Differential Leukocyte count

Differential leukocyte count

The blood cells have formed element and one of them is white blood cells and white blood cells are classified on garnulocytes and agranulocytes according to presence and absence of granules on cells.

Granulocyte include neutrophil, eusinophil and basophil and

Agranulocyte include lymphocyte and monocyte

To count these types of cells we should perform differential leukocyte count.

Material required

Blood sample
Microscope
Slides
Wright’s stain solution
Distilled water
Immersion oil
Standing rack
Pipette
Blow pipe
Glass rod
Cotton

Procedure

1. Preparation of blood smear
2. Staining of blood smear
3. Identification and counting of different types of leukocytes.

Preparation of blood smear

Ø Take two grease free, clean, dry with smooth edges slide and keep horizontally one slide on the table.
Ø Put One medium size of drop of blood on the left side of the slide(smearing slide)
Ø Placed another slide at middle of the horizontal slide at 45 degree and pull backward touching the blood.
Ø When the blood is spread in edges of spreader slide then push the slide toward forward at 45 angle degree with continuous pressure.
Ø Leave prepared slide for air dry.

Staining of blood smear

Ø Place the slide on the staining rack and put appropriate amount of wright’s stain solution and leave for 3-4 minute.
Ø Add equal amount of distilled amount of on the slide and blow with blow pipette.
Ø When bronze like scum is appeared wash it with running water and leave it for air dry.

Identification and counting of leukocyte

Ø Keep the slide under microscope and see the staining quality at 10X objective lens.
Ø Add immersion oil on the slide and placed on 100X objective lens and start counting of through parallel strip method.
Ø A total 100 leukocyte well be counted and number of different types of leukocytes will be expressed as percentage out of 100 cells.