Lab 4 : White Blood Cell Count Objectives

• 1. Perform routine white blood cell count by using

hemocytometer

• 2. Know how to calculate the number of white blood

cell

• 3. Interpret and correlate the results with various

abnormalities

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 Wbc count: Procedure
1. Mix the blood sample thoroughly.
2. Fill blood into white pipette at 0.5 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 (x400).
9. Calculate the WBC concentration.

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• Using dilution pipette with the white mixer, draw
blood up to the 0.5 mark.
• Dab with piece of paper towel if needed to adjust
volume.

3
• Fill the pipette the rest the 11 mark with WBC diluent.
• Shake well to mix with the hose end sealed with your finger.
WBC diluent :
- 10 mg crystal violet
- 1.0 ml glacial acetic acid
- 100 ml with d H20

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• Empty ~2-3 drops of pipette into waste container

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• Add a small amount of the diluted blood to just fill
the first chamber of the hemacytometer.

• It should flow in to fill the

chamber by capillary action.
• Do not over fill.
• Let the preparation sit for a minute
(for cells to settle).

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• To improve your skill, repeat the dilution a second time and
fill the second chamber.
• After completing the counts of each, compare the numbers
you have generated.

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A B
1 2

3 4

C D 8
9
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 Calculation
• Blood is drawn to the 0.5 mark and diluted to the 11
mark with WBC diluting fluid.
• All the blood is washed into the bulb of the pipet
(which has a volume of 10).
• Therefore, 0.5 volumes of blood are contained in 10
volumes of diluting fluid.
• The resulting dilution is 1:20.

11
 1 mm.

W W 1 mm.

R R
R High 0.1 mm.
R R

W W

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 Calculation
1. Volume of 1 white square = 1 x 1 x 0.1 cu.mm.
= 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 cells
4. In 1 cu.mm. the WBC count =N cells
0.4
5. The dilution for WBC = 1:20
6. The final WBC count = N x 20
0.4 cells/cu.mm.
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 • For example

Calculate the average number of WBCs per chamber:

Calculate the number of WBCs per cubic mm:

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