For
the laboratory diagnosis of the samples l both automation analysis and
manual:
3.9.1 For the
staining of thin film:
3.9.2 Method Used: Leishmann staining
method
This
is based on the fact that leishmann stain is a type of romanowsky stain
which contains eosion Y which is a cidic
anionic dye and azure B and other thiazine
dyes which are basic
cationic dyes. When diluted in buffered water ionization
occurs eosion stains the basic components of blood cells, e.g
hemoglobin stains pink-red, and
the granules of eosinophils stain
orange-red . azure B and other methylene
blue derived dyes, stain the acidic components of the cells. Nucleic acids and nucleoprotein stain various shades
of mauve-purple and violet, the granules of basophils stain dark blue-violet,
and the cytoplasm of monocytes and
lymphocytes stains blue or
blue-grey . the staining
reactions fo romanowsky stained
pre PH dependent which is why the
stains are diluted in buffered water of specific PH
(cheesbrough , 2000)
Procedure
1.
using
clean grese –free slides placed a drop of blood in the center of a given slide
about 1 cm from the end
2.
Without delay a spreader was placed in front of the drop at an angel
of about 45o C to the
slide.
3.
Moved
the spreader slightly backward so
as to make contact with the drop
4.
The
drop was spread out quickly along
the line
of contact
5.
With the steady movement of the
hand, the drop was spread
along the slide
6.
Allowed
to air-dry, labeled and fixed without
delay using methanol for 1 minutes
7.
After 2 minutes of fixation, stained with leishmann stain without over-flooding
8.
Added twice
the volume of PH 6.8 buffered water ensuring that it was well mixed
9.
Allowed
to stain for 10minutes
10.
Washed of the stain with tap water
11.
Wiped
the back of the slide gently with
cotton wool and place in the draining
rack for the slides to dry completely.
12.
Counted
the cells microscopically using oil
immersion objectives