This document provides instructions for performing a blood cell count using a hemocytometer. It explains that a hemocytometer is used to count blood cells and involves diluting a blood sample then counting the cells under a microscope. The key steps are filling pipettes with diluted blood and diluting fluid, charging the counting chamber, allowing cells to settle, and counting cells that touch the ruled lines. Calculations are provided to determine total red blood cell, white blood cell and platelet counts based on the number of cells counted. Precautions like cleaning equipment and ensuring no clumping or bubbles are also outlined.

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
)

7. 2. BLOOD DILUTING PIPETTES
RBC
WBC

8. 3. MICROSCOPE

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

10. 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

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

12. 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

14. RB
C
RB
C
RB
C
RB
C
CALCULATE
RBC one in
the center &
four of the
corner
RB
C

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

17. 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

18. 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)

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

20. WB
C
WBC
WBC
WBC
Count the 4
large square
at the grid
corner

21. CALCULATION
 RED BLOOD CELL: (a + b + C + d + e) /5 * (200) *
(0.004mm³/4 𝑥 10−6
cm³)
:_________ 4 𝑋 10−6 cm³ cell per cubic cm
 White blood cell: (a + B + c + D) /4 * (20) * (0.1 mm³ /10−4 cm³ )
:__________X 10−4 cm³ cell per cubic cm
 PLATELET COUNT: (All small squares) / 400 * (20) * (0.00025MM³
/ 2.5 x 10−7
𝐶𝑀³)
 :_______________ / 2.5 x 10−7
𝐶𝑀³) cell per cubic cm

22. 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

23. THANK YOU FOR YOUR ATTENTION