ASIA METROPOLITAN UNIVERSITY

BACHELOR OF BIOMEDICAL SCIENCE

BLOOD SMEAR AND STAINING

LAB REPORT

NAME: SHARMILY A/P PUNITHAN

STUDENT ID: BBMED20220900173

LECTURER’S NAME: DR. KALAIVANI BATUMALAIE

SUBJECT: BASIC HEMATOLOGY

SUBJECT CODE: HBBF1132

Title Blood Smear and Staining

Objective  To prepare thick blood film

 To prepare thin blood film
 To stain using Leishman stain

Introduction A blood smear is a blood test used to look for

abnormalities in blood cells. The three main blood cells that the
test focuses on are red blood cells that carry oxygen throughout
the body, white blood cells that help your body fight infections
and other inflammatory diseases and platelets that is important
for blood clotting.
The test has a wide range of uses. It can be used to
determine whether an infection is viral or bacterial. It can also
detect anemia, determine the cause of jaundice, and diagnose
malaria.2
A blood smear is different from an automated test like a
complete blood count (CBC). A technician or health care
provider usually examines the blood smear under a
microscope. Blood film results and interpretation can provide
clues in diagnosing the problem.

Materials  Blood sample

 Microscopic slides

 Lancets

 Alcohol swab
 Leishman stain
 Methanol
 Distilled Water
 Dropper
 Compound Light Microscope

Procedure Retrieving blood sample:

1. The microscopic slides were cleaned with 70 to 90%
 alcohol and allowed to dry completely. The surface of
slide where the blood smear is done is not touched.
2. A finger is selected to puncture. (Middle or Ring
finger)
3. Clean the area to be punctured with alcohol swab and
allow it to dry.
4. Puncture the ball of the finger.
5. Wipe away the first drop of blood with alcohol swab.
6. Touch the next drop of blood with a clean slide. Repeat
with several slides.

Preparation of Thin blood smear:

1. Bring a clean spreader slide. The slide is held at 45˚
angle toward the drop of the blood on the slide.
2. Wait until the blood spreads along the entire width of
the spreader slide.
3. Push it forward rapidly and smoothly.
4. Fix the thin blood smear with 100% methanol for 5
minutes and let it dry.
5. Add several drops of Leishman stain to the sample and
stain it for 2 minutes.
6. Drain the slide and wash with distilled water. Let it dry
completely.
7. Observe the sample under microscope using 10x power.

Preparation of Thick blood smear:

1. Using the corner of a clean slide, spread the drop of
blood in circular motion.
2. Let it dry. Make sure the blood smear is not too thick.
3. Observe the sample under microscope using 10x power.
Results Thick blood smear:

Thin blood smear:

Head Body Tail

Thick blood smear under microscope:

 10x

Thin blood smear under microscope:

 10x

Discussion The normal red cell is biconcave disc-shaped, measures

about 7–8 µm in diameter, has central pallor (approximately a
third of the red cell diameter) and lacks intra-cytoplasmic
inclusions. Red cells are pink in color when stained with
 Rowmanosky dye because the haemoglobin content of the red
cell picks up eosin, the acidophilic components of the dye.8
Abnormal variations in cell size, shape, colour, presence of
intracellular inclusions and pathologic arrangement of the cells
suggests a host of abnormalities.
Leishman stain is used in this experiment because it is
one of the better stains for routine blood staining of peripheral
blood clotting for blood film under a microscope and is
satisfactory for malaria and other blood parasites. Leishman
stain is a combination of methylene blue and eosin dye,
prepared in an alcohol medium and diluted with buffer or
purified water during the staining process. Leishman stain is a
differential stain that is used to stain the various components of
the cells and can be used to study the adherence of pathogenic
bacteria to human cells. It colours human and bacterial cells
distinctly and acts as purple and pink coloured bodies,
respectively.

Conclusion A blood smear is considered normal when your blood

contains a sufficient number of cells and the cells have a
normal appearance. A blood smear is considered abnormal
when there’s an abnormality in the size, shape, color, or
number of cells in your blood. Abnormal results may vary
depending on the type of blood cell affected.