EXAMINATION OF

BLOOD
Done by: Miss Nada
 Examination of blood
 Examining the specimen of blood

microscopically for:

1. Plasmodium spp. ( Malaria parasites).

2. Trypanosoma spp.

3. Microfilaria spp.
 What is malaria?
 Malaria is a vector-borne infectious
disease caused by single-celled
protozoan parasites of the genus
Plasmodium.
 Malaria is transmitted from person to
person by the bite of an infected
female Anopheles mosquitoes.
 What is malaria?
 Infectioncan also occur by
transfusion of infected donor
blood, by injection through the
use of needles contaminated with
infected blood.
 Plasmodium species
1. Plasmodia falciparum,
2. Plasmodia vivax,
3. Plasmodia malariae and
4. Plasmodia ovale.
The most dangerous of the four is P.falciparum.
 Laboratory diagnosis:
 Methods in Parasitology:
 Preparing thin and thick blood films with
capillary or venous blood.
 Materials for finger:

• Disinfectant
• Swabs
• Microscope slides (cleaned with alcohol)
• Sterile lancets
• Special slide as spreader
• Gloves
 Preparing thin and thick blood
films

 Prepare the
microscope slides
 Preparing thin and thick blood
films
(capillary blood)
With the patient‘s left hand,
palm upwards, select the
third finger. (The big toe
can be used with children.
The thumb should never be
used for adults or children).
Use cotton wool lightly
soaked in alcohol to clean
the finger. Let finger
air-dry.
 Preparing thin and thick blood
films

(capillary blood)
With a sterile lancet
puncture the ball of
the finger using a
quick rolling action.
 Preparing thin and thick blood
films
By applying gentle
pressure to the finger
express the first drop
of blood and wipe it
away with dry cotton
wool.
Make sure no strands
of cotton remain on
the finger.
 Preparing thin and thick blood
films
Working quickly with
capillary blood and
handling clean slides only
by the edges, collect the
blood as follows:
Apply gentle pressure to
the finger and collect a
single small drop of blood
about the size ● on the
end of the slide. This is
for thin film.
 Venous blood can
be used instead of
capillary blood.

 Usevacutainers
with anticoagulant
(EDTA)
For preparing thin
and thick films use a
glass capillary tube
or automated
pipette to drop the
ETDA blood.
Do not use a plastic
pipette!
 Preparing thin and thick blood
films
1. Hold the spreader at 30-45degree angle
against the surface of the slide.
2. Move the spreader back to touch the
drop of blood and allow the blood to
extend along the edge of the spreader.
3. Push the spreader with a steady hand
across the slide.
4. Air dry the film.
 Preparing thin and thick blood
films
Possible mistakes:

 This is wrong!

 This is correct!
 Preparing thin and thick blood
films
 Observe right angle:
Angle too flat film
too long

Angle too steep film

too short
 Preparing thin and thick blood
films
 Pressureon
spreader:
too strong waves

Air bubble holes

 Preparing thin and thick blood
films
1. A large drop of blood is taken on the
centre of a clean labeled slide ( a drop of
finger puncture or from well mixed EDTA.
tube of blood by capillary tube or pipette).
2. With an other slide corner spread the
drop over 1/2 an inch square area.
3. Air dry the film.
 Preparing thin and thick blood
films

Thick films
Check for the right
thickness:
You should be able
to read the
newspaper!
 Preparing thin and thick blood
films
Additional mistakes
• Thick film too thick

• Thick film should be

round!
 Preparing thin and thick blood
films

These slides look

o.k.
 Preparing thin and thick blood
films

FIXING OF THIN BLOOD FILMS:

* With absolute methanol, by immersing in a

container of absolute methanol for 2 minutes.

* Methanol containing water must not be used

to fix blood films.
 Preparing thin and thick blood
films
Staining thin blood film (Leishman stain):
☻ Cover the blood film with undiluted
Leishman stain, but do not flood the
slide, allow to stain for 2 min.
☻ Add twice the volume buffered water.
☻ Mix water and stain by bowling on the dilute
stain.
☻ Allow to stain for 10 minutes.
☻ Wash off the stain with tap water, wipe the
back of the slide clean and stand it in a rack
for the smear to dry.
 Preparing thin and thick blood
films
  STAINING THICK BLOOD FILMS (Giemsa Staining):
 
☻ Dilute the Giemsa stain as required:
10% solution for 10 minutes staining:
Measure 150 ml of buffered water in a 200 ml
cylinder. Add 15 ml of Giemsa stain and mix gently.
☻ Cover the blood film Giemsa stain.
☻ Allow to stain for 10 minutes.
☻ Wash the stain using clean water.
☻ Wipe the back of the slide clean and stand it in a
rack for the smear to dry.
microscopic examination of blood

PLASMODIUM FALCIPARUM

PLASMODIUM VIVAX
microscopic examination of blood

PLASMODIUM MALARIAE

PLASMODIUM OVALE
 microscopic examination of blood
 For Malaria microscopic diagnosis we
looking for:
2. Appearnce of RBC (characteristics)
3. Ring forms
4. Trophozoites
5. Schizonts
6. Gametocytes (macro & micro
gametocyte)
 RBC Plasmodium
appearance: vivax
• Size: larger than mature RBC
• Color: Pale
• Shape: Round
• Cytoplasmic inclusions: Schuffner’s
dots present
• usually no more than one parasite is
observed within a single enlarged red
blood cell
Plasmodium vivax
 Plasmodium falciparum
 RBC appearance

• Size: Mature RBC

• Color: Normal
• Shape: Round
• Cytoplasmic inclusions: Maurer’s clefts
may be present in late trophoziots
Plasmodium falciparum
Ring Form Schizont stage
 Plasmodium falciparum
Gametocytes of P. falciprum
Macro-gametocyte Microgametocyte

Size: larger than RBC Size: larger than RBC

Shape: crescent –sharply Shape: kidney-bluntly round
pointed end end
Cytoplasm: dark Cytoplasm: riddish
Chromatine: compact mass Chromatine: fine granules
near the center scattered throughout
Pigment: black granules Pigment: dark granules
round the center. throughout
 Plasmodium ovale
 RBC appearance

 Size:larger than mature RBC

 Color: pale
 Shape: oval
 Cytoplasmic inclusions: James’ dots
 Plasmodium malaria
 RBC appearance

 Size:smaller and older RBC

 Color: Normal
 Shape: Round
 Cytoplasmic inclusions: None
Thank you