrCentrifugationHaematocritPulseHeightTransducerCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightTransducerStart-SensorStop-SensordefinedvolumeVTCentrifugationHaematocritPulseCentrifugationHaematocritPulseHeightPhPulseDiagramPh=kxVEryVEry=1/kxPh

PhPulsehightkConstantVEry Volumeof1ErythrocyteCentrifugationHaematocritPulseCentrifugationHaematocritCumulativePulseHct(%)=V/VTx100V =?VEry

PhPulsehightkConstantVEry Volumeof1ErythrocyteVTVCentrifugationHaematocritCumulTransducerPh=kxVEryVEry=Phx1/kPh PulseHeightk KonstantVEry VolumeEry

PhHct(%)=V/VTx100V=?VEry=1/kx?PhHct(%)=VT1/kx?Phx100VVTCumulativePulseHeightDetectionK-SeriesHaematocritTransducerPh=kxVEryPhHct(ErythrocyteIndicesCalculated(Parameters)ErythrocyteIndicesErythrocyte-IndicesMeanCorpuscularVolume(MCV)Hct(%)RBC(x106/μl)MCV(fl)=MeanCorpuscularHaemoglobin(MCH)Hb(g/dl)RBC(x106/μl)MCH(pg)=MeanCellularHaemoglobinConcentration

(MCHC)

Hb(g/dl)Hct(%)MCHC(g/dl)=RBC-IndicesErythrocyte-IndicesMeanCorpusBiologicalVariation-40-20020406080100ofquantitativeparametersofCBCDeviationfromMedianin%WBC103/μl5,0190PLT103/μl14Hgbg/dl4,5RBCx106/μl42HCT%90MCVfl34MCHCg/dl31MCHpgBiologicalVariationBiologicalVariation-40-200204SpecificationsExample:SYSMEXK-4500SpecificationsSpecificationsSpecifications18Parameter:

WBC,RBC,HBG,HCT,PLT

MCV,MCH,MCH

RDW-CV,RDW-SD,MPV,PDW,P-LCR

SCR%(LYMPH%),MCR%(MONO/EO/BASO%),LCR%(NEUT%)

SCC#(LYMPH#),MCC#(MONO/EO/BASO#),LCC#(NEUT#)

3Histograms:

WBC,RBC,PLTSpecifications18Parameter:

WBC,RBC,HBG,HReliableMeasurementPrinciplesfor

highestaccuracyWBC/RBC/PLT DCDetectionMethodHBG SLS-HaemoglobinMethodHCT CumulativePulseHeightDetection WBC-Diff DCDetectionMethodSpecificationsReliableMeasurementPrincipleThroughput80Samples/HourCycleTimewithsingleSamples45Seconds/SampleSpecificationsThroughputSpecificationsSampleVolumeClosedMode(Sampler) 200μlManualOpenMode 100μlMCP-Mode 200μlCapillaryMode >40μSpecificationsSampleVolumeSpecificationsSpecificationsParameters WBC,NEUT%,LYMPH%,EO/MONO/BASO%, NEUT#,LYMPH#,EO/MONO/BASO#,RBC,HKT, HGB,MCV,MCH,MCHC,RDW-SD,RDW-CV,PLT, PDW,MPV,P-LCRReagents Diluent CELLPACK Lyse-Reagent STROMATOLYSER-3WP haemoglobin-Reagent SULFOLYSER CleaningReagent(Detergent) CELLCLEANDiameterofApertures WBC 100μm RBC/HCT/PLT 75μmThroughput 80Samples/Hour (appr.45SecondsCycle-Time/Sample)Principles - DCDetectionMethod(RBC,WBC,PLT) - SLS-Hemoglobin(HGB)

- CumulativePulseHeightDetection - CalculationfromRBCandHCT(MCV) - CalculationfromRBCandHGB(MCH) - CalculationfromHCTandHGB(MCHC)SampleVolume SamplerMode 200μl ClosedMode 200μl ManualOpenMode 100μl

MCP-Mode 200μl CapillaryBloodMode >40μlDataMemory Externaldatamemoryavailable(K-DPS)SpecificationsParameters WBC,-Precision Onefreshnormalwholebloodsamplewasmeasured10times consecutivelyinSamplerMode.Fromthoseresultsthe CoefficientofVariationhasbeencalculated.

WBC CV<3,0% RBC CV<1,5% HGB CV<1,5% HCT CV<1,5% MCV CV<1,5% MCH CV<1,5% MCHC CV<1,5% PLT CV<4,0%Accuracy WBC ±3,0% RBC ±3,0% PLT ±3,0%Linearity- WBC within±0,2x103/μlfor1,0-6,6x103/μl inSamplerMode or3,0%für6,7-99,9x103/μl RBC within±0,03x106/μlfor0,30-0,99x06/μl or3,0%for1,0-99,9x106/μl HGB within±0,1g/dlfor0,1-10,0g/dl or1,0%for10,1-20,0g/dl and2,0%for20,1-25,0g/dl HCT within±1,0%for10,0-33,3% or2,0%for33,4-60,0% PLT within±10X10^3/μlfor10-199x103/μl or5,0%for200-999x103/μl (ifRBC<7,00x106/μl)Blank WBC <0,3x103/μl RBC <0,02x106/μl HBG <0,1g/dl PLT <10x103/μlSpecifications-Precision OnefreshnormalwhDisplayRange WBC 0,00-300,0x103/μl RBC 0,00-20,00x106/μl HGB 0,0-25,0g/dl HCT 0,0-100% PLT 0-2000x103/μlPorts SerialportforHostorK-DPS ParallelportfordataprinterDP-510/490Size MainUnit 580x600x31HeightxWidthxDepth Sampler 580x302x280[mm]PowerSupply 230VAC/DCat50HzBarcode-Reading CODABAR,CODE-11,CODE-39,CODE-128,Interleaved2of5, JAN-8,JAN-13SpecificationsDisplayRange WBC 0,00-300,0SampleFlowSampleFlowSampleFlowSampleFlowStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)CellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)20ml4μlCellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)RBCChannel1:25.00040μl2,0mlCellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)WBCChannel1:2501,0ml0,5ml6μlCellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)HgbPhotometer1:5003μl1,0ml0,5mlCellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixStromatolyser-3WPSulfolyserMixing-Chamber1:500SampleRotorValve(SRV)20ml4μlRBCChannel1:25.00040μl2,0mlWBCChannel1:2501,0ml0,5ml6μlHbPhotometer1:5003μl1,0ml0,5mlCellpackSampleFlowFlowChartStromatolyser-3WPSulfolyserMixMiddlePart(moving)UpperPart(fixed)LowerPart(fixed)PipetteSampleFlowSampleRotorValveSampleRotorValveMiddlePart(moving)UpperPartSampleRotorValveMiddlePart(moving)UpperPart(fixed)LowerPart(fixed)PipetteSampleFlowSampleRotorValveSampleRotorValveMiddlePartMiddlePart(moving)UpperPart(fixed)LowerPart(fixed)PipetteSampleFlowSampleRotorValveSampleRotorValveMiddlePart(moving)UpperPartWholeBloodSampling0,1mlWholeBloodVacuumbyDiaphragm-PumpSampleFlowSampleRotorValveWholeBloodSampling0,1mlWhoSampleSeparation12μl(WBC)4μl(RBC)6μl(Hb)SampleFlowSampleRotorValveSampleSeparation12μl(WBC)4μRBC-DilutionDiluent-Diaphragm-PumpMixing-Chamber4μl(RBC)SampleFlowSampleRotorValveRBC-DilutionDiluent-Diaphragm-Samplingfor2ndRBC-Dilution2ndDilution&Diaphragm-PumpMixing-ChamberSampleFlowSampleRotorValveSamplingfor2ndRBC-Dilution2Separationfor2ndRBC-Dilution40μlSampleFlowSampleRotorValveSeparationfor2ndRBC-Dilutio2ndRBC-DilutionDiluent-Diaphragm-PumpRBC-ChannelSampleFlowSampleRotorValve2ndRBC-DilutionDiluent-RBC-ChWBC-andHGB-SampleDilution12μl(WBC)6μl(HGB)Diluent-Diaphragm-PumpHGB-Diaphragm-PumpWBC-ChannelHGB-FlowCellSampleFlowSampleRotorValveWBC-andHGB-SampleDilution1HistogramsHistogramsInterpretationNO.4 DATE: 9/10/95 15:11MODE: WholeBloodWBC 5,8 x103/μlRBC 4,84 x106/μlHGB 13,7 g/dlHCT 42,0 %MCV 86,8 flMCH 28,3 pgMCHC 32,6 g/dlPLT 257 x103/μlLYMPH% 31,2 %MXD% 6,8 %NEUT% 62,0 %LYMPH# 1,8 x103/μlMXD# 0,4 x103/μlNEUT# 3,6 x103/μl250RBCRDW-SD 40,0 fl40PLTPDW 13,1 flMPV 10,4 flP-LCR 28,1 %WBC300HistogramsHistogramsInterpretNormalSampleNO.4 DATE: 9/10/95 15:11MODE: WholeBloodWBC 5,8 x103/μlRBC 4,84 x106/μlHGB 13,7 g/dlHCT 42,0 %MCV 86,8 flMCH 28,3 pgMCHC 32,6 g/dlPLT 257 x103/μlLYMPH% 31,2 %MXD% 6,8 %NEUT% 62,0 %LYMPH# 1,8 x103/μlMXD# 0,4 x103/μlNEUT# 3,6 x103/μl250RBCRDW-SD 40,0 fl40PLTPDW 13,1 flMPV 10,4 flP-LCR 28,1 %WBC300ParametersforCBCLeukocyte-Histogram

Lymphocytesgivenin%andabsolutevalue

Eo,Mono,Basogivenin%andabsolutevalue

Neutrophilsgivenin%andabsolutevalueErythrocyte-Histogram

ErythrocyteDistributionWidthPlatelet-Histogram

PlateletDistributionWidth

MeanPlateletVolume

Platelet-LargeCellRatioWBCRBCPLTHistogramsInterpretationNormalSampleNO.4 LYMPH% 31RBC-andPLT-Histograms2fl25-30fl250flThetwodistributioncurvesareseparatedfromeachotherbyamovingautodiscriminatorlookingtothePlateau.Plateletshaveasizebetween8and12flandarecountedbetween2and30fl.Erythrocyteshaveasizeof80-100flandarecountedbetween25and250fl.HistogramsRBC-andPLT-Histograms2fl25-RBC-andPLT-HistogramsTheSizeDistributionCurveshouldalwaysstartonthebaselineandfallbetweenthelowerandtheupperdiscriminator.2fl250flBaselineErythrocytesHistogramsRBC-andPLT-HistogramsTheSizPLTLDRBCThecurvedoesnotstartatthebaseline.Mark“RL“,abnormalheightatlowerdiscriminatorPossiblecauses:GiantPlateletsMicro-ErythrocytesPlateletClumpsErythrocyteHistogramFlagsHistogramsErythrocytesPLTLDRBCThecurvedoesnotstaErythrocytesUDRBCThecurvedoesnotendatthebaseline.UDRBCPossiblecauses:ColdAgglutininsErythroblasts/NormoblastsErythrocyteHistogramFlagsHistogramsMark“RU“,abnormalheightattheupperdiscriminator.ErythrocytesUDRBCThecurvedoErythrocytes“MP“,multiplepeaksfoundRBCErythrocyteHistogramFlagsRBCHistogramsErythrocytes“MP“,multipleErythrocytes“DW“,abnormalhistogramdistributionRBCRBCDistributioncurvedoesnotcross20%leveltwiceErythrocyteHistogramFlagsHistogramsErythrocytes“DW“,abnormalhErythrocytes100%ssμRDW-CV(%)=100xs/μ68,26%RDW-CV11-16%100%20%RDW-SD37-46flRedBloodCellDistributionWidthHistogramsErythrocytes100%ssμRDW-CV(%)30fl2DiscriminatorsThrombocyteHistogramHistograms30fl2DiscriminatorsThrombocyPLT,Countofplatelets

HistogramparametersMPV,meanplateletvolume

Referencerange:8-12flP-LCR,ratiooflargeplatelets

Referencerange15-35%PDW,plateletdistributionwidthat20%ofpeakheight

Referencerange:9-14fl

Pct(%)PLT(x103/μl)MPV(fl)=12flLDUDPLTP-LCR100%20%PDWThromboyteHistogramHistogramsPLT,Countofplatelets

Pct(Mark“PL“,abnormalheightatlowerdiscriminatorPLTCurvedoesnotstartfrombaselinePLTPossiblecause:HighblankvalueCellfragmentsThrombocyteHistogramFlagsHistogramsMark“PL“,abnormalheightaCurvedoesnotreachbaselinePossiblecause:ClottedbloodsampleEDTA-inducedPLTclumpingMicro-ErythrocytesGiantPlateletsPlateletaggregatesHistogramsThrombocyteHistogramFlagsMark“PU“,abnormalheightatupperdiscriminatorCurvedoesnotreachbaselinePHistogramsMark“MP“,MultiPeaksfoundPossibleCause:PlatelettransfusionThrombocyteHistogramFlagsHistogramsMark“MP“,MultiHistogramsMark“DW“,DistributionWithPLTThedistributioncannotbedetectedbecausetheHistogramdoesnotcrossthe20%limittwice.Thiscurveinonlyanexamplebutcouldalsoshowanothercourse.ThrombocyteHistogramFlagsHistogramsMark“DW“,DistriHistogramLyseofRBCandpartiallyseofWBCWBCMitochondriaNucleusCellmembraneRibosomeCytoplasmElectrolytesolutionAfterlysisLeukocyte-Histogram

HistogramLyseofRBCandpartHistogrammBeforeaddinglysingreagent0 2 4 6 8 10 12 14 16 18 20 22NeutrophilsBasophilsEosinophilsMonocytesLymphocytesCelldiameterinμm10-159-1411-1612-207-12Leukocyte-Histogram

Afteraddinglysingreagent050100150 200 250 300 LymphocytesMIXMonocytesBasophilsEosinophilsNeutrophils30-8060-12070-13080-140120-250CellsignalinflLymphocytesMonocytesBasophilsEosinophilsNeutrophilsHistogrammBeforeaddinglysinHistogramIMPORTANT:

Thedistributioncurveshouldbewithinthediscriminators.Thecurveshouldstartandendatthebasisline.

TheLDisflexible,butcannotbelowerthan30fl.

TheWBC-channelshowLeukocytesand

Thrombocytes(Erythrocytesarelysed).

ThevolumeoftheThrombocytsisusuallybetween8-12fl,thereforetheLDattheWBC-HistogrammseperatestheLeukocytesfromtheThrombocytes.(Thrombocyteswerenotcounted).Curveislimitedby2discriminators.050100150 200 250 300 UD(fix)T2T1LDLeukocyte-Histogram

HistogramIMPORTANT:CurveisHistogram1.Flag“WL“,CurvedoesnotbeginatthebasislinePossiblecause:Thrombocytes

EDTA-notcompatible

Probecoagulatedhighosmoticresistant(Erythrocytesnotlysed)Normoblasten/ErythroblastencoldagglutinateLeukocytes-HistogramFlagsHistogram1.Flag“WL“,CurveHistogram2.Flag“WU“,Curvedoesnotendatthebaseline. Sampledilution?(highleukocytecount?)

Leukocytes-HistogramFlagsHistogram2.Flag“WU“,CurvHistogram3.Flag“T1”and“T2”T1andT2aretroughdiscriminatorsdefinedbytheplateau.ThisdiscriminatorsseparatestheLeukocytespopulations.050100150 200 250 300 UD(fix)T2T1LDPopulation1=F1Population2=F2

Population3=F3F=FractionThediscriminatorsareflexibleandwillbesetautomaticallyaccordingtothesample.

Inspecialcasesisaseparationfromthetroughdiscriminatorsnotpossible.Leukocytes-HistogramFlagsHistogram3.Flag“T1”and“T23.Flag“T1”and“T2”T1couldnotbedetectedNoplateau/troughwasfound.-->T1flagT1wasdetectedbutnotT2

-->T2flagAttention:

ConfirmtheresultwiththemicroscopeifT1orT2flagwasindicated.TheWBCresultwillbecorrectifnoflagisindicated.AllLeukocytesarecounted.HistogramLeukocytes-HistogramFlags3.Flag“T1”and“T2”T1coul3.Flag“F1”,“F2”and“F3”TheHistogramoftheLeukocytesislimitedfromtheouterdiscriminatorsLDandUD.

F1F2 F3Thereisapotentialofmixingpopulations.F1andF2movetogether.AlsoF2andF3.HistogramLeukocytes-HistogramFlagsAllLeukocytesarecounted;WBCtotaliscorrect.

(Assumption:nootherflags)T1andT2weredetected.Conspicuousis:

Thetroughsareawayfromthebasisline.3.Flag“F1”,“F2”and“F3”ThMaintenanceMaintenanceInterpretationMaintenanceMaintenanceInterprMaintenance1. ExternalCheckIsthereenoughreagentforroutinedailyanalyses?

CELLPACK ca.750Cycles/20l

STROMATOLYSER-3WP ca.1.000Cycles/0,5l

SULFOLYSER ca.1.000Cycles/0.5l

Enoughprinting-paper?

Tubingandwiringconnectedcorrectly?

Sampler*ready?

(*optional)Wastecontainerempty?Maintenance1. ExternalCheckMaintenance2. Switc