LBC kit
includes:-
1.slides
coated with adhesives
2. Cell separator
solution.
3.slide
holder and bush
4.
Disposable graduated pipettes
5.sample
container with preservative
6. Cyto brush with detachable head with bristles.
Sample is
preserved in the fixative which is pre-filled by manufacturer.
Smear
preparation for PAP-LBC:-
1. Sample
received
2. 4 ml of
cell separator solution is taken in graduated conical centrifuge tube.
3. 7ml of
sample (well mixed) is poured into the tube by making low angle as they should
not mix-up before centrifugation.
The total volume will be 11 ml.
4. Spin the
tube at 1500-2000 rpm for 10-15 minutes.
5. Discard
the supernatant and resuspend the sediment
6. Place
the slide in slide holder and lock the bush by pushing lightly and rotating.
7. Put
100ul D/w and 100ul resuspend sediment // 200ul D/W +50ul of sample.
{The quantity
of D/W and sample can be changed according to the thickness required for the
smear.}
8. Tap
slightly to evenly distribute the sample and put it in the centrifuge specially
designed for LBC at 2000 rpm for 2 minutes.
9. Remove the slide and wash in running tape
water.
10. Fix the
slide at least for 2 minutes in ethyl alcohol or isopropyl alcohol.
11. Remove
the slide and keep in rack for complete drying.
Staining:-
Rapid PAP-
staining kit contains
1. Nuclear
stain
2.
Cytoplasmic stain
3. Xylene
4. Cover
slip
5. DPX mountant
Steps:-
1. Put the
dried slides in the nuclear stain jar for 3-4minutes .
2. Wash in
running tape water.
3. Put the
slides in nuclear stain jar for 3-4 minutes .
4. Wash in
running tape water.
5. 1-2 dip
in alcohol for dehydration
6. Allow it
to dry in air
7. Dip in
xylene and mount the smear using DPX.
and Observe
under microscope .
Result:-
Nucleus:- blue
Cytoplasm: -
varying shades of pink, blue, yellow green -grey
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