Practical 3

Blood Smear Preparation

Blood
Blood is the fluid that circulates in the body and carries substances such as nutrients and oxygen
to the tissues and carbon dioxide and metabolic wastes away from the tissues.

Blood is composed of plasma and blood cells.

Plasma

Plasma is the relatively clear, yellow fluid in which blood cells are suspended. Normally, 55% of
our blood's volume is made up of plasma. As the heart pumps blood to cells throughout the body,
plasma brings nourishment to them and removes the waste products
of metabolism. Plasma contains of 90% water. Plasma also contains blood clotting factors,
sugars, lipids, vitamins, minerals, salts, hormones, enzymes, antibodies, and other proteins.

Blood cells
Blood cells include

Red blood cells (Erythrocytes)

The red blood cells, which are also called erythrocytes (erythro means red; cytes cells) have the
important responsibility of carrying the oxygen throughout the body. Red cells, are relatively large
microscopic cells without nuclei. Mammalian red blood cells are typically shaped as biconcave
disks: flattened and depressed in the center. Red cells normally make up 40-50% of the total blood
volume. The Hemoglobin exists in the red blood cells. Hemoglobin is the gas
transporting protein molecule that makes up 95% of a red cell. One red blood cell contains about
200 million hemoglobin molecules.

White Blood Cells (Leucocytes)

Leucocytes are the cells of the immune system that are involved in protecting the body against
both infectious disease and foreign invaders.
The different types of WBC's are Basophils, Eosinophils, Neutrophils, Monocytes, B- and T-cell
lymphocytes.

Basophils

They are the rarest of the white blood cells (less than 0.5% of the total count) and share
physicochemical properties with other blood cells, they are difficult to study. They can be
recognized by dark violet granules, the nucleus is bi- or tri-lobed, but it is hard to see because of
the number of granules that hide it. Basophils are chiefly responsible for allergic and antigen
response. Basophils can also release chemical signals that attract eosinophils and neutrophils to an
infection site.

Eosinophils

They primarily deal with parasitic infections. They secrete chemicals that destroy these large
parasites, such as hook worms and tape worms that are too big for any one WBC to phagocytize.
In general, their nucleus is bi-lobed. The lobes are connected by a thin strand. The cytoplasm is
full of granules.
Lymphocytes

Lymphocytes are much more common in the lymphatic system than in blood. Lymphocytes are
distinguished by having a deeply staining nucleus that may be eccentric in location, and a
relatively small amount of cytoplasm. Lymphocytes include:

B cells

B cells make antibodies that can bind to pathogen, block pathogen invasion, activate the
complement system, and enhance pathogen destruction.

T cell

The main job of T-cells is to fight infection. There are a number of different types of T-cells that
act in many ways to identify, directly attack and destroy infectious agents. Along with other WBCs,
they play a major role in the immune system, which guards the body against infection.

Neutrophils

Neutrophills are the most abundant white blood cell, constituting 60-70% of the circulating
leukocytes. They defend against bacterial or fungal infection. They are usually first responders to
microbial infection; their activity and death in large numbers forms pus . They have a multi-lobed
nucleus, which consists of three to five lobes connected by slender strands.

Monocytes

These are the largest of all leukocytes. They are amoeboid in shape, having agranulated cytoplasm
Monocytes have unilobar nuclei. Monocytes constitute 2% to 10% of all leukocytes in the human
body. They play multiple roles in immune function. Such as replenishing resident macrophages
under normal states, and (response to inflammation signals, monocytes can move quickly to sites
of infection in the tissues and divide/differentiate into macrophages and dendritic cells to elicit an
immune response.

Platelets
Platelets, also called thrombocytes that play role in blood clotting. Platelets are biconvex discoid
(lens-shaped) structure. They aids in the process of hemostasis ("blood stopping"). Platelets secrete
factors that increase local platelet aggregation (e.g., Thromboxane A), enhance vasoconstriction
(e.g., Serotonin), and promote blood coagulation (e.g., Thromboplastin)

Their production is regulated by the hormone called Thrombopoietin. The circulating life of a
platelet is 8–10 days. The sticky surface of the platelets allows them to accumulate at the site of
broken blood vessels to form a clot. They do this by adhering to the walls of blood vessels, thereby
plugging the rupture in the vascular wall. They also can release coagulating chemicals which
cause clots to form in the blood that can plug up narrowed blood vessels. Thirteen different blood
clotting factors, in addition to platelets, need to interact for clotting to occur. They do so in a
cascading manner, one factor triggering another. Hemophiliacs lack the ability to produce either
blood factor 8 or 9.
PURPOSE OF BLOOD SMEAR
To evaluate red blood cells (RBCs), white blood cells (WBCs), and platelets, to distinguish
between the different types of WBCs, and to determine their relative percentages in the blood; to
help detect, diagnose, and/or monitor a range of deficiencies, diseases, and disorders involving
blood cell production, function, and lifespan

PREPARATION OF BLOOD CELL SLIDE

Put a drop of drop of blood on clean glass slide
Make the smear of blood with the help of other slide
Allow the slide to dry completely
Fix the smear by adding alcohol
Put few drops of methylene blue on slide and mix by shaking the slide
Leave the slide for 5 to 10 minutes
Wash the slide under a gentle stream of water
Leave the slide to dry
Examine the slide under microscope
Human Blood Smear