Preparation, staining and examination of blood film

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Presentation transcript:

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

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).

STEPS FOR BLOOD FILM

The shape of blood film

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).

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

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.

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