1. Speaker :-Dr Saikat Mandal Moderator :-DrSupriyo Roy Choudhury

2. World Of Automation 3. History Antonie van Leeuwenhoek Invented Microscope The world of cells was colorless until Paul Ehrlich stained blood cells Wallace Coulter :First automated analyzer for counting and sizing cells and presented it in 1956 4. Automation In Hematology Cell counts(Automated hematology analyzers) Diagnosis of hemoglobinopathies(HPLC) Immunophenotyping(Flow cytometry) Coagulation(Coagulometers) 5. Automated Hematology Analyzers Hemogram-Backbone of any lab evaluation As a routine investigation Including anaemia, polycythemia, infection, inflammation, allergy, drug toxicity, malignancy, bleeding tendency etc Aim-to study RBC, WBC series and platelets 6. Advantages Speed with efficient handling of large number of samples Accuracy and precision in quantitative blood tests Ability to perform multiple tests on a single platform Significant reduction of labor requirements Invaluable for accurate determination of red cell indices 7. Disadvantages Flagging of a laboratory test result demands labour intensive manual examination of a blood smear Comments on red cell morphology cannot be generated Platelet Clumps are counted as single. so low count. Erroneously increased or decreased results due to interfering factors Expensive with high running costs 8. Types of counters Semi automated : Some steps carried out manually like dilution of blood Measures only a few parameters Fully automated: Require only anticoagulated blood samples. Measures multiple parameters 9. Components of a cell counter 3 Basic components Hydraulics: Includes aspirating unit, dispencers,diluters,mixing chambers, aperture baths & hemoglobinometer Pneumatics : Vacuums & pressure for operating valves Electronics : Analyzer &computing circuit 10. Principles of Working Electrical Impedance Optical Light Scatter Fluorescence Light absorption Electrical conductivity ?!!@#@# 11. Electrical Impedance First introduced by Wallace Coulter Blood cells are poor conductor of electricity 2 chambers filled with a conductive buffered electrolyte solution Separated by a small aperture DC current between two electrodes 12. Electrical Impedance Diluant displacement causes potential difference Voltage pulse displayed on an osciloscope No. of impulse = No. of cells Height = vol. of cells Freq dist curve & size dist histograms Requisite High dilution 13. Optical Light Scatter Each cell flows in a single line through a flow cell A laser device focussed On striking on cells scattering in different directions Sensor capture & multiplies Forward angle light scatter (FALS)-Cell Size Side scatter(SS)-Granularity 14. Other Methods Peroxide based counters: MPO is used to count neutrophils.Lymphocytes not stained Fluroscence based: Retic and platelet count.Immature pltlets detected best Immunological based: Accurate platelet count using CD41/CD61 antibodies 15. INTERPRETATION 16. 3 Part Analyzer 17. 5 Part Analyzer 18. IP Messages Interpretive messages Assist the laboratory in screening for abnormal samples that may need verification Seen at the bottom end of hemogram 19. Indicators that may appear after the data @ : Data is outside the linearity limit * : Data is doubtful + or :Data is outside the reference limits. ---- : Data doesnt appear due to analysis error or abnormal sample ++++ : Data exceeds display limit. 20. Discrimination Thresholds WBC Discriminator WBC LOWER discriminator-the optimum position in 30 - 60 fL .WBC is calculated from the particle counts more than this LOWER discriminator. RBC Discriminator RBC LOWER discriminator- optimum position in 25 -75 fL and UPPER discriminator, 200 - 250 fL,. RBC is calculated from the particle counts between this LOWER discriminator and UPPER discriminator. PLATELET Discriminator PLT LOWER discriminator, the optimum position in 2 6 fL and and UPPER discriminator- 12 - 30 fL, 21. Histograms These are the graphical presentation of numerical datas of different cell populations in a cell counter.On the X-axis is the cell size and on the Y- axis is the number of cells. Used to determine: The average size Distribution of size Detect subpopulation 22. Flagging System Whenever any significant abnormalities of any cell present , signalled by certain asteriks on the report. Every instrument has its own flagging system. 23. Parameters Measured Directly Measured Derived From Histograms Calculated 1.RBC Count 2.WBC Count 3.Platlet count 4.Hemoglobin 5.Reticulocyte Count 1.MCV 2.RDW 3.DLC 4.PDW 1.Hematorit ( MCV/RBC Count) 2.MCH (Hb/RBC Count) 3.MCHC (Hb/Hct) 24. RBC Histogram Gaussian(Bell Shaped)curve Peak ideally within 80-100 fl 2 flexible discriminator LD (25-75fl) UD(200-250fl) 25. RU-Flag Normoblasts Cold agglutinins ALL- L1 26. RL- Flag Large platelets Fragmented RBCs Platelet aggregation 27. MP-Flag Blood transfusion Dimorphic anaemia Treated IDA 28. Red Cell Distribution Width RDW-SD- 20% height on y axis. Normal-35-45 fl RDW-CV- SD/MCV X 100 Normal- 11.5-14.5 % 29. WBC HISTOGRAM LD(30-60fl) Flexible UD at 300 fl 2 Troughs T1(78-114fl) T2(