Serial No. : - 02 Date: - 21.01.

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PREPARATION OF PERIPIIERAL BLOOD SMEAR BY USING WEDGE’S

METHOD

 Aim of the experiment: - To prepare blood film after collecting peripheral blood.

 Principle: - A blood film or peripheral blood smear is a thin layer of blood smeared on a glass
microscope slide and then stained in such a way as to allow the various blood cells to be examined
microscopically.

Theory:

 Definition of RBC (Red Blood Cell) : A type of blood cell that is made in the bone marrow and
found in the blood. Red blood cells contain a protein called hemoglobin, which carries oxygen from
the lungs to all parts of the body. Checking the number of red blood cells in the blood is usually
part of a complete blood cell (CBC) test. It may be used to look for conditions such as anemia,
dehydration, malnutrition, and leukemia. Also called erythrocyte and RBC.

 Definition of WBC (White Blood Cell): A type of blood cell that is made in the bone marrow and
found in the blood and lymph tissue. White blood cells are part of the body’s immune system. They
help the body fight infection and other diseases. Types of white blood cells are granulocytes
(neutrophils, eosinophils, and basophils), monocytes, and lymphocytes (T cells and B cells).
Checking the number of white blood cells in the blood is usually part of a complete blood cell
(CBC) test. It may be used to look for conditions such as infection, inflammation, allergies, and
leukemia. Also called leukocyte and WBC.

 Definition of Platelet: A tiny, disc-shaped piece of cell that is found in the blood and spleen.
Platelets are pieces of very large cells in the bone marrow called megakaryocytes. They help form
blood clots to slow or stop bleeding and to help wounds heal. Having too many or too few platelets

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 or having platelets that don’t work as they can cause problems. Checking the number of platelets in
the blood may help diagnose certain diseases or conditions. Also called thrombocyte.

Fig1.1: Picture of Peripheral Blood Smear

 Requirement: - I) Glass slide 2) Spreader 3) equipment for sterilized finger puncture.

 Procedure:-

a) Take four clean and grease free glass slide and select one as a spreader.

b) Collect the peripheral blood by needle to the tip of middle linger.

c) Discard the first drop of blood.

d) Place a drop of blood on slide in the middle about I cm from the end by touching the slide to
the top of blood drop.

e) Place a smooth clean edge of the spreader slide on the specimen slide just in front of the
blood drop. They should maintain an approximate angle of 30- 45°.

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 f) Using the right hand, draw the spreader back until it touches the drop of blood. Let the blood
run along the edge of spreader.

g) When the blood has spread evenly across the edge of spreader slide, push the Spreader to the
other end of the slide with a smooth quick and controlled movement.

h) Dry the blood smear quickly by waving the slide in the air. If smear does not dry quickly, the
blood cell will shrink.

 Observation and Identification: Various types of leucocytes or WBC are observed under
10x, 45x and 100x magnification under a compound microscope. Erythrocytes and leucocytes
are also being observed.

 Significance: The blood film is made to identify the different types of WBC, RBC and
Platelets.

Diagram of Peripheral Blood Smear

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Fig1.2: Drawing of Peripheral Blood Smear