TOPIC 8 BLOODdshjasjfnlaihfjnsfkja CELL COUNT.pptx

blood cell count

1. Topic 8: Blood Cell Count BT1103 HEMATOLOGY DIPLOMA IN MLT BAINUN DALI ICAN COLLEGE

2. Learning outcome Identify the component and function of Haemocytometer set and Neubauer Counting Chamber Able to Perform blood dilution, incubation and cell counting techniques. Able to understand calculation for total red cell, total white cell and platelet counts State the precautions in cell counting

3. DEFINITION Hemo : blood, cyto : cell, meter: measurement/counter. Thus, it is an instrument used to count the blood cells.

4. Principle of cell counting The blood is diluted with appropriate known volume of diluting fluid and then counting is done by using haemocytometer  

5. 1. HEMOCYTOMETER ( Neubauer chamber )

6. 2. BLOOD DILUTING PIPETTES RBC WBC

7. 3. MICROSCOPE

8. PROCEDURE FOR TOTAL RED CELL & PLATELET COUNT FILLING THE PIPETTE Mix blood thoroughly Use the RBC diluting pipette and pipette filler Blood is drawn up to the o.5 mark THE diluting fluid is then drawn up to the 101 mark ( this gives a dilution of 1/200) Mix thoroughly for at least 2-3 minutes

9. RED BLOOD CELL DILUTING FLUID formal citrate -sodium citrate: 3.0gm -formaldehyde/formalin: 1.0ml -distilled water: 100ML * Any isotonic solution are acceptable such as normal saline

10. Counting the rbc or/and platelet prepare the counting chamber and the cover slip charge one of the counting areas the fluid will fill up the space by capillary action, DO NOT OVERFILL allow a few minutes for the cells to settle

11. TRIPLE RULING METHOD - to avoid duplication in counting cell that touch the lines triple ruling is used cells that touch top and left central lines are counted. Cells that touch bottom and right central lines are not counted

12. RBC RBC RBC RBC CALCULATE RBC one in the center & four of the corner RBC

13. TOTAL WHITE CELL COUNT & PLATELET WBC DILUTING FLUID TURKS’ SOLUTION -glacial acetic acid: 2.0ml -1% gentian violet: 1.0ml -Distilled water: 97ml

14. PLATELET COUNT Platelet diluting fluid rees ecker -Sodium citrate 3.8 gm -Formalin: 0.2 ml -Brilliant cresyl blue: 0.1 gm -Distilled water: 100 ml

15. PROCEDURE WBC COUNT Filling the pipette using the wbc pipette Draw blood to 0.5 mark and follow up by diluting fluid up to 11 mark (this give a dilution of 1/20)

16. Counting the white blood cell locate the 4 large corner square, marked w under 4x objective Count using the triple rulling method

17. WBC WBC WBC WBC Count the 4 large square at the grid corner

18. CALCULATION RED BLOOD CELL: (a + b + C + d + e) /5 * (200) * (0.004mm³/ cm³) :_________ cm³ cell per cubic cm White blood cell: (a + B + c + D) /4 * (20) * (0.1 mm³ cm³ ) :__________X cm³ cell per cubic cm PLATELET COUNT: (All small squares) / 400 * (20) * (0.00025MM³ / 2.5 x :_______________ / 2.5 x cell per cubic cm  

19. Precaution during cell counting Clean hemocytometer , coverslip and pipette Diluting fluid need to be contaminant-free Adequate mixing to prevent clumping-platelet Presence of air bubbles in pipette Excess blood from the pipette tips

20. THANK YOU FOR YOUR ATTENTION