Determination of Differential Count of W.B.C

 

Determination of Differential Count of W.B.C

 

Apparatus Required: Microscope, Glass Slides Spirit and pricking Needle Leishman’s stain, Distilled water, staining Rack, Cotton, Lancet.

 

Leishman’s Stain: It contains both basic (Methylene Blue) and acidic (eosin) stain dissolved in Acetone free methyl Alcohol. The methyl Alcohol fixes the smear to the slide. It is prepared by adding 1.5 g of dry stain to 1000 ml. of acetone free methyl alcohol. The stain improves on exposing to sunlight.

 

Method: Prepare an ideal peripheral blood smear which is thin, smooth & homogenous.

 

                No-windows & serration it is usually tongue shaped these are the characters of a good and ideal smear. 2-3 or the glass slide PBS- Dry the smear.

 

Staining: I put the slide on the staining rack & pour sufficiently the leishman’s stain & wait for 2-3 mts (fixition stage). Note the number of drops used. NB: The stain should not be dry on the slide.

 

II. Add double the amount of distilled water over the slide & below the contents gently so that it mixes properly & wait for 8-10mm. (staining takes at the end of 8-10 min.)

 

III. Discard the diluted stain & Rinse gently with tap water to remove excess stain (ideal pinkish blue).

 

IV) Dry the smear thoroughly & then put a drop of cedar wood oil or liquid paraffin & focus under immersion lens.

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