blood practical CBC

blood practical CBC

1. BLOOD PRACTICAL CBC

2. Objectives (CBC) At the end of this lesson the students should be able to: Recognize the method used to measure the different hematological values, and compare it with the normal values. Do the calculation of indices, their normal values and their importance in diagnosis of different types of anemia.

3. Aims of the Practical Counting Red blood cells. Counting White blood cells. Determination of hemoglobin concentration. Determination of packed cell volume (PCV) hematocrit. Calculation of red blood cell indices.

4. Complete Blood Count (CBC) It is a test panel requested by a doctor or other medical professional that gives information about the cells in a patient's blood.

5. Complete Blood Count ( cbc)

6. RBC and WBC count

7. Hemocytometer It includes: Neubauer’s slide Cover slip Thoma pipet RBC pipette WBC pipette

8. IMPROVED NEUBAUER’S SLIDE ** It is the name given to a thick glass slide. In the centre of the slide, there is an H- shaped groove . On the two sides of the central horizontal bar, there are scales for counting the blood cells. ** The depth of the scales is 1/10mm or 0.1mm.

9. The numbers written on the chamber mean that the space between the chamber and the cover slip is 0.100 mm and that the smallest square on the grid has an area of 0.0025 mm2. Clean **The numbers written on the chamber mean that the space between the chamber and the cover slip is 0.100 mm and that the smallest square on the grid has an area of 0.0025 mm2.

10. ** Each scale is 3mm wide and 3mm long. ** The whole scale is divided into 9 big squares. ** Each square is 1mm long and 1mm wide.

11. Dr Sitelbanat 2011

12. Differences Between RBC AND WBC Pipette. RBC pipette WBC pipette 1) It has a red bead It has a white bead 2) It has graduations upto mark 101 It has graduations upto mark 11 3) Size of bulb is larger Size of bulb is smaller

13. Thoma Pipet RBC Aspirate to 0.5 or 1.0 Dilute to 101 DF = 200 or 100 WBC Aspirate to 0.5 or 1.0 Dilute to 11 DF = 20 or 10 Discard volume of one (1) in the stem Thoma Pipet

14. RBC PIPETTE WBC PIPETTE

15. Coulter Counter

16. Coulter Counter Principle It count and measure the size of the cells by detecting and measuring electrical resistance when a liquid pass through aperture . While passing the aperture, the cells impedes the current and causes a measurable pulse. Number of pulses --> number of particles. Height of pulses --> volume of particles.

17. Materials and methods Coulter analyzer Diluent reagents Lytic reagent Calibrator kit EDTA anticoagulant blood

18. Diluent –Reagent Is an isotonic electrolyte solution that: - Dilute the whole blood sample - Stabilize cell membrane for accurate counting and size - Conduct aperture current - Rinse instrument components between analysis - Prevent duplicate cell counts Lytic reagent Lysis RBCs for WBCs count and hemoglobin measuremen t

19. Calibrator Kit Is an alternative to the whole blood reference method for calibration

20. RBC, WBC cell count & HB 5ml of venous blood will be drawn in EDTA anticoagulant tube. Diluted by the reagent I and used to count RBC. Lysing RBC using reagent II and used for counting WBC and Hb.

21. Male Female Average RBC 4.5-6.5 x10 6 /  l 3.8-5.8 x10 6 /  l 4.7–6.5 x10 6 /  l WBC 4 – 11 x10 3 /  l 4 – 11 x10 3 /  l 4 – 11 x10 3 /  l HB 13-18 g/dl 11.5-16.5 g/dl 13 –18 g/dl Platelet 150-400x10 3 /  l 150-400x10 3 /  l 150-400x10 3 /  l Normal values

22. Clinical terms  RBC = aneamia  RBC = polycythemia  WBC = leucopenia  WBC = leucocytosis  Platelets = thrombocytopenia  Platelets = thrombocytosis

23. Clinical applications Low numbers of RBCs may indicate: Blood loss: Anemia (various types). Hemorrhage. Bone marrow failure (for example, from radiation, toxin, fibrosis, tumor). Erythropoietin deficiency (secondary to renal disease). Hemolysis (RBC destruction).

24. High numbers of RBCs may indicate: Low oxygen tension in the blood Congenital heart disease Pulmonary fibrosis Polycythemia vera . Dehydration (such as from severe diarrhea). Renal (kidney) disease with high erythropoietin production.

25. High numbers of WBCs (Leucocytosis) may indicate: Infectious diseases. Inflammatory disease (such as rheumatoid arthritis or allergy). Leukemia. Severe emotional or physical stress. Tissue damage (burns).

26. Low numbers of WBCs (leukopenia) may indicate: Bone marrow failure (for example, due to infection, tumor or fibrosis). Presence of cytotoxic substance. Autoimmune/collagen-vascular diseases (such as lupus erythematosus). Disease of the liver or spleen. Radiation exposure.

27. Dr Sitelbanat 2011

28. Dr Sitelbanat 2011

29. Platelet count Thrombocytosis : - Chronic myeloid leukemia. Thrombocytopenia : - Aplastic anemia. - Chemotherapy.

30. Packed Cell Volume (PCV) Hematocrit The ratio of packed blood cells volume to plasma.

31. Procedure Capillary blood obtained from pricking finger tip after cleaning it with alcohol Fill a heparinized capillary tube, then seal one end by plasticine. Centrifuge for 5 minutes to packed the cells at one end of the tube leaving a clear plasma on top. Use the hematocrit reader to find the packed cell volume.

32. Hematocrit Reader

33. Male Female Average PCV % 40-54 35-47 35-54 Normal value PCV

34. Clinical applications High hematocrit may indicate: Dehydration Burns Diarrhea Polycythemia Vera. Low oxygen tension (smoking, congenital heart disease, living at high altitudes).

35. Low hematocrit may indicate: Anemia (various types). Blood loss (hemorrhage). Bone marrow failure (for example, due to radiation, toxin, fibrosis, tumor). Hemolysis (RBC destruction) related to transfusion reaction. Leukemia.

36. The calculation of Red Blood Indices Mean cell volume (MCV) The average volume of red blood cell MCV = PCV x 10 = 88 + 10  m 3 ( fl ) RBC count if the MCV above normal then RBCs is called macrocytes RBCs. ● if the MCV less than normal , RBCs is called microcytic RBCs.

37. MCH less than lower limit of normal: hypochromic RBCs ● MCH greater than upper limit of normal: hyperchromic RBCs Dr Sitelbanat 2011 Mean cell hemoglobin (MCH) The average weight of Hb in red cells MCH = Hb x 10 = 29.5 + 2.5 pg RBC count.

38. Mean cell Hb concentration (MCHC) Concentration of Hb per 100 ml of RBC MCHC = Hb x 100 = 33 + 3 g/dl PCV

39. Average MCV 78-98 m3 MCH 27-32 pg MCHC 30-35 g/dl Normal values

40. Thank you