UNIVERSITI TEKNOLOGI MARA

(BERTAM, PULAU PINANG)

Faculty of Health Sciences
Diploma in Medical Laboratory
Technology
______________________________________

CSI 153
CLINICAL PARASITOLOGY
PRACTICAL 5a & 5b:
Preparation of thick and thin blood smear & Giemsa
staining of thin and thick blood smear

NAME : NURUL FATINAJIHAH BINTI ZULIKIPLI

STUDENT ID : 2016625096

IC NUMBER : 981012115026

LECTURER’S NAME : MADAM NURHIDAYAH BINTI

AB. RAHIM
INTRODUCTION

Thick and thin blood smear was used to determine wether we have malaria or not
and it was done in the laboratory. If one test is negative and no parasites are found, we will
have repeated blood smears every 8 hours for a couple of days to confirm that there is no
malaria infection in the patient body.Blood smears are taken most often from a finger prick.
Thick and thin blood smears will let doctors know the percentage of red blood cells that are
infected (parasite density) and what type of parasites are present in the body.We have to do
the preparation of thick and thin blood smear then continue with Giemsa staining of thick and
thin blood smear.

For the preparation of thick blood smear, three small drops of blood were placed on a
clean glass slide.Thick blood smear was very important as it is useful for detecting the
presence of parasites.This is because we examine a larger sample of blood. For the thin
blood smear, a drop of blood was placed at the end of the glass slide and then spread
across a large area of the slide. Thin blood smears will helps the doctors to discover what
species of malaria is causing the infection in the body. Microscopic examination of thick and
thin blood smears were the easiest and most reliable test for malaria detection.

Last but not least, the prepared slide was stained by using Giemsa staining. Giemsa
staining was used in this experiment to differentiate the nuclear and cytoplasmic morphology
of platelets, RBCs, WBCs, and parasites. Giemsa stain must be diluted for use with water
buffered to pH 6.8 or 7.0-7.2, it is depending on the specific technique used.
 Practical 5a: Preparation of thin and thick blood smear

OBJECTIVE
To prepare thick and thin blood smear for observation of parasite infection

MATERIALS AND METHODS:

A. Preparation of thick blood smear

1. Three small drops of blood was placed onto one end of a clean microscope slide.
2. Using corner of a second clean microscope slide as a stirrer, the contents of the
three drops of blood was combined by thoroughly mixing and spreading to a circular
film approximately the size of a dime (10cent coin) or nickel.
3. The slide was let for air-dry.
4. The hemoglobin was removed by immersing the slide in a buffer solution before
staining or directly during Giemsa staining.

B. Preparation of thin blood smear

1. A small drop of blood was placed close to the end of a microscope slide
2. A second slide (spreader slide) was held on edge at a 30 to 40 degree angle and
draw back into the blood drop, allowing it to spread along the edge of the spreader
slide.
3. Quickly and steadily the spreader slide was pushed forward so that the blood
spreads out into a thin film with a feathered edge.
4. Air-dry and stained.
 Practical 5b: Giemsa staining of thin and thick blood smear

OBJECTIVE
To stain thick and thin blood smear using Giemsa stain.

MATERIALS AND METHOD:

A) Thick smear
1. Thick smear was stained with Giemsa stain (1:50 dilution of Giemsa stock solution
with buffered water for 30-60 minutes)
2. Thick smear was washed briefly under slow running tap water for 3-5 minutes and
drained.
3. Air dries in a vertical position.
4. Then, thick smear was examined under the microsocope
a) Using 10X objective lens, the entire smear for blood helminth was scanned.
b) Using 100X objective lens, malaria parasites was screened.
5. The result was reported.

B) Thin blood smear

1. The prepared slide was immersed for 1 minute in a Coplin jar that contains absolute
methyl alcohol.
2. Let air dry.
3. The prepared slide was stained with Giemsa stain (1:20 dilution) for 20 minutes.
4. The prepared slide was washed by dipping slides in and out of buffered water once
or twice.
5. The prepared slide was let for air-dry in a vertical position and was examined under
oil.
6. The prepared slide was examined under the microscope
a) Using 10X objective lens, the entire smear for blood helminth was scanned.
b) Using 100X obejctive lens, malaria parasited was screened.
7. The result was reported.
RESULTS

Blood smears Image under 100x objective lens

White
blood cell
Thick

Thin Head

White
blood cell

Body

White
blood cell
Tail

White
blood cell
DISCUSSION

Thick and thin blood smear was used to determine wether we have malaria or not.
There arre four species of malaria which commonly infect man and that is Plasmodium
falciparum, Plasmodium vivax, Plasmodium malariae and Plasmodium ovale. The most
important of these malaria is Plasmodium falciparum because it can be rapidly fatal. So, a
way to determine wether someone got malaria or not is by using thin and thick blood smear.

To determine the presence of plasmodium, we used blood as the sample. This is

because cysts and trophozoite live in red blood cell (RBC). We can determine wether
someone got infection or not by observe the shape of RBC, the presence of rings, number of
merozoites and so on. Hence, after done the smear, the best part on the smear to observe
the malaria is at feathered part of edge.This is because at the end of the smear, the cells of
blood will separate properly so it is easy for us to observe it. Normally at te hospital they
used a spreader that have a round edge.

Based on the result above, there is no any presence of plasmodium parasites. It

means that the blood sample from my friend was free from any infection of malaria.There
were also no presence of trophozites and cysts discovered from the blood specimen.

There were a few errors that occur during this experiments. Firstly,the usage of
alcohol in the laboratory. Many students let the alcohol be open directly to the environment
and this can cause dangerous to us. As the prevention, alcohol should be stored in the lock
cupboard and we also should not inhale the alcohol directly. Next, the error that occur is the
blood smear too large or too small. This happened because of the spreader slide was
pushed across the slide in a jerky manner. To get the perfect smear, the spreader slide
should be 45oC before spreading the blood. Lastly, the failure to push the spreader slide
completely across the slide should be avoided so that we can get the perfect blood smear.

There were also a few differences between thick and thin blood smear. Firstly, thick
blood smear has a major disadvantage which is the lysis of RBC’s during the staining
process and it make the slide more difficult to read with the absence of RBC features and
irregularities in the thickness of the film.For thin blood smear, less sensitive than a thick fil
especially where there is a low parasitaemia. This may delay diagnosis if after a prolonged
search no parasites are found. Next, the thick blood film provides enhanced sensitivity of the
blood film technique and is much better than the thin film for detection of low levels of
parasitaemia. Thin blood film is often preferred for routine estimation of the parasitaemia
because the organisms are easier to see and count.

Some precautions step should be taken during this experiments which is once we
have put a drop of blood on the glass slide, do not take too long time to do the smear
because the blood can become clotting so that our smear will not perfect. Next, gloves
should be worn all the times to avoid any contamination to our hands. After that, the
spreader slide should only be used once to prevent carry over of patient’s blood to other
blood smear. Your fingers should be placed on the spreader slide as far down as possible
and apply moderate pressure on the spreader slide. Too high up fingers on the spreader
slide will cause excess pressure that can break and resulting a cut.

CONCLUSION

As the conclusion, I can conclude that by doing blood smears, we can detect
parasites that live in bloodstream by using finger prick technique to get the blood sample for
the identification of parasite infection. The thick and thin blood smears were stained by
Giemsa stain to allow for the detection of white blood cell, red blood cell, and platelet
abnormalities. For this experiment, there was no parasite detected in the blood smears and
this shows that the person is healthy from malarial or microfilariae infections.

REFERENCES
 Lecture Note Clinical Parasitology Csi153 Prepared By Madam Nurhidayah Bt Ab.
Rahim.

 Laboratory Manual CSI 153 Clinical Parasitology, Pn. Nurhidayah binti Ab Rahim.

 http://www.webmd.com/a-to-z-guides/thick-and-thin-blood-smears-for-malaria

 http://www.med-chem.com/pages/lab_procedures/pdf/giemsa_blood_stain.pdf