Preparation of blood films for malaria parasites
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MALARIA DIAGNOSIS WORKSHOP
29 JUNE – 2 JULY 2010
PREPARATION OF BLOOD
FILM FOR MALARIA
PARASITES
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Two types of blood film for malaria parasites
Thick Blood Smear – use to determine if parasite is present.
Thin Blood Smear – use to confirm the Plasmodiumspecies present
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Before making blood film…
Wear gloves prior to taking blood samples or handling blood.
Avoid getting blood, including dry blood from films, on your fingers or hands.
Cover cuts on your hands with waterproof dressing.
Avoid accidentally pricking yourself when handling sharp instruments
Wash your hands with soap and water after you finish a job.
If you get blood on your skin, quickly wipe it off with a cotton swab dampened with alcohol; then, wash the affected area with soap and water as soon as
possible.
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Materials for Preparation of Malaria
Smears
Clean and wrapped slides
Sterile lancets
70% ethanol and water
Absorbent cotton wool
Surgical gloves
Lint-free cotton cloth
Slide box or cover to exclude flies and dust
Record form or register
Soft lead pencil; ball-point pen
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PROCEDURES:
1. RECORD PATIENT DETAILS ON A REGISTER
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Get ready clean
microscopic slide
glass slides of
superior’ quality, with
frosted end.
The frosted end
should be used to
label the slide.
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PREPARATION OF BLOOD FILM FROM
FINGERPRICK
2. Wear glove and hold
the patient’s left hand
with palm facing upwards.
Select third finger from
the thumb
The thumb should never
be used for adults or
children.
Note : Good films may be prepared
directly from fingerprick blood,
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Clean the finger with
a piece of cotton wool
lightly soaked in 70%
ethanol.
Dry the finger with a
clean cotton cloth,
using firm strokes to
stimulate blood
circulation.
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For infant – use the
toe
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4. Use sterile lancet to
prick the finger
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5. Apply gentle pressure
to the finger to allow
the blood to ooze out
6. Wipe off the first drop
7. Apply further gentle
pressure for more
blood
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8. Place blood on clean
glass slide to make
thick and thin blood
film on a slide
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Place the spreader
slide in contact with
the drop of blood at
an angle..
Let blood spread
along the edge of the
spreader slide.
gently push towards
the other end of the
slide
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Thin Blood Film
Angle too flat
> film too long
Angle too steep
> film too short
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Thin Film Good preparation - feathered end of
the film should be centrally located on the slide with free margins on both sides,
when properly prepared, it will be only one cell layer thick at this end.
Badly prepared smears can cause presence of streaks - as a result of chipped spreader
holes in the film indicate faulty preparation and dirty or greasy slides, respectively.
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Combination of thick and thin blood on the same slide
OK
TOO MUCH BLOOD
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Thick blood film – spread the drop of blood using a corner of the spreader.
Spread the drop in one circular direction to make even thick film, size of a 1 cm diameter. Do not stir the blood.
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Preparation of thick blood film using …….
Plastic pipette
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Using wooden stick
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Good thick blood film
• The right thickness,
• You should be able to
read the newspaper,
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Avoid doing these
Thick film too thickIt should be round
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Labeling
With frosted end
use a pencil
Without frosted end
use diamond pencil
Do not label on the
blood film
Do not use a ball pen
Do not use a paper
label
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Drying slides
Incubator 37oC
Leave standing on a
rack on lab bench
In a slide tray with the
cover open
Hair dryer?
A fan?
Do not leave on the bench in a laboratory
unattended because it will be eaten by insects.
Slides must be dried completely before they
are packed for storage or transport.
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Venous blood can be
used instead of from
finger prick
Use vacutainers with
anticoagulant (EDTA)
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For preparing thin and
thick films use a glass
capillary to drop the
ETDA-blood.
plastic pipette ? Yes
provided no air bubles
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The use of anticoagulated (EDTA) blood may cause the following :
smears require longer time to dry
thick smears tend to flake from the slide
stain quality is affected,
stippling of infected RBC’s may not be visible
parasite forms may be distorted and may lyse
RBC’s may become crenated and look fimbrinated
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Prepare smears as soon as possible after
collecting venous blood (less than 2
hours after collection) to avoid:
Changes in parasite morphology
Changes in the RBC morphology
(these effects can compromise
correct identification of species)
Staining characteristics
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YouThank
For your attention