Equipment and materials
The following has been prepared for you
You will be provided with a pre-prepared slide for comparison.
h
You have been provided with
Diff Quick stain
Cover slips
Immersion oil
Staining/drying racks
Blotting paper
Stopwatch
Method
This scheme is a guide only; slides should be inspected during staining and the procedure adjusted if necessary. Pipette reagents onto the preparation to just cover it; there is no need to waste quantities of reagents down the sink. Never place a wet mount on a microscope stage without cleaning and drying the back of the slide first.
Staining of prepared blood films
a) Place the slide on a staining rack.
b) Drop fixative on the slide, covering the blood film, blow gently across the surface for 5 seconds. Tip off fixative.
c) Drop Speedy Diff A on the slide covering the blood film, blow gently across the surface for 5 seconds. Tip off the stain.
d) Gently wash the slide over with distilled water, twice.
e) Drop Speedy Diff B on the slide covering the blood film, blow gently across the surface for 5 seconds. Tip off the stain.
f) Gently wash the slide over with distilled water, five times.
g) Stand upright on blotting paper to drain and allow to dry.
h) Clean the back of the slide and examine under oil immersion. Patience is required to scan the slide systematically for all the white cell types. (Non-ideally: granulocytes tend to aggregate at the edge of the film if they have been damaged during spreading).
2) Identify the different cell types by size and staining reaction and make large annotated High Power diagrams of your observations. Red cells can be used as a size guide. Count and record the leukocytes you see.
3) Write blood film reports for your samples.
