INTRODUCTION TO BASIC

HISTOLOGY
BY

Dr. (Mrs.) O. A. Ebeye

Ahama E. Efe
BLOOD
 Blood
• Blood is a specialized fluid connective tissue in
which cells are suspended in fluid extracellular
material called plasma.

• Blood cells: erythrocytes, leukocytes and platelets

are found in blood

• Blood tend to clot once its leaves the circulatory

system
• Collected blood in which clotting is prevented by the addition of
anticoagulants (eg, heparin or citrate) can be separated by
centrifugation into layers that reflect its heterogeneity.

• A straw-coloured, translucent, slightly viscous supernatant

comprising 55% at the top half of the centrifugation tube is the
plasma

• A thin gray-white layer called the buffy coat of about 1% of the

volume, consists of leukocytes and platelets, both less dense than
erythrocytes.

• Erythrocytes (hematocrit) makes up the sedimented material and

their volume is normally about 45% of the total blood volume in
healthy adults.
Composition of Blood
 Other features and Function of Blood
• Act as a distributing vehicle, that transports O 2, CO2, metabolites,
hormones, and other substances to cells throughout the body.

• O2 binds mainly to hemoglobin in erythrocytes and is much more

abundant in arterial than venous blood, while CO 2 is carried in
solution as CO2 or HCO3–, in addition to being hemoglobin-bound.

• Nutrients are distributed from their sites of synthesis or gut

absorption, while metabolic residues are excreted from cells all over
the body

• Hormonal distribution in blood permits the exchange of chemical

between varying distant organs, regulating normal organ function.

• Blood also participates in heat distribution, homeostasis, and the

 Plasma Composition
• Aqueous solution, pH of 7.4, containing
substances of low or high molecular weight that
make up 7% of its volume.

• It is composed mostly of water, plasma proteins

and other solute (mostly electrolytes).

• The composition of plasma is usually an indicator

of the mean composition of the extracellular fluids
in tissues.
 Blood Cells
• This is the solid part of blood, containing several
cells.

• It is histologically studied by preparing a smear,

composed of mixture of dyes: eosin and
methylene blue.

• These dye mixture also contain dyes (azures),

which are useful in staining cytoplasmic granules
containing charged proteins and proteoglycans.
Making a Blood Smear
 Erythrocytes (RBCs)
• There are terminally differentiated structures lacking nuclei and
completely filled with the O2 carrying protein hemoglobin.

• Only blood cells whose function does not require them to exit
circulation, and adhere to one another (Rouleaux).

• They are 7.5 μm in diameter, 2.6 μm thick at the rim and 0.75 μm thick
in the center, which is used by histopatologist as a standard to
estimate the size of other cells.

• The biconcave shape provides a large surface to volume ratio and

facilitates gas exchange.

• The normal concentration of erythrocytes in blood is approximately

3.9 - 5.5 million per microliter (μL, or mm 3) in women and 4.1 - 6.0
million/μL in men.
• The erythrocyte is composed of a membrane known as the
plasmalemma, composed of 40% lipid, 10% carbohydrate and 50%
protein.

• About half the protein span the lipid bilayer and is composed of integral
membrane protein, ion channels and anion transporter.

• The glycosylated extracellular domains of the latter proteins of

plasmalemma include antigenic sites that form the basis for the ABO
blood grouping system.

• Erythrocyte cytoplasm lacks all organelles but is densely filled with

hemoglobin and tetrameric O2 carrying protein.

• N/B: Erythrocyte differentiation includes loss of the nucleus and

organelles, shortly before the cells are released by bone marrow into
the circulation
Diagram of an Erythrocyte
 Leukocytes (WBCs)
• They leave the blood and migrate to the tissues where they become
functional and perform various activities related to immunity and
defense.

• They are divided into granulocytes and agranulocytes and are spherical
while suspended in blood plasma, amoeboid and motile after leaving the
blood vessels to invade tissues.

• Granulocytes possess two major types of cytoplasmic granules:

lysosomes and specific granules. They have polymorphic nuclei with two
or more distinct nuclear lobe.

• Agranulocytes do not have specific granules, but contain azurophilic

granules (lysosomes), with affinity for the basic stain. The nucleus is
spherical or indented but not lobulated.
Types and Sub Classification
How Leukocyte Basically Function (Chemotaxis)
 Neutrophils (Polymorphonuclear Leukocytes)
• Mature neutrophils are 54% to 62% and immature ones forms 3% to 5% of
circulating leukocytes.

• Neutrophils are 12-15 μm in diameter in blood smears, with nuclei having

two to five lobes linked by thin nuclear extensions.

• Neutrophils are inactive and spherical while circulating but become actively
amoeboid during diapedesis and upon adhering to solid substrates such as
collagen in the ECM.

• They are usually the first leukocytes to arrive the sites of infection, where
they actively combat bacterial cells through chemotaxis.

• Both cytoplasmic granules are present in neutrophil and with great

resemblance appearing as large, dense vesicles and playing a role of
engulfing microorganisms. They contain proteases and antibacterial
proteins, including the following: myeloperoxidase, lysosome and defensins.
Micrograph of a Neutrophil
 Eosinophils
• Eosinophils are far less numerous than neutrophils, constituting only 1% to
3% of leukocytes.

• In blood smears, their cells is about the same size as a neutrophil or

slightly larger, having bilobed nucleus.

• The main identifying characteristic is the abundance of large, acidophilic

specific granules typically staining pink or red.

• Ultrastructurally the eosinophilic specific granules appears oval in shape,

containing major basic proteins (MBP) that accounts for the granule’s
acidophilia and constitutes up to 50% of the total granule protein.

• Eosinophils are abundant in connective tissue of the intestinal lining and at

sites of chronic inflammation.
Micrograph of a Eosinophils
 Basophils
• They composed of less than 1% of blood leukocytes and are difficult
to find in smears of blood sample.

• They are about 12 - 15um in diameters. Their nucleus is divided into

two irregular lobes, having large specific granules overlying the
nucleus which usually obscure its shape.

• The specific granules (0.5μm in diameter) typically stain purple with

the basic dye (metachromasia) of blood smear stains and are fewer,
larger, and more irregularly shaped than the granules of other
granulocytes.

• This is due to the presences of heparin and histamine, which causes a

similarity between the granules of basophils and mast cells.
Micrograph of a Basophils
 Lymphocytes
• They are composed of spherical cells with similar structural features
and play diverse functions with respect to defend.

• They are small sized which are about 6 - 8um in diameters, medium and
large sized with diameter which are up to 18um.

• The small lymphocyte are predominant in blood having a condensed

nucleus, with a scanty cytoplasm and appears as a thin rim on blood
smears.

• They are slightly basophilic, assuming light blue color in stained smears.
Their life span vary, as some lived only a few days and others survive in
the circulating blood for many years

• They are the only leukocytes that return from the tissues back to the
blood after diapedesis.
Micrograph of a Lymphocytes
 Monocytes
• They are bone marrow derived agranulocytes with varying diameter of
12 - 20um. The nucleus is oval, horseshoe or kidney shaped and
generally eccentrically placed.

• They possess precursor cells of macrophages, osteoclasts, microglia,

and other cells of the mononuclear phagocyte system in connective
tissue.

• They also posses a dense chromatin than that of lymphocytes and

cytoplasm is basophilic, containing azurophilic granules, making the
granule to appear as bluish gray colour in stained smear.

• They also possess 2 nucleoli in their nucleus, RER, polyribosomes, golgi

apparatus and small mitochondria.
Micrograph of a Monocytes
 Platelets (Thrombocytes)
• They are non nucleated disc like cell fragment of 2 - 4um in diameter,
originating from polyploid megakaryocytes located in BM.

• They appear in clumps and as a peripheral light blue transparent

(hyalomere) and a central purple granule (granulomere) zone in blood
smears.

• The hylomere possess actin and myosin molecule, that aids contractility
and movement of platelets

• The granulomere possess bound granules, with sparse population of

mitochondria and glycogen particles. They appear in 2 forms: gamma and
alpha granules

• They contain platelet-derived growth factor (PDGF), platelet factor 4, and

several other platelet-specific proteins.
 How Platelet Controls Hemorrhage
• Primary aggregation – Platelet plug formed.

• Secondary aggregation – Plug releases platelet aggregation

inducers (specific adhesive glycoprotein).

• Blood coagulation – Aggregation of blood (fibrinogen)

component forms a cascade (fibrins) of network of blood
clot.

• Clot retraction – Interaction of actin, myosin and formed ATP.

• Clot removal – Restoration of new formed tissue

Micrograph of a Platelets
 Question
• What is the relation of aspirin as an anti-
inflammatory agents to normal platelet
function
Thank You