Dubai Medical College Histology Department Prof. Dr. Nadia M.

El Rouby

BLOOD
Blood is a viscid fluid, formed of plasma fluid (55%) and blood
cells (45%). The plasma is formed of water, organic substances, inorganic
substances, gases, hormones, and enzymes.
Types of Blood Cells and Their Average Number: -
* The number of red blood corpuscles (Erythrocytes) ranges from 4.5 to
5.5 million per cubic millimeter.
*The number of white blood cells or leucocytes ranges from 4000 to
11000 per cubic millimeter.
* The number of Blood platelets or Thrombocytes ranges from 200,000
to 400,000 (average: 1/4 x106) per cubic millimeter.
The Major Functions of Blood: -
1-Transport of oxygen, nutritive substances, hormones, enzymes etc
2- Removal of carbon dioxide and waste products through the lungs,
kidneys and sweat glands.
3- Control of body temperature
4- Maintenance of acid-base balance
5-Protect the body against infections.
WHITE BLOOD CELLS (Leucocytes)
They are colorless, but when they are packed together, they appear white
in color.
General characteristics of leucocytes:
▪ They are true nucleated cells.
▪ They contain both cell organelles and inclusions.
▪ They have an amoeboid movement, so they can penetrate through
the capillary walls to perform their phagocytic function in the
connective tissue outside the blood stream.
▪ They partially resist the changes in osmotic pressure.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

Number of Leucocytes:
Their number varies from 4000 to 11.ooo /mm3 (total leucocytic
count). At birth, the total count is about 16.000 /mm3. The number of
leukocytes in the blood varies according to age, gender, and physiologic
conditions.
Leukocytosis:
❖ The increase in number of leucocytes above normal (12000/mm3)
is called leukocytosis.
Leukocytosis may be:
*Physiological: Occurs in pregnancy, during delivery, in newly born
infants, after cold bath, after heavy meal, after exposure to sun. This is
due to rush of leucocytes from blood capillaries of the internal organs to
the peripheral circulation.
*Pathological: Occurs in acute and chronic diseases and certain fevers.
❖ Leucopenia:
The decrease in number of leucocytes below normal is called
leucopenia. It occurs in typhoid fever, influenza, and bone marrow
affection (after exposure to X-ray or after taking certain antibiotics for
long duration or invasion of bone marrow with malignant cells).
Types of Leucocytes
Five types of leukocytes are normally present in the circulation.
Leucocytes are divided into two main groups based on the nuclear shape
and presence or absence of specific cytoplasmic granules (by using light
microscopy)

A) Granulocytes
1- Neutrophils
2- Eosinophils
3- Basophils
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

B) Non-Granular (agranular) Leukocytes

1- Lymphocytes
2- Monocytes
Granulocytes
They have prominent specific cytoplasmic granules that are
visible by light microscopy. There is also another type of granules
(nonspecific) found. Specific granules are specific for each subtype of
leucocytes and have specific functions. The nonspecific granules present
in all types of leucocytes and called azurophilic granules. These
azurophilic granules are lysosomes.
*Leukocytes leave the capillaries by passing in between endothelial cells
and to the connective tissue by a process called diapedesis.
*Leukocytes normally present in the connective tissue.
❖ Diapedesis: means the unidirectional flow of the granulocytes and
monocytes from the blood circulation to the connective tissues. It is
a constant process. It increased in case of infections. Infection
means entrance of micro-organisms into the body.
1- Neutrophils

Count: 60-70 % of the total leukocytes (TLC).

Dimensions: 10-12 µ in diameter.
Nucleus: Neutrophils have a highly lobulated single nucleus, which is
the most prominent feature. The number of lobes varies from 2-5 lobes

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

connected to each other by fine chromatin. The neutrophils are called

polymorph nuclear leucocytes (PMNL). The number of nuclear lobes
depends on the maturity of the cell. In females, the inactive X-
chromosome (Barr body) appears as drumstick-like appendage attached
to one of the lobes of the nucleus. The mature neutrophil has few
appropriate organelles for protein synthesis, so it has a very limited
capacity to regenerate their lysosomal and specific enzymes which are
rapidly depleted by phagocytic activity. Hence, the neutrophil is
incapable of continuous function, and it degenerates after a single burst
of activity. The life span is four days. The dead neutrophils are called
pus cells.
Neutrophils contain glycogen that helps them to have the ability to
survive in anaerobic environment. This is highly advantageous property
because they can kill bacteria and help clean up debris in poorly
oxygenated areas e.g., in necrotic or inflamed tissues. Neutrophils have
both specific and nonspecific granules. The specific granules contain
alkaline phosphatase, collagenase and lactoferrin enzymes. The
nonspecific granules are lysosomes that have hydrolytic enzymes help in
intra-cytoplasmic digestion of foreign substances.
Functions
❖ Neutrophils are highly motile cells. They can move through the
extra- vascular spaces in a crawling fashion with the help of their
pseudopodia. They are highly phagocytic (they are named
microphages) and their principal function is for acute (bacterial)
inflammation. They ingest and destroy damaged tissues and
invading micro-organisms. Neutrophils attracted to sites of injury
and infection. First, they adhere to vessel wall in a process known
as Margination. This is done by chemotactic factors (cytokines)
that are released from the necrotic tissue. They then migrate
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

(Migration) into the surrounding connective tissue and engulf

bacteria by phagocytosis. Specific granules fuse with the
phagosomes (containing the bacteria) and release their contents to
lower its pH to create a suitable environment for lysosomal
enzymatic action. Then, the azurophilic granules (lysosomes)
release their contents that kill both bacteria and the neutrophil
itself.
❖ Neutrophils secrete trephon substances which help in wound
healing.
❖ Neutrophils secrete pyrogenic substances to elevate the body
temperature and make the environment unsuitable for the growth of
the organism (bacteria). Pyrexia lets the patient to seek the
medical advice.
❖ If the micro-organism is vigorous more neutrophils called to the
site of infection.
❖ If neutrophils fail to deal with the micro-organism, macrophages
(monocytes) attracted to the site of infection.
Clinical application:
1- Neutrophilia:
Increased number of neutrophils above 75% of TLC is called
neutrophilia. This occurs in acute infections as tonsillitis,
appendicitis, and abscess formation.
2- Neutropenia:
The decrease in the number of neutrophils below 50% of TLC
is called neutropenia. It occurs in typhoid fever, T.B., influenza,
severe poisoning, and bone marrow affection (what are the
causes?).
Note:
Several neutrophils’ hereditary dysfunctions have been known.
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

▪ One shows sluggish neutrophils because actin is not polymerized.

▪ In another dysfunction neutrophil fail to produce hydrogen
peroxide (H2O2) and superoxide (H2O3) so their microbial killing
power decreases.
Notice that:
▪ All neutrophils’ dysfunctions are associated with persistent
bacterial infections.

Hypersegmented neutrophils (multilobed polymorphonuclear leukocytes)

are larger than normal neutrophils with five or more segmented nuclear lobes.
They are commonly seen with folic acid or vitamin B12 deficiency.

▪ To differentiate between typhoid fever and appendicitis (each of

them presents with acute abdominal pain): we can do the total
leucocytic count and the differential count. If the total leucocytic
count is above 11.000/mm3 (i.e., leukocytosis) with neutrophils
more than 75% (i.e., neutrophilia), it means appendicitis. If the
total count is less than 4000/mm3 with (i.e., leucopenia) and
neutrophils less than 50% (i.e., neutropenia), it means typhoid
fever.
Differential leucocytic count: it means the percentage of each subtype of
leucocytes in relation to the total leucocytic count.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

2- Eosinophils

These are less numerous about 2-4 % of TLC in peripheral blood.

Dimensions: 14-17 µ in diameter.
They are produced in the bone marrow, and they remain in the bone
marrow for several days after production to become mature and then they
circulate around for 3-8 hours before going to the connective tissue and
stay there.
The nucleus is characteristically bilobed. The two lobes are connected by
thick chromatin.
The eosinophil is slightly larger than the neutrophil and can be
recognized by its large specific granules that stain bright red with eosin.
The most characteristic feature is the presence of large ovoid (elongated)
specific granule, which contains an elongated crystalloid center (seen
by EM). This crystalloid center contains extremely alkaline protein called
major basic protein and other basic proteins. The major basic protein is
used in killing parasitic worms. Lysosomes contain hydrolytic enzymes
as (histaminases, sulphatase and acid phosphatase).
Functions:
❖ Eosinophils are phagocytic cells, and their metabolism is like that
of neutrophils, but they have more oxidative capacity.
❖ Eosinophils are not as microbicidal as neutrophils, but they are
phagocytic for antigen-antibody complex of allergy.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

❖ Eosinophils have receptors for IgE, which is required in the

destruction of parasites. The immunoglobulin and complement
receptors present on the plasma membrane of the eosinophils are
different from those found on the surface of other leukocytes.
Eosinophilia:
Increased number of eosinophils above normal is called
eosinophilia. It is associated with allergic reactions and parasitic
infections.
Eosinopenia:
Decreased number of eosinophils is called eosinopenia. It occurs
after prolonged cortisone treatment because it interferes with the release
of eosinophils from the bone marrow to the blood stream.
3- Basophils

Count is less than 1% of T L C (0 %- 0.5 %).

Diameter: 12-14 um in diameter. This is an intermediate size between
neutrophils (10-12µ) and eosinophils (14-17µ).
The life span: is unknown (why?).
The nucleus: is bi-lobed, irregular, or S- shaped nucleus and usually is
masked by large basophilic granules.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

The cytoplasm: contains numerous large, densely basophilic (deep blue)

specific granules. These granules contain heparin and histamine. The cell
also, contains few free ribosomes, mitochondria, and glycogen.
The plasma membrane: have blunted, irregularly spaced surface
projections and has membrane receptors highly specific for Fc segment of
Ig E, this is an immunoglobulin which is released in response to
allergens. Hence basophils may supplement the functions of mast cells in
immediate hypersensitivity reactions by migrating to connective tissues.
Function:
• They are not phagocytic like eosinophils or neutrophils.
• They secrete heparin and histamine.
Agranulocytes
This group of cells contains cytoplasmic nonspecific granules that
are not easily identified by early microscopic methods.
1- Monocytes

Count: 2-10 % of TLC.

Dimension: up to 20 µm in diameter so monocytes are the largest white
blood cells (macrophages).
The nucleus of monocyte is large, eccentric, and pale. The chromatin is
less condensed and has a more fibrillar arrangement of chromatin than

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

that in lymphocytes. Because of this delicate chromatin distribution, the

nuclei of monocytes stain lighter than do those of large lymphocytes. The
nucleus is either oval, horseshoe- shaped or kidney-shaped. It has two or
more nucleoli.
The cytoplasm is non granular and basophilic. It contains very fine
azurophilic granules (lysosomes) that are distributed throughout the
cytoplasm as well as phagocytosed particles. Also, it has rER,
mitochondria, Golgi, and free ribosomes.
Function:
Monocytes are highly motile phagocytic cells. They are the
precursors of macrophages in the C.T (tissue histeocytes). Monocytes
are capable of continuous lysosomal activity and regeneration of
lysosomal enzymes. Monocytes use aerobic and anaerobic metabolic
pathways depending on the availability of oxygen in the tissue.
Monocytes appear to have little function in the circulating blood. They
usually migrate to the C.T. where they become macrophages. They
respond to the presence of necrotic material, invading microorganisms
and inflammation (these conditions release chemotactic substances) by
migration into the tissue and differentiation into macrophages; with their
great capacity for phagocytosis and large content of hydrolytic enzymes,
the macrophages engulf and destroy tissue debris and foreign material as
part of the process of healing and restoration of normal function.
Life Span: It is about 3 days in the blood stream. They mainly survive in
the C.T.
Monocytosis means an increase in the number of monocytes in the
peripheral blood above 10% of the TLC. This occurs in chronic
infections as Malaria, T.B., and Glandular Fever.
Monocytopenia means decrease number of monocytes is mostly due to
bone marrow causes (what are?) and infection with HIV.
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

2- Lymphocytes
The percentage of lymphocytes in the blood is 20-30 % of T L C.
*lymphocytes are classified into Small and Large lymphocytes.
A- Small Lymphocytes
The percentage: 15-20 % of T L C.
Diameter: is about 6-8 µ in diameter.
The cytoplasm: is scanty and contains few azurophilic granules
(lysosomes) and many free ribosomes so it is basophilic.
The nucleus: is rounded, darkly stained and filling nearly the whole cell
leaving a thin ring of homogenous basophilic cytoplasm.
There are two types of small lymphocytes:
1- T-Lymphocytes: consist 75 % of small lymphocytes.
2- B- Lymphocytes: consist 25% of small lymphocytes.
T & B Lymphocytes are covered by microvilli. Their cytoplasm shows
many free ribosomes, few mitochondria, endoplasmic reticulum, a pair of
centrioles and lysosomes.
b- Large Lymphocytes:
The percentage is: 5 to 10% of TLC.
Diameter is: 10 -15 µ.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

The cytoplasm is abundant, and it contains many ribosomes,

mitochondria, and a well-developed Golgi apparatus.
The nucleus is slightly indented. It is darker than the nucleus of
monocyte. They are considered as activated small lymphocytes after
exposure to an antigen.
*Lymphocytosis:
Increased percentage of lymphocytes above 30% of the TLC is called
lymphocytosis.
Causes of Lymphocytosis:
Chronic diseases as Whooping cough, T.B., and Glandular fever.
N.B.: Lymphocytes can pass through the epithelial covering so that they
might appear in the oral cavity and called salivary corpuscles. A moving
lymphocyte has a head and a tail; it has a shape like tennis racquet.
*Classifications of small Lymphocytes
According to the origin and functions, Small lymphocytes are classified
into:
T-Lymphocytes and B-lymphocytes:
T- Lymphocytes
They called T-Lymphocytes because they are Thymus dependent
cells. They require the presence of the thymus gland for development and
maturation.
Clinical application:
Congenital absence of thymus gland causes absence of T- lymphocytes.
T-lymphocytes represent most of the circulating lymphocytes. They are
about 75% of the circulating small lymphocytes.
They have long life span as they can live for years.
T-lymphocytes develop from mother cells in the bone marrow.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

The mother cells migrate from the bone marrow to the thymus where they
proliferate and change into T-lymphoblasts and then T-lymphocytes
(thymocytes).
There are different types of T- lymphocytes:
1. T- Helper cells: help B-Lymphocytes to perform their functions.
2. T-Suppressor cells: suppress certain functions of B & T –
Lymphocytes.
3. T-Killer cells kill bacteria and viruses.
4. T-Lymphokine secreting cells: secrete lymphokines.
5. T-Memory cells: These memory cells can defend the body against
the same antigen if renters the body for the 2nd time.
*Function of T-Lymphocytes
1- T-lymphocytes are responsible for cellular immunity (cell mediated
immune response). If an antigen enters the body, T-lymphocytes cause
its lysis and destruction.
Therefore, cellular immunity means that T-lymphocytes kill bacterial
cells directly (i.e., cell to cell). So, T-lymphocytes act as Killer Cells.
2- The T-lymphocytes secrete Lymphokines that are hormone-like
factors. They include:
*Interferon which inhibits viral replication.
*Colony stimulating factor which stimulates proliferation of bone
marrow cells.
*Mitogenic factor stimulates the processes of lymphocyte proliferation.
*Cytotoxic factor which kill certain bacteria as T.B. The T cytotoxic
cells act against foreign cells by two mechanisms:
They produce proteins called perforins that create holes in the
bacterial cell membrane with consequent cell lysis.
They kill the cell by activating certain genes that cause apoptosis.
The apoptotic cell is rapidly removed by macrophages.
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

Apoptosis means programmed cell death or natural cell death.

3- T- lymphocytes may act as Graft Rejection Cells. After transplantation
of skin grafts or any organ e.g., kidney, T-lymphocytes of the recipient
migrate and settle under the transplanted organ and secrete lymphokines
that cause rejection of this organ.
4- T-Lymphocytes act as T-Memory cells.
5- T-lymphocytes considered as helper cells to B-lymphocytes. They can
help the B-lymphocytes to react against the specific antigens that may
enter the body. T-cells process and present the antigen for B- cells. This
means that T-cells partially digest the antigen for B- cells.
6- T-lymphocytes act as Macrophage Chemotactic Cells. They activate
and stimulate the macrophage to migrate to sites of infection.
Both T helper and T suppressor cells collectively called regulator cells.
B-lymphocytes:
They are termed B-lymphocytes because they develop from the
primitive stem cells of the bone marrow. They complete their
development & maturation over there. They leave the bone marrow to the
circulation as mature cells.
They constitute the minority of circulating small lymphocytes (about
25%).
They have shorter life span (about 3 months).
Functions of B-Lymphocytes:
* B-Lymphocytes are responsible for the development of humoral
immunity; they are changed into plasma cells which secrete antibodies
that circulate in the blood stream hence the name humoral immunity
(Systemic Blood Immunity).
The development of humoral immunity takes place as follows; if the
antigen enters the body, first, it is picked up by T- lymphocytes that act as
helper cells for B-lymphocytes. The antigen is processed and then
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

subsequently delivered from T-lymphocytes to B-lymphocytes. In this

case the B-lymphocytes enlarge and become activated and then
differentiate into plasmablasts then plasma cells. Plasma cells secrete
antibodies that react specifically with the stimulating antigen.
* Some B-lymphocytes act as B-Memory Cells; the majority of the
activated B-lymphocytes differentiate into plasma cells. Some activated
B-lymphocytes remain in the blood as B-Memory cells. The memory
cells can produce secondary immune response when they are exposed
once again to the same antigen. Secondary immune response is a rapid
response.
N.B. natural killer lymphocytes (NK): these cells lack the surface
marker molecules characteristic for B and T cells. In the circulating
blood, 10-15% of lymphocytes are N K cells. They attack virus – infected
cells and cancer cells without previous knowledge (i.e. without
sensitization).
Blood Platelets (Thrombocytes)
The number is 200,000- 400,000/ mm3 of blood with an average of
1/4 million/mm3. Decreased number is called (purpura or
thrompocytopenia in which there is prolonged bleeding time). Blood
platelets are oval small (2- 4 µ) non nucleated basophilic bodies. They
usually found in the form of clumps in blood film. The platelets have a
central granulomere contains glycogen, lysosomes, and mitochondria.
Delta granules contain calcium ions, ADP, ATP and serotonin. Alpha
granules contain fibrinogen and platelet derived growth factor (now it is
called thrombopoietin). A peripheral hyalomere contains microtubules
and microfilaments. The main functions of blood platelets are secretion of
serotonin, platelet aggregation, blood coagulation and clot retraction.
After this, removal of the clot by macrophages will occur.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

*blood does not coagulate normally in hemophilic patients. Hemophilia

A (deficiency of factor VIII) & B (deficiency of factor IX). Any minor
trauma can cause severe hemorrhage in hemophilic patient.
Red Blood Corpuscles
Number: In males: 5 x 106 to 5.5 x 106 / mm3
In females: 4.5 x 106- 5x 106/ mm3
The number is less in females:
1- Menstruation in females causes loss of blood monthly.
2-The male hormone (testosterone) stimulates the bone marrow.
Abnormalities in the number of R B Cs:
* Decrease in the number is known as Anemia or oligocythaemia
* Increase in the number is known as Polycythaemia.
Anemia: It is a decrease in the number of R B Cs and /or hemoglobin
content below normal (4 million per cubic millimeter).
Oligocythemia: means decreased number of RBCs only.
Causes and Types of Anemia:
1- Deficiency anemia: it is due to deficiency of iron, copper, proteins,
hormones (e.g. erythropoietin), vitamin C, vitamin B12 & folic acid.
2- Hemorrhagic anemia: as hemorrhage from nose, gums & piles.
2- Hemolytic anemia: it is due to excessive destruction of R B Cs as
in Spherocytosis (membrane abnormality) and sickle cell

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

(hemoglobin abnormality). In sickle cell disease, erythrocytes are

inflexible and fragile. They have short life span. There is increased
viscosity of the blood so the blood flow in capillaries decreases
causing anoxia (decreased oxygen level in blood).

Sickle cells. Homozygous sickle cell disease. A nucleated red cell and neutrophil are also in the field.

4- Aplastic anemia is due to bone marrow suppression as in exposure to

irradiation, prolonged use of certain antibiotics or infiltration by
malignant cells as in leukemia (cancer of leucocytes).
*Polycythamia (erythrocytosis): it is an increase in the number of RBCs
above 6 millions /mm3:
* Physiological: (as in hypoxia in high altitudes)
* Pathological: (cases of heart and lung diseases).
*Size of R B Cs: - Normal diameter is 7.5 µ.
Thickness: the normal thickness is 1.9 µ at the periphery and 1.1 µ at the
center of RBCs. This means that the corpuscle is thicker at the periphery
than in the center so more Hb present at the periphery.
Abnormalities in the Diameter of R B Cs:
Macrocytic there is an increase in the diameter of R B Cs above 9 µ.
* Microcytic there is a decrease in the diameter of RB Cs below 6 µ.
*In case of Anisocytosis, R B Cs have different diameters.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

*Shape of R B Cs
They are rounded, biconcave, non-nucleated discs. In slow blood
stream and in blood films, R B Cs adhere together due to surface tension
showing rouleaux appearance.

Acanthocytosis. Spiculated red cells are of two types: acanthocytes are contracted dense cells with
irregular membrane projections that vary in length and width; echinocytes have small, uniform, and
evenly spaced membrane projections. Acanthocytes are present in severe liver disease, in patients with
abetalipoproteinemia, and in rare patients with McLeod blood group. Echinocytes are found in patients
with severe uremia, in glycolytic red cell enzyme defects, and in microangiopathic hemolytic anemia.

*Abnormal Shapes of R B Cs: -

In certain anemias, R B Cs appear biconvex (spherocytes) or oval
(ovalocytes).

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

*Colour of R B Cs
R B Cs are greenish yellow in colour due to presence of
Hemoglobin. A drop of blood appears red due to overlapping of RBCs
that have oxyhaemoglobin. When the hemoglobin % is normal, the red
blood corpuscles are called normochromic.
*Abnormal Colour of R B Cs:
R B Cs with less hemoglobin % than normal are pale and are called
hypochromic.
RBCs with more hemoglobin % than normal are called hyperchromic.
In target cell anemia, hemoglobin is concentrated in the center forming
central colored mass and peripheral pale ring like the target.

Target cells. Target cells are recognized by the bull’s-eye appearance of the cell. Small numbers of
target cells are seen with liver disease and thalassemia. Larger numbers are typical of hemoglobin C
disease.

Contents of R B Cs
They are not true cells; they have neither nuclei nor organelles.
It is surrounded by plastic cell membrane that is lipoprotein in nature.
The RBCs have hemoglobin. Hemoglobin combines with oxygen to form
oxyhemoglobin.
RBCs in blood smears appear as rounded bright, pink-stained cells. The
bright pink colour (acidophilia) is due to their content of oxyhemoglobin
that binds the acidic dye (eosin). Despite lack of organelles, RBCs are
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

metabolically active and derive their energy by anaerobic respiration of

glucose and through ATP generation by the hexose mono phosphate
shunt. Functionally, RBCs are highly deformable and able to be squeezed
through narrow blood capillaries down to 3 µs in diameter. The cell
membrane is braced by cytoskeleton that contains ankyrin spectrin
meshwork. This meshwork is largely responsible for maintaining the
biconcave shape.
RBCs have limited life span and are then destroyed in the spleen.
Their life span is from 100-120 days in the circulation. The senile RBCs
are unable to synthesize new enzymes to replace those lost during normal
metabolic processes. Diminishing efficiency of ion pumping mechanisms
is probably the main factor in RBCs’ ageing. The corpuscle becomes
progressively less deformable until it is unable to pass through the splenic
microcirculation. So, they become easily phagocytosed there.
HEREDITARY SPHEROCYTOSIS
Hereditary spherocytosis is due to abnormal arrangement of the
internal cytoskeleton of RBCs. Normally the internal surface of the cell
membrane is braced by proteins of cytoskeleton via interactions between
ankyrin and spectrin. In hereditary spherocytosis, the ankyrin binding of
spectrin is absent. As a result, the cell membrane is easily deformed.
In hereditary spherocytosis, RBCs do not have biconcave shape, but they
appear spherical and biconvex. They are abnormally fragile and do not
resist changes in osmotic pressure. This causes rapid breakdown of RBCs
(hemolysis). Surgical removal of the spleen improves symptoms of
hereditary spherocytosis due to removal of a large portion of the body's
macrophages that are found in the spleen.
Hemolysis:
Rupture of the cell membrane of RBCs and loss of hemoglobin
outside is known as hemolysis. It may be caused by acids, alkalis, malaria
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

and bacterial toxins, snake venom, hypotonic solution and incompatible

blood transfusion.
Normally the osmotic pressure of R B Cs is 0.9 % saline. If the R B Cs
are exposed to hypertonic solution (2%) shrinkage will occur, while if
they are exposed to hypotonic solution (e.g. distilled water) swelling, then
rupture and hemolysis will occur. Their hemoglobin can be changed into
bile pigments and haemosiderin granules.
❖ Adaptation of the R B Cs to perform their functions:
The main function of R B Cs: is to enclose hemoglobin, this
hemoglobin is formed of a protein (globin) and an iron containing
pigment (haem). Hemoglobin combines with oxygen. Hemoglobin also
plays a role in controlling hydrogen ion concentration of blood.
Hemoglobin can only do these functions when it presents inside the red
blood corpuscles.
The cell membrane of R B Cs is plastic, so the R B Cs are flexible
easily deformable and not rigid. It allows R B Cs to change their
shape and size to be squeezed through narrow capillaries and
regain their normal shape and size in wider vessels.
Because erythrocytes are flexible, the viscosity of the blood
normally remains low.
The biconcave shape of R B Cs increases the surface area for gas
exchange.
The cell membrane of R B Cs is lipoprotein in nature, so it is
highly selective; it allows easy exchange of carbon dioxide and
oxygen through it.
There are neither nuclei nor cell organelles in R B Cs. This
prevents R B Cs from reproduction and leave free space for
hemoglobin.

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 Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

R B Cs are rich in carbonic anhydrase enzyme that plays an

important role in transportation of carbon dioxide from the tissues
to lungs.
The thickness at the periphery (1.9µ) is more than at the center (1.1
µ) so more haemoglobin is found at the periphery close to the cell
membrane.

.
Mononuclear Phagocytic System
These cells form a system of cells that are distributed throughout
the body. Each cell has single nucleus. All these cells have phagocytic
function. They can be identified by vital stain as trypan blue or black
India ink. In vital stain, we inject the living animal with the warm stain
that circulates with the blood to reach all the tissues of the body. The
phagocytic cells of the various tissues pick up trypan blue from blood and
stained with it. After sometime the animal is killed and the organs are
then taken out and processed in paraffin technique. The stained cells can
be demonstrated under the microscope.
Distribution of mononuclear phagocytic cells:
a. Bone marrow: macrophage, histiocytes and Littoral cells.
b. Brain: microglial cells
c. Blood : monocytes
d. C.T.: histiocytes and tissue macrophages (Free & fixed).
e. Lungs: dust cells which phagocytose dust & carbon particles and
heart failure cells that phagocytose red blood corpuscles in cases of
congestive H.F.
f. Spleen: histiocytes in C.T. and macrophages of pulps.
g. Liver: histiocytes, macrophages and Von Kupffer cells.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

Development of Blood Cells

The process of development of different blood cells is known as
haemocytopoiesis. In adult man, the haemopoietic tissues are two types:
1- Myeloid Tissue (Bone Marrow).
2- Lymphoid tissue (Lymph nodes, spleen, tonsils, and thymus)
Myeloid Tissue (Bone Marrow)
Myeloid Tissue includes yellow and Red Bone Marrow.
Inactive Yellow Bone Marrow: - It present in the cavities of long bones
of an adult. It is formed of; many fat cells, reticular cells and some stem
cells. It acts as reserve areas for haemocytopoiesis. It may change into red
active bone marrow in cases of severe blood loss.
Red Active Bone Marrow: it present in most of the fetal bones. In
adult, red bone marrow is found in the sternum, vertebrae, ribs, the skull,
and clavicle and in the pelvic bones i.e. in flat and irregular bones.
The Structure of the Red Bone Marrow (Myeloid Tissue):
The bone marrow is formed of:
a) Fixed cells b) blood sinusoids c) free cells.
1- The fixed cells include:
a) Reticular cells: They are branched cells with large pale nuclei. These
cells form the background of bone marrow.
b) Osteogenic cells: are the immature bone cells that present in the
endostium which lines the bone marrow cavity.
c) Fat cells which are the largest cells in bone marrow.
d) Endothelial cells that line the blood capillaries and blood sinusoids.
e) Pericyte cells: present around blood capillaries and small blood
vessels. It can differentiate into fibroblasts.
f) Macrophages: around blood sinusoids.
2- Blood Sinusoids: These are wide irregular blood channels lined with
endothelial cells. They are surrounded by macrophages which can
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

introduce their pseudopodia into the blood sinusoids to engulf foreign

bodies from the blood stream.
3- Free Cells: They are the developing blood cells which are the
immature erythrocytes and leucocytes.
N.B: If we count the number of immature blood cells in the bone marrow,
we find that the number of immature leucocytes is five times as many as
the number of immature erythrocytes, this is called myeloid: erythroid
ratio (5: 1). This ratio can be explained by the fact that, the life span of
leucocytes is about four weeks while the life span of erythrocytes is about
four months. The bone marrow forms & stores more leucocytes than
erythrocytes because the leucocytes are destroyed rapidly in the blood
stream.

Hematopoietic Precursor cells

*Pluripotential stem cells: (haematopiotic stem cell): cells capable of
asymmetric division (give rise to all types of blood cells (i.e. red blood
corpuscles, leucocytes and megakaryocytes).

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

* Multipotential Progenitor cells (precursor cells or CFUs): capable of

forming a specific type of blood cells but with wide range (e.g. leucocytes
with its five subtypes)
*Committed cells: capable of forming only 1 type of cells (e.g. CFU-E in
erythropoiesis).
*Mature cells: they undergo structural differentiation to form one cell
type only. They are incapable of division.

Stages of Erythropoiesis
Erythrocytes develop in the red bone marrow through the following
stages:

1- Hematopoietic Stem Cell (Pluripotential stem cells):

It is an embryonic branched cell present in the bone marrow.
It has large pale nucleus.
It can differentiate into:
2- Colony Forming Unit Cell (Multipotential progenitor cells): It is a
rounded cell from 8-10 µs. It can differentiate into: -
3- Colony forming Unit for Erythropoietin (CFU-E):
It is committed progenitor cell.
It is a rounded cell (10-12µ) with slightly basophilic cytoplasm.
Its nucleus is rounded with fine chromatin granules.

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Erythropoietin hormone is secreted from the kidney. It stimulates these

cells to synthesize hemoglobin.
It differentiates into:
4- Proerythroblast:
It is a rounded cell, from 12 -15 µ in diameter. It has large rounded
nucleus occupies 80% of the cell volume with fine chromatin and two
nucleoli. The cytoplasm is basophilic because it contains ribosomes.
- The cell can differentiate into basophilic erythroblast.
5- Basophilic Erythroblast:
- It is about 10-13 µ in diameter.
- The nucleus is small with condensed chromatin.
- The cytoplasm is more basophilic and is rich in ribosomes.
- It can differentiate into:
6- Polychromatophil Erythroblast:
- It is about 8 - 11 µ in diameter.
- Its nucleus is smaller, dense and nucleoli are not seen.
- The cytoplasm is polychromatophilic (i.e. it shows two colors).
- The acidophilia of the cytoplasm is due to starting appearance of Hb.
- The basophilia of cytoplasm is due to presence of ribosomes.
- This cell differentiates:
7- Orthochromatic Erythroblast (normoblast):
It is about 8-10 µs in diameter.
The cytoplasm is acidophilic due to more Hb. (ortho: means red)
The nucleus is small, eccentric and deeply stained (pyknotic).
It divides into smaller cell containing nucleus (phagocytosed by
macrophages) and the larger cell enters the blood sinusoids as
reticulocytes. Some authors mention that the normoblast extrude its
nucleus to give reticulocyte without division.
8- Reticulocytes:
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

- It is a small cell with a diameter about 8 µ.

- The cytoplasm contains ribosomes in the form of a basophilic reticulum.
- It matures into RBC within 24-48 hours.
- Reticulocytes may appear in the circulating blood, but their percentage
is not more than 1%. Their number may increase after hemorrhage, in
hemolytic crisis or recent ascent to high altitude which means more active
bone marrow to compensate so it pushes to the circulation less mature
cells.
They can be stained with supra-vital stain as brilliant cresyl blue. This
can be done by mixing a fresh drop of blood with the dried powder of this
stain. After a short period of time we spread a blood film form the
mixture. From this blood film we can get out the ratio between
erythrocytes and reticulocytes. Reticulocytes appear as pale pink cells
with basophilic reticulum.
*Mature Erythrocytes: - They are biconcave rounded non-nucleated
discs filled with Hb. They enter through the blood sinusoids of the bone
marrow to reach the circulation.

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

Development of Granular Leucocytes

(Neutrophil, Eosinophil and Basophil)
They develop in the red bone marrow through the following stages:
*Hematopoietic stem cells: - They are small embryonic branched cells
with large nuclei and basophilic cytoplasm.
*Colony forming Unit (CFU): - They are small, rounded cells with
small, rounded nuclei. They differentiate into:
*Colony Forming Unit Granulocytes: - (CFU – G): these rounded cells
differentiate into:
*Myeloblast: - It is a large, rounded cell. The cytoplasm is basophilic
and is devoid of granules. It has a very large nucleus with two or more
prominent nucleoli.
*Promyelocyte: - It is a large cell with a large nucleus and prominent
nucleolus. Its cytoplasm contains azurophilic non-specific granules.
*Myelocytes: (Neutrophil, Eosinophil and Basophil Myeloytes): - Its
cytoplasm contains specific granules. The following three types of cells:

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

*Basophil myelocytes with basophilic granules.

*Neurophil myelocytes with neutrophilic granules.
*Eosinophil myelocytes with acidophilic granules.
Myelocytes cannot divide again. After this stage, they undergo further
nuclear maturation to be changed into Metamyelocytes.
*Metamyelocytes: (neutrophil, Eosinophil and Basophil): differentiation
of nuclei occurs.
N.B. The Neutrophil Metamyelocyte is characterized by the presence of a
kidney shaped nucleus and called Juvenile Neutrophil. Continuous
maturation of neutrophil leads to appearance of Band or Stab neutrophil
with a bend-rod nucleus. These cells may appear in the peripheral blood,
but their percentage normally is not more than 2%. If the percentage of
these Juvenile cells exceeds 2%, this is bad sign. It means severe
infection. So, as a trial from the bone marrow to overcome this infection
it pushes immature neutrophils to the circulation.
*Mature granular leucocytes with their characteristic nuclei and their
specific granules

Hematopiotic stem cell

C F U cell

C F U Granulocyte

Myeloblast

Promyelocyte

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

Myelocyte

Metamyelocyte

Mature Granulocytes

Development of Lymphocytes
All lymphocytes originate in the bone marrow however; T-
lymphocytes development and maturation occur in the thymus, but B
lymphocytes leave the bone marrow as mature cells. For this reason, the
bone marrow and thymus are called primary or central lymphoid
organs. Lymphocytes migrate from central lymphoid organs to the blood
and peripheral lymphoid organs (spleen, lymph nodes, solitary nodules,
tonsils, appendix and Peyer’s patches of the ileum) that are also known as
secondary lymphoid organs.

Development of Monocytes
*Hematopoietic stem cells: - They are branched embryonic cells.
*Colony Forming Unit (CFU):- They are small rounded cells with small
rounded nuclei and basophilic cytoplasm, they differentiate into:
*Monoblast: - it is large cell with large non-indented nucleus. It can
differentiate into:
* promonocyte with slightly indented nucleus. This cell differentiates
into:
*Monocyte: - it is large cell with large indented nucleus. Sometimes
migrates to the tissues to be changed into macrophage.
Development of Blood Platelets
*Hematopoietic stem cell: It differentiates into:
*Colony Forming Unit (CFU- MK): it differentiates into:
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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

*Megakaryoblast: It is a large cell with a rounded slightly indented

nucleus which has two clear nucleoli.
The cytoplasm is basophilic as it is rich in ribosomes.
* Promegakaryocyte:-
It is a larger cell with a large multilobed nucleus.
Its cytoplasm is basophilic and contains fine azurophilic granules.
The cell can differentiate into megakaryocyte.
*Megakaryocyte:-
It is a large cell with a large multilobed nucleus. It is (40-100 µ).
With E/M the cytoplasm appears as if it is divided into small areas. These
areas are then separated as buds and form blood platelets. These platelets
enter the blood sinusoids to the general circulation.

Hematopeiotic stem cell

Megakaryocyte

CFU -MK

Megakaryoblast

Promegakaryocyte

platelets

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Dubai Medical College Histology Department Prof. Dr. Nadia M. El Rouby

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