1. White Blood Cells Count Using Hemocytometer
Salman bin Abdulaziz University
College of Pharmacy
Clinical Pharmacy Department
Physiology-I
(Practical Part)
White Blood Cells Count Using Hemocytometer
9/17/2018

2. White Blood Cells - WBCs
white blood cell, also called leukocyte ,  a cellular component of the blood that lacks hemoglobin, has a nucleus, is capable of defends the body against infection and disease by ingesting foreign materials and cellular debris, by destroying infectious agents and cancer cells, or by producing antibodies.
The normal range of leukocyte is 4, ,000/cubic millimeter.
Red blood cells are confined to the bloodstream and functions in the blood. White blood cells, by contrast, are able to slip into and out of the blood vessels and go to tissues (by process called diapedesis).
WBCs Disorders:
1. Leukocytosis: is a high white blood cell count
Microbes, anesthesia or surgery
2. Leukopenia is low white blood cell count
radiation, shock or chemotherapy.
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4. WBCs Disorders: 1. Leukocytosis is a high white blood cell count
Microbes, anesthesia or surgery.
2. Leukopenia is low white blood cell count
radiation, shock or chemotherapy.
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5. White Blood Cells Types
WBC’s are divided into two categories: Granulocytes, and Agranulocytes.
Granulocytes contain granules within the cytoplasm and include: Neutrophils, Eosinophils, and Basophils.
Agranulocytes contain very little, if any, granules and include: Lymphocytes and Monocytes.
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6. White Blood Cells Types
Neutrophils (Granulocyte)
Neutrophils have a multilobed nucleus (3-5lobes) and very fine granules
Function: Release lysozymes which destroy/digest bacteria
Neutrophils are produced by bone marrow, they fight disease by migrating to the point of infection, absorbing bacteria, and destroying them (phagocytosis)
Neutrophil (purple) migrating through tissue to engulf bacteria through phagocytosis.
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7. Eosinophils (Granulocyte)
Nucleus with 2 or 3 lobes connected by a thin strand
Function: Attack parasitic worms and play an important role during allergies and asthma.
albendazole
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8. Basophils (Granulocyte)
The rarest of the WBCs, have S shaped nucleus contain large histamine-containing granules that stain dark blue.
Function: Basophils are involved in inflammatory and allergy by releasing heparin, histamine & serotonin.
the rarest of the WBCs, have S shaped nucleus contain
large histamine-containing granules that stain dark blue.
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9. Lymphocyte (Agranulocyte)
Dark, oval to round nucleus
Cytoplasm sky blue in color
Function:
B cells:
Destroy bacteria and their toxins
Turn into plasma cells that produces antibodies
T cells
Attack viruses, fungi, transplanted organs, cancer cells & some bacteria.
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10. Monocyte (Agranulocyte)
Monocytes are the largest of the WBCs. Except for their more abundant cytoplasm and indented (kidney like) nucleus, they resemble large lymphocytes. When they migrate into the tissues, they change into macrophages. Macrophages are very important in fighting chronic infections, such as tuberculosis.
Function: Macrophages absorb disease-producing materials, such as bacteria through phagocytosis.
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12. White Blood Cell (WBC) Count
Expressed as: Number of white blood cells per a unit volume
(x 103 / cu.mm.)
(x 103 / mm3.)
(x 103 / µL.)
(x 109 / L.)

13. WBC Count (Cont.) Reagent WBC diluting fluids 1. Turk’s solution
Specific Equipment
White pipette
(Thoma type)
2. Rubber pipe
3. Plastic syringe
4. Pipette vibrator
5. Hemocytometer & cover slip (Counting chamber)
6. Microscope
7. Counter (hand tally)
8. Report form
Reagent
WBC diluting fluids
1. Turk’s solution
Glacial acetic acid
Gential violet
Distilled water
2. Acetic acid (2%)
3. Hydrochloric acid (1% )

14. WBC Count (Cont.) Procedure 1. Mix the blood sample thoroughly.
2. Fill blood into white pipette at mark.
3. Fill reagent add up into the pipette
to 11 mark.
4. Shake the pipette on the vibrator for 1 min.
5. Discarded the first 3-4 drops.
6. Fill in the hemacytometer nicely.
7. Allow WBC to set down for 2-3 min.
8. Count 4 white squares under microscope (x100 lens).
9. Calculate the WBC concentration.

15. WBC Count (Cont.) High 0.1 mm. 1 mm. W W 1 mm. R R 1 mm. R R R W W

16. Hemacytometer counting chamber (WBCs)
Hemacytometer counting chamber (WBCs). Areas marked A, B, C, and D are used to count white blood cells.

17. WBC Count (Cont.) W
1 mm.
High 0.1 mm.
1 mm.

18. WBC Count (Contd)
Top
Left

19. WBC Count Calculation 1. Volume of 1 white square = 1 x 1 x 0.1 cu.mm.
2. Volume of 4 white square = 0.1 x 4 cu.mm.
= 0.4 cu.mm.
3. In 0.4 cu.mm. the WBC count = N (counted No.)
4. In 1 cu.mm. the WBC count = N x 1 / 0.4
= N x 2.5
5. The dilution for WBC = 10/0.5
= 20
6. The final WBC count = N x 2.5 x 20
= 50 x N (/cu.mm.)

20. WBC Count (Cont.) Calculation Normal range = 4,800 - 10,000/ cu.mm.
White cell count = number of cells counted (N)
x volume factor (=2.5)
x dilution factor (=20)
= N x 50
Normal range = 4, ,000/ cu.mm.
x 103 / cu.mm.

21. Do You Have Any Questions?
Thank You
For
Your Attention
Do You Have Any Questions?
9/17/2018