WBC manual count using
hemocytometer
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Introduction


WBCs count is the count of leukocytes
in a volume of blood
Expressed as WBCs/mm³
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Material
1.
2.
3.
4.
5.
Blood sample (EDTA anticoagulated
blood or capillary blood)
WBCs diluting pipette
Diluting solution
Hemocytometer
microscope
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The Hemacytometer


contains 2
Neubauer counting
chamber
Each chamber
contains:


*4 WBC counting
squares
*Each contains 16
squares
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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.
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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
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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.
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Count and calculate:
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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
9
Calculation

Number of cells/mm³=
counted cells in 4 large square /4 x
diluting factor x
volume correction factor.
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Calculation

The dilution factor=
total volume/sample volume
= 11-1/0.5 =20
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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

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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
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Example:


If total number of WBCs in 4 squares is
120
Then the number of WBCs in 1mm³=
120 /4 x 200 =6000
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Normal values of WBCs:







Newborn 9000-30000 cell/mm³
Adult 4000-11000 cell/mm³
Neutrophil 62%
Eosinophil 2.3%
Basophil 0.4%
Monocytes 5.3%
Lymphocyte 30%
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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

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Significance of the test

Leukopenia








Result from X-ray therapy
Alcoholism
Antibiotic therapy
Typhoid infection
Measls
Infectious hepatitis
TB
And cirrhosis of the liver
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THANKS
Thanks
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