Preparation, staining and examination of blood film

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Preparation, staining and examination of blood
film

• M. Sc. Program
• 541 CLS
• Lab-4-

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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).

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STEPS FOR BLOOD FILM
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The shape of blood film
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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-fluore
  scein).
• 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).

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Types of Romanowsky stains

• Leishmans stain
• May-Grunwald stain
• Jenners stain
• Giemsas stain
• Jenner-Giemsa
• May-Grunwald-Giemsa

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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.

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