1. Course title: Hematology (1) Course code : MLHE-201 Supervisor :Prof.Dr Magda Sultan Date : 7-11-2013 Outcome : The student will be able to use the automated cell counter He will know its operating mechanisms He will know the reagents,the alarms and the interfering substances

3. Types: * 3 part diff. (8 or 18 param.) Light transmition Electrical impedance * 5 part diff (23 or 26 param) Laser based cytometer Light scattering

4. 3 part diff. cell counter Counting principle Cell in the counting chamber No particles in the counting chamber + - + -- + + - -

5. Counting principle By counting & sizing cells by detecting and measuring changes in electrical impedance when a cell (or other particle) in a conductive liquid passes through a small aperture. Cells behave as insulators so the resistance change produces a pulse between electrodes as cells pass through the aperture. The amplitude of the pulse is proportional to the cell volume

6. Different Impedance

12. Normal Histogram Three parts differential white blood cells: 40 to 125µ3 : lymphocytes 125 to 160µ3 : monocyte 140 to 400µ3 : granulocytes

15. Reagents needed for cell counter Isotone solu. (dilution and basic impedance). Detergant solu. (prevent carry over and cleaning Hb cuvette). Lyse solu. (lyse RBC for WBCs & palt count). Clean solu. (end day clean). Control blood (quality assurance).

16. Biological and mechanical alarms Biological alarms Assimmetry on one curve of the 3 population of cells (Blood sample abn.) Mechanical alarm Disturbance in the machine No blanking No solution Apperture obstruction Electricity, tubes, pump …

17. Known interfering substance RBCs  High WBCs esp if RBCs is low   RBCs  Agglutinated RBCs   RBCs Hb Turbidity of the blood sample   Hb  Elevated WBCs  Elevated lipids  Fetal bloods Known interfering substance RBCs  High WBCs esp if RBCs is low   RBCs  Agglutinated RBCs   RBCs Hb Turbidity of the blood sample   Hb  Elevated WBCs  Elevated lipids  Fetal bloods

18. MCV  Red cell agglutination   number of large platelets HT  Red cell agglutination RDW  Agglutination of RBCs  Nutritional deficiency  Blood transfusion MCV  Red cell agglutination   number of large platelets HT  Red cell agglutination RDW  Agglutination of RBCs  Nutritional deficiency  Blood transfusion

19. WBCs interferring subs.  Normoblasts   WBCs  Unlysed RBCs   WBCs  MM   WBCs (ppt protein)  Hemolysis   WBCs (red cell stroma)  Leukemia   WBCs (  cell fragility)  In CLL small lymph not counted  Cryoglobulin   all parameters of blood WBCs interferring subs.  Normoblasts   WBCs  Unlysed RBCs   WBCs  MM   WBCs (ppt protein)  Hemolysis   WBCs (red cell stroma)  Leukemia   WBCs (  cell fragility)  In CLL small lymph not counted  Cryoglobulin   all parameters of blood

20. Lymphocytes  Nucleated RBCs   lymph  Parasites   lymph  Resistent RBCs   lymph Monocytes   in large lymphocytes, atypical lymph, blasts and basophils Granulocytes   in eosinophilia, blasts, promyelo, myelo, metameyl and plasma cell Lymphocytes  Nucleated RBCs   lymph  Parasites   lymph  Resistent RBCs   lymph Monocytes   in large lymphocytes, atypical lymph, blasts and basophils Granulocytes   in eosinophilia, blasts, promyelo, myelo, metameyl and plasma cell

21. Platelets  RBCs fragments   plat (microcytes)  WBCs fragments   plat (microcytes)  Chemotherapy   plat (  plat. fragments)  Hemolysis   Plat (red cell strom)  ACD blood   plat (plat. Aggregation)  RBCs inclusion   plat. (Malaria, H.j bodies)  Plat. agglutination   plat Platelets  RBCs fragments   plat (microcytes)  WBCs fragments   plat (microcytes)  Chemotherapy   plat (  plat. fragments)  Hemolysis   Plat (red cell strom)  ACD blood   plat (plat. Aggregation)  RBCs inclusion   plat. (Malaria, H.j bodies)  Plat. agglutination   plat

22. Measured and calculated results Measured results Hb g/dl RBCs / cmm MCV fL WBCs / cmm Plat count / cmm MPV fL Measured results Hb g/dl RBCs / cmm MCV fL WBCs / cmm Plat count / cmm MPV fL Calculated results Ht %: MCV x RBCs Pt %: MPV x plat C MCH: Hb ÷ RBCs MCHC: Hb ÷ Ht RDW %: CV of RBCs Histo PDW %: CV of Pl. Histo Diff WBC 3 relative count 3 abs. count Calculated results Ht %: MCV x RBCs Pt %: MPV x plat C MCH: Hb ÷ RBCs MCHC: Hb ÷ Ht RDW %: CV of RBCs Histo PDW %: CV of Pl. Histo Diff WBC 3 relative count 3 abs. count

23. Normal Histogram

26. Assignment TitleStudent Name Reagents of cell counter.علاء محمد محمود عطيه علي السيد علي بدوي علي عصام علي لبحيصي علي محمد سليم محمد عماد رمسيس سمعان سليمان عمر محمد حسن ابراهيم عمر محمد هاني محمد عمرو عادل عوض عبد الونيس عمرو عبد المنعم احمد عمرو محمد صلاح الدين Types of automated cell counterعمرو محمد محمد عبد الحميد يونس غالي عبد العزيز صالح فاتن لطفي عبد الحميد فطمه صبحي محمود عبد الرحيم فاطمه محمد عبد السلام كريم ابراهيم السعيد كريم احمد خليفه عبد المطلب كريم خالد عبد السلام كريم محمد احمد هاشم ماريان ميلاد قدوس

27. Reference book: Dacie Training Questions What are the reagents of automated cell counter ? Mention the white cell interferring substances