Differential Count of WBC
Differential count of WBC involves determining the concentration of each type of leucocyte. There are 5 different types of leucocytes; neutrophil, eosinophil, basophil, monocyte, lymphocyte. Differential count of WBC is done by preparing a blood smear and staining slide with suitable stain and counting individual number of leucocyte.
Materials Required:
- Glass slide
- Stain; Giemsa stain, Leishman stain or Wright stain
- Microscope
- Hand counter
- Micropipette
- Uncoagulated blood sample
Procedure:
- A drop of blood is placed on clean, grease free slide with help of pipette
- Blood is spreaded with another slide at 45° angle to make uniform smear.
- Glass slides are allowed to dry for some time after formation of smear
- Glass slides are then stained using any one of stain; Leishman stain, giemsa stain
- Staining are done by placing slide on staining rack. Blood smear is covered with Leishman stain for a minute.
- Equal quantity of buffer is added and mixed by blowing. This mixture is allowed to remain on slide for 10 minutes.
- It is then washed in running water. Slides are allowed to dry in air for some time and observed under microscope under 10X. then slides are examined under 100 x under oil immersion
- A corner area is choosen and cells are counted and recorded on hand counter. Cells are counted till 100 cells are observed and percentage of each leukocyte is determined.
Differential leucocyte percentage in different animals
|
|
Cattle |
Buffalo |
Horse |
Goat |
Sheep |
Pig |
Dog |
Cat |
|
Neutrophil |
29.10 |
28.70 |
62 |
40 |
41 |
39 |
62.80 |
44.82 |
|
Eosinophil |
9.87 |
2.90 |
4 |
2.50 |
5.10 |
4.50 |
2-14 |
2-11 |
|
Lymphocyte |
51.40 |
60.60 |
20 |
48 |
50 |
52 |
10-28 |
15-44 |
|
Monocyte |
8.32 |
7.80 |
8 |
2 |
3 |
3.30 |
3-9 |
0.5-0.7 |
|
Basophil |
0-1 |
0.00 |
2 |
1.60 |
0.03 |
1.20 |
0-2 |
0-0.05 |