1. Preparation, staining and examination of blood film
M. Sc. Program
541 CLS
Lab-4-

2. Blood film Blood film should be made on clean glass slides.
Films may be spread by hand or by automated slid-spreader.
Blood film prepared from fresh blood (EDTA).

3. STEPS FOR BLOOD FILM

4. The shape of blood film

5. Staining of blood films
Romanowsky stains are universally employed for routine staining of blood films.
It depends on two components, namely, azure B(trimethylthionin) and eosine Y(tetrabromo-fluorescein).
The original Romanowsky combination was polychrome methylene blue and eosine.
The presence of basic grouping on the haemoglobin molecule results in its affinity for acidic dyes and its staining by eosin. The nucleic acids uptake of the basic dye azure B.
Granules in cytoplasm of neutrophil are weekly stained by the azure complexes( eosinophilic granules-alkalin grouping which stains with acidic component of dye wherase basophilic granules which has an affinity for the basic component of dye).

6. Types of Romanowsky stains
Leishman’s stain
May-Grunwald stain
Jenner’s stain
Giemsa’s stain
Jenner-Giemsa
May-Grunwald-Giemsa

7. Red cell morphology
In health, the red cells have round smooth contours and have diameters of 6.0 to 8.5µm.
Abnormalerythropoiesis:
Anisocytosis (size: micro or macro).
Poikilocytosis (shape)
Basophilic stippling
Acanthocytosis, target cell and sickle cell.

8. Erythrocyte inclusion (Howell-jolly bodies, Pappenheimer bodies
According HB: hypochromatic and hyperchromatic.
Spherocytosis, elliptocytosis (cell membrane of red cell).