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1、DC Detection Method,The Pulse,First Dilution 1:500(WBC)Second Dilution 1:50.000(RBC),Sample BeakerDB-1,Dilutions for Semiautomatic Analyser,DC Detection Method,The Pulse,WBC Dilution+3 Drops Quicklyser for Lysing and HGB,RBC Dilution,Dilutions for Semiautomatic Analyser,DC Detection Method,The Pulse
2、,External Electrode,Aperture,Internal Electrode,DC Detection Method,Transducer,Vacuum,Blood cells,External Electrode,DC Detection Method,Transducer,External Electrode,Aperture,Internal Electrode,DC Detection Method,ExternalElectrode,Aperture,Internal Electrode,U=R x I,The Pulse,DC Detection Method,U
3、=R x I,ExternalElectrode,Aperture,InternalElectrode,The Pulse,DC Detection Method,Time sec,Blood suspension,Transducer,Start-Sensor,Stop-Sensor,Manometer,Counting Time,(defined Volume),forerun,afterrun,Absolute Counting I,Time sec,Blood suspension,Transducer,Start-Sensor,Stop-Sensor,Manometer,Counti
4、ng Time,(defined Volume),Prerun,Afterrun,DC Detection Method,Absolute Counting I,Time sec,Blood suspension,Transducer,Start-Sensor,Stop-Sensor,Manometer,Counting Time,(defined Volume),Prerun,Afterrun,DC Detection Method,Absolute Counting I,Time sec,Blood suspension,Transducer,Start-Sensor,Stop-Senso
5、r,Manometer,Counting Time,(defined Volume),Prerun,Afterrun,DC Detection Method,Absolute Counting I,Time sec,Blood suspension,Transducer,Start-Sensor,Stop-Sensor,Manometer,Counting Time,(defined Volume),Prerun,Afterrrun,DC Detection Method,Absolute Counting I,DC Detection Method,Absolute Counting,DC
6、Detection Method,Absolute Counting II,Not to be changed by User:Sample VolumeDilution RatioCounted VolumeAperture DiameterAutomatically checked by the instrument:Counting TimeCondition of aperture during measurement(Noise-Control)Advantages:No calibration of countersAutomatic check of aperture,DC De
7、tection Method,Relative Counting,DC Detection Method,Measurement of particlesin a defined volumeCalculation of cell-concentration with known Sample volume and defined Sample dilutionno calibration of counting values,Measurement of particles per time periodThe count is received indirectly through a r
8、eference-sample.It is taken for granted that the sample and the reference-sample have the same characteristics and give the same result.Calibration of counting values in necessary,Absolute Counting,Relative Counting,Comparison of Absolute and Relative Counting,Pulse Height,Volume(Time),DC Detection
9、Method,1,2,3,4,5,6,7,8,9,10,11,12,13,14,Cumulative Size Distribution Curve,Pulse Picture,10,20,30,1,2,3,4,5,6,7,8,9,10,11,12,13,14,4,1,0,0,0,1,2,3,4,5,4,3,2,1,DC Detection Method,Histogram,1,2,3,4,5,6,7,8,9,10,11,12,13,14,Histogram,4,1,0,0,0,1,2,3,4,5,4,3,2,1,Cumulative Size Distribution Curve,10,20
10、,30,1,2,3,4,5,6,7,8,9,10,11,12,13,14,4,1,0,0,0,1,2,3,4,5,4,3,2,1,DC Detection Method,Histogram,Erythrocyte Histogram,25-75 fl,200-250 fl,DC Detection Method,Histogram,Platelet Histogram,2-6 fl,12-30 fl,fixed at12 fl,DC Detection Method,Histogram,DC Detection Method,Histogram,Operation Sequences,Hist
11、ogram,All solenoid valves are de-energized.C(Common)and NO(Normally Open)solenoid valve ports are connected.Ball Float is in the down position.,“READY”-Status,Operation Sequences,Histogram,COUNT KEY on,SV1 and SV 4 are energized.C and NC solenoid valve ports are connected.Diluent is aspirated throug
12、h SV4,SV2,Transducer,SV1 and SV3.The Ball Float in the manometer ispushed upward.The inner portion of transducer and manometer is automatically rinsed to minimize carry-over effect.,Operation Sequences,Histogram,Counting,SV1 and SV4 are de-energized and SV2 is energized.The diluted sample is aspirat
13、ed throughthe aperture and the Ball Float moves downward.When the Ball Float reaches level A,the counter starts counting.When the Ball Float reaches level B,the counting stops and SV2 is de-energized.The volume of aspirated sample is the same as the manometer-volume between points A and B.,Operation
14、 Sequences,Histogram,SV1 and SV3 are energized.Pressure Pump is activated.Pressure is applied to the aperture to remove the clog.The same function occurs if the FLUSH key is pressed.,Clog removal,Operation Sequences,Histogram,When power switch is turned on,SV1 and SV4 are energized.Diluent is aspira
15、ted through the transducer.The same function occurs if the FILL key is pressed.The diluent removes diluent and anyblood sample remaining in the system.,Diluent Fill,Haemoglobin,Histogram,Haemoglobin is the dye at the RBC.It is a Fe-containing Protein.Haemoglobin is exclusively produced by Erythrobla
16、sts in the bone marrow.Hemoglobin is an important part of the Erythrocytes.There is a direct relation between Haematokrit,Erythrocyte-concentration and Haemoglobin.The main function of Haemoglobin in the body is to transport Oxygen and CO2.,Haemoglobin,Histogram,Patient Limits,Haemoglobin,Histogram,
17、A special reagent is needed,containing Cyanidefor lysing RBCF-Series=Quicklyser,Sysmex KCN-Method,Haemoglobin,Histogram,SLS means SodiumlaurylsulfateSLS ins not toxic(ingredient in a lot of soaps tooth-paste)Sysmex Sulfolyser is now used with the majority of instruments(K-,E-,NE-,SF-,SE-,XE-systems)
18、Only Sysmex hematology systems have a stable KCN-free Haemoglobin-measurement as standard.,Sysmex SLS-Method,Haemoglobin-Measurement,Heamoglobin molecule,Hydrophobic area,Hydrophilic area,SO,3,-,Na,+,Haemoglobin,a,a,b,b,Haemoglobin molecule,a,Hydrophobic area,Hydrophilic area,SO,3,-,Na,+,Haemoglobin
19、,Cyanide-free SLS-method,Hydrophobic area of SLS bind to GlobinConformation changeOxidation:Fe2+-Fe3+Hydrophilic area of SLS bind to Fe3+-SLS-Hb.,Fe 2+,Fe 3+,Fe 3+,SLS,Haemoglobin,Cyanide-free SLS-method,photometric measurement at 540 nm(F-Series,K-800,K-1000,K-4500),Lamp,Lens Filter,Sample,Cellpack
20、,Flowcell,Photosensor,Haemoglobin,Cyanide-free SLS-method,Lamp,Haemoglobin,First step:BLANK MEASUREMANT,Lamp,Haemoglobin,Second step:MEASUREMENT WITH LYSED SAMPLE,Lamp,Haemoglobin,Start,Flow Cell is rinsed with Diluent.,Measurement and saving of Blank.,Measurement of sample dilution,Sample value-Bla
21、nk=HB-value,Printing of result,End,Haematocrit,Haematocrit is determined by comparing the total or cumulative volume of red blood cells to the volume of whole blood.Determined normally by centrifugation or by electric pulse measurementHaematocrit is besides Haemoglobin an important parameter in anem
22、ia-diagnosis.,Haematocrit,Haematocrit,Newborn0,51-0,65Children0,35-0,43 Women 0,38-0,48 Men 0,42-0,52,Patient Limits,Haematocrit,Determining the volume of corpuscles in the bloodNormally equivalent to mass of erythrocytes.Blood is drawn into special capillaries,which contain dried heparin.After clos
23、ing,capillaries are placed into a centrifuge.In the centrifuge blood is divided into its corpuscular(cells)and fluid(plasma)components,Centrifugation,Haematocrit-Measurement,Haematocrit,Centrifugation,Haematocrit,Total volume VT,Centrifugation,Haematocrit,Centrifuge,Centrifugation,Haematocrit,Total
24、volume VT,Centrifuge,VT,V,Hct(%)=V/VT x100,Centrifugation,Haematocrit,Total volume VT,Centrifugation,Haematocrit,Centrifugation,Reading is done with a scale out of triangles.,The auxiliary line through the borderbetween erythrocytes-and leukocytesgives the heamatocrit.CV app.1%,Centrifugation,Haemat
25、ocrit,Pulse Height,Transducer,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,define
26、d volume VT,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centrifugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centri
27、fugation,Haematocrit,Pulse Height,Transducer,Start-Sensor,Stop-Sensor,defined volume VT,Centrifugation,Haematocrit,Pulse Height,Ph,Pulse Diagram,Ph=k x VEry VEry=1/k x Ph Ph Pulse hightk ConstantVEry Volume of 1 Erythrocyte,Centrifugation,Haematocrit,Cumulative Pulse,Hct(%)=V/VT x 100V=VEry,Ph Pulse
28、 hightk ConstantVEry Volume of 1 Erythrocyte,VT,V,Cumulative Pulse Height Detection,K-Series,Haematocrit,Erythrocyte IndicesCalculated(Parameters),Erythrocyte-Indices,RBC-Indices,Biological Variation,-40,-20,0,20,40,60,80,100,of quantitative parameters of CBC,Deviation fromMedian in%,Biological Vari
29、ation,Specifications Example:SYSMEX K-4500,Specifications,18 Parameter:WBC,RBC,HBG,HCT,PLTMCV,MCH,MCHRDW-CV,RDW-SD,MPV,PDW,P-LCRSCR%(LYMPH%),MCR%(MONO/EO/BASO%),LCR%(NEUT%)SCC#(LYMPH#),MCC#(MONO/EO/BASO#),LCC#(NEUT#)3 Histograms:WBC,RBC,PLT,Specifications,Reliable Measurement Principles forhighest a
30、ccuracy,WBC/RBC/PLTDC Detection MethodHBGSLS-Haemoglobin MethodHCTCumulative Pulse Height DetectionWBC-DiffDC Detection Method,Specifications,Throughput80 Samples/HourCycle Time with single Samples45 Seconds/Sample,Specifications,Sample VolumeClosed Mode(Sampler)200 lManual Open Mode100 lMCP-Mode200
31、 lCapillary Mode 40,Specifications,Specifications,ParametersWBC,NEUT%,LYMPH%,EO/MONO/BASO%,NEUT#,LYMPH#,EO/MONO/BASO#,RBC,HKT,HGB,MCV,MCH,MCHC,RDW-SD,RDW-CV,PLT,PDW,MPV,P-LCRReagentsDiluentCELLPACKLyse-ReagentSTROMATOLYSER-3WPhaemoglobin-Reagent SULFOLYSERCleaning Reagent(Detergent)CELLCLEANDiameter
32、 of AperturesWBC100 mRBC/HCT/PLT75 mThroughput80 Samples/Hour(appr.45 Seconds Cycle-Time/Sample)Principles-DC Detection Method(RBC,WBC,PLT)-SLS-Hemoglobin(HGB)-Cumulative Pulse Height Detection-Calculation from RBC and HCT(MCV)-Calculation from RBC and HGB(MCH)-Calculation from HCT and HGB(MCHC)Samp
33、le VolumeSampler Mode200 lClosed Mode200 lManual Open Mode100 lMCP-Mode200 lCapillary Blood Mode 40 lData MemoryExternal data memory available(K-DPS),-PrecisionOne fresh normal whole blood sample was measured 10 times consecutively in Sampler Mode.From those results the Coefficient of Variation has
34、been calculated.WBCCV 3,0%RBCCV 1,5%HGBCV 1,5%HCTCV 1,5%MCVCV 1,5%MCHCV 1,5%MCHCCV 1,5%PLT CV 4,0%AccuracyWBC 3,0%RBC 3,0%PLT 3,0%Linearity-WBCwithin 0,2 x 103/l for 1,0-6,6 x 103/l in Sampler Modeor 3,0%fr 6,7-99,9 x 103/lRBCwithin 0,03 x 106/l for 0,30-0,99 x 06/lor 3,0%for 1,0-99,9 x 106/lHGBwith
35、in 0,1 g/dl for 0,1-10,0 g/dlor 1,0%for 10,1-20,0 g/dland 2,0%for 20,1-25,0 g/dlHCTwithin 1,0%for 10,0-33,3%or 2,0%for 33,4-60,0%PLTwithin 10 X 103/l for 10-199 x 103/lor 5,0%for 200-999 x 103/l(if RBC 7,00 x 106/l)BlankWBC 0,3 x 103/lRBC 0,02 x 106/lHBG 0,1 g/dlPLT 10 x 103/l,Specifications,Display
36、 Range WBC0,00-300,0 x103/lRBC0,00-20,00 x 106/lHGB0,0-25,0 g/dlHCT0,0-100%PLT0-2000 x 103/lPortsSerial port for Host or K-DPSParallel port for data printer DP-510/490Size Main Unit580 x 600 x 31Height x Width x DepthSampler580 x 302 x 280mmPower Supply230 V AC/DC at 50 HzBarcode-ReadingCODABAR,CODE
37、-11,CODE-39,CODE-128,Interleaved 2 of 5,JAN-8,JAN-13,Specifications,Sample Flow,Sample Flow,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),Cellpack,Sample Flow,Flow Chart,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),Cellpack,Sample Flow,Flow Char
38、t,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),Cellpack,Sample Flow,Flow Chart,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),Cellpack,Sample Flow,Flow Chart,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),Cellpack,Sample
39、 Flow,Flow Chart,Stromatolyser-3WP,Sulfolyser,Mixing-Chamber1:500,Sample Rotor Valve(SRV),WBC Channel1:250,1,0 ml,0,5 ml,6 l,Hb Photometer1:500,3 l,1,0 ml,0,5 ml,Cellpack,Sample Flow,Flow Chart,Lower Part(fixed),Pipette,Sample Flow,Sample Rotor Valve,Sample Rotor Valve,Sample Rotor Valve,Middle Part
40、(moving),Upper Part(fixed),Lower Part(fixed),Pipette,Sample Flow,Sample Rotor Valve,Middle Part(moving),Upper Part(fixed),Lower Part(fixed),Pipette,Sample Flow,Sample Rotor Valve,Sample Rotor Valve,Whole Blood Sampling,0,1 ml Whole Blood,Vacuum by Diaphragm-Pump,Sample Flow,Sample Rotor Valve,Sample
41、 Separation,12l(WBC),4 l(RBC),6 l(Hb),Sample Flow,Sample Rotor Valve,RBC-Dilution,Diluent-Diaphragm-Pump,Mixing-Chamber,4 l(RBC),Sample Flow,Sample Rotor Valve,Sampling for 2nd RBC-Dilution,2nd Dilution&Diaphragm-Pump,Mixing-Chamber,Sample Flow,Sample Rotor Valve,Separation for 2nd RBC-Dilution,40 l
42、,Sample Flow,Sample Rotor Valve,2nd RBC-Dilution,Diluent-Diaphragm-Pump,RBC-Channel,Sample Flow,Sample Rotor Valve,WBC-and HGB-Sample Dilution,12l(WBC),6 l(HGB),Diluent-Diaphragm-Pump,HGB-Diaphragm-Pump,WBC-Channel,HGB-Flow Cell,Sample Flow,Sample Rotor Valve,Histograms,Histograms,Interpretation,Nor
43、mal Sample,NO.4DATE:9/10/9515:11MODE:Whole BloodWBC5,8x 103/lRBC4,84 x106/lHGB13,7 g/dlHCT42,0%MCV86,8flMCH28,3pgMCHC32,6g/dlPLT257x103/l,LYMPH%31,2%MXD%6,8%NEUT%62,0%LYMPH#1,8x103/lMXD#0,4x103/lNEUT#3,6x103/l,250,RBC,RDW-SD40,0fl,40,PLT,PDW 13,1flMPV 10,4flP-LCR 28,1%,WBC,300,Parameters for CBCLeuk
44、ocyte-HistogramLymphocytes given in%and absolute valueEo,Mono,Baso given in%and absolute valueNeutrophils given in%and absolute valueErythrocyte-HistogramErythrocyte Distribution WidthPlatelet-HistogramPlatelet Distribution WidthMean Platelet VolumePlatelet-Large Cell Ratio,WBC,RBC,PLT,Histograms,In
45、terpretation,RBC-and PLT-Histograms,The two distribution curves are separated from each other by a moving auto discriminator looking to the Plateau.Platelets have a size between 8 and 12 fl and are counted between 2 and 30 fl.Erythrocytes have a size of 80-100 fl and are counted between 25 and 250 f
46、l.,Histograms,RBC-and PLT-Histograms,The Size Distribution Curve should always start on the base line and fall between the lower and the upper discriminator.,2 fl,250 fl,Base line,Erythrocytes,Histograms,PLT,LD,RBC,The curve does not start at the base line.,Mark“RL“,abnormal height at lower discrimi
47、nator,Possible causes:Giant PlateletsMicro-ErythrocytesPlatelet Clumps,Erythrocyte Histogram Flags,Histograms,Erythrocytes,Erythrocytes,UD,RBC,The curve does not end at the base line.,UD,RBC,Possible causes:Cold AgglutininsErythroblasts/Normoblasts,Erythrocyte Histogram Flags,Histograms,Mark“RU“,abn
48、ormal height at the upper discriminator.,Erythrocytes,“MP“,multiple peaks found,RBC,Erythrocyte Histogram Flags,RBC,Histograms,Erythrocytes,“DW“,abnormal histogram distribution,RBC,Distribution curve does not cross 20%level twice,Erythrocyte Histogram Flags,Histograms,Erythrocytes,RDW-CV(%)=100 x s/
49、,68,26%,RDW-CV11-16%,Red Blood Cell Distribution Width,Histograms,30 fl,2 Discriminators,Thrombocyte Histogram,Histograms,PLT,Count of platelets Histogram parametersMPV,mean platelet volumeReference range:8-12 flP-LCR,ratio of large plateletsReference range 15-35%PDW,platelet distribution width at 2
50、0%of peak height Reference range:9-14 fl,MPV(fl)=,Thromboyte Histogram,Histograms,Mark“PL“,abnormal height at lower discriminator,PLT,Curve does not start from baseline,Possible cause:High blank valueCell fragments,Thrombocyte Histogram Flags,Histograms,Curve does not reach baseline,Possible cause:C
