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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