1. WBC manual count using hemocytometer

2. Introduction
WBCs count is the count of leukocytes in a volume of blood
Expressed as WBCs/mm³

3. Material Blood sample (EDTA anticoagulated blood or capillary blood)
WBCs diluting pipette
Diluting solution
Hemocytometer
microscope

4. The Hemacytometer contains 2 Neubauer counting chamber
Each chamber contains:
*4 WBC counting squares
*Each contains 16 squares

5. Methodology
draw blood up to 0.5 marks on WBCs pipette and complete to 11 with WBCs diluting solution(Turk‘s solution).
Mix for 2-3 minute.

6. Turk's solution
Türk's solution is a composed of a stain (Gentian violet) and 1-2% acetic acid. The solution destroys red blood cells within a blood sample, and stains the nuclei of the white blood cells, making them easier to see and count. Turk's Solution is intended for use in counting leukocytes in a defined volume of blood. Erythrocytes are hemolyzed while leukocytes are stained for easy visualization.
Dr.Sajeda Al-Chalaby

7. Charge hemacytometer
Discard the first 4-5 drops
Place tip of the pipette at the edge central platform
Then let the hemacytometer to stand on the bench for 3-5 minute.

8. Count and calculate:

9. Turk's solution
Türk's solution is a composed of a stain (Gentian violet) and 1-2% acetic acid. The solution destroys red blood cells within a blood sample, and stains the nuclei of the white blood cells, making them easier to see and count. Turk's Solution is intended for use in counting leukocytes in a defined volume of blood. Erythrocytes are hemolyzed while leukocytes are stained for easy visualization.
Dr.Sajeda Al-Chalaby

10. Calculation Number of cells/mm³= counted cells in 4 large square/ 4 x
diluting factor x
volume correction factor.

11. Calculation The dilution factor= total volume/sample volume
= 11-1/0.5 =20

12. Calculation counted volume = The total volume of the 1 large square=
= (width x length x depth )
= (1mm x 1mm x 1/10 mm) =1/10

13. Calculation Number of cells/mm³= counted cells in 1 large square x
diluting factor x
volume correction factor.
Number of cells/mm³= n x20 x 10
= n x 200

14. Example: If total number of WBCs in 4 squares is 120
Then the number of WBCs in 1mm³=
120/ 4 x 200 =6000

15. Normal values of WBCs: Newborn 9000-30000 cell/mm³
Adult cell/mm³
Neutrophil 62%
Eosinophil 2.3%
Basophil %
Monocytes 5.3%
Lymphocyte 30%

16. Significance of the test
Leukocytosis
Occur as an indicator of body defense against foreign materials (bacteria, parasites, toxins)
Metabolic disorders
Chemical and drug poisoning
Acute hemorrhage

17. Significance of the test
Leukopenia
Result from X-ray therapy
Alcoholism
Antibiotic therapy
Typhoid infection
Measls
Infectious hepatitis TB
And cirrhosis of the liver

18. THANKS
Thanks