Al-Qalam University college

Medical Lab. Techniques Dept.

Teaching package

in
Clinical Hematology
Blood cells

Dr.Aydin S.Ahmad

Dr Aydin S A 1
REFERENCE

Dr Aydin S A 2
ATTENTION
Phones should be on silent mode or switched off and
they should be put down on the cupboard or inside
your bag.
After 5 minutes past the start of the lecture, students
are not allowed in the hall
Quiz: You are expected to have a quiz on the present
or previous lecture every week.
Lectures will be uploaded on the group channel a day
before your lecture.
If you have any special inquiries or suggestions please
don’t hesitate to discuss them with me.
 Blood cell Formation

Blood cell formation (haemopoiesis).

The processes that regulate
haemopoiesis and the early stages of
formation of red cells (erythropoiesis),
granulocytes and monocytes
(myelopoiesis) and platelets
(thrombopoiesis)
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 Blood cells

Red Blood cells (erythrocytes) White Blood cells (Leucocytes) Platelets (thrombocytes)

Granular Agranular

Lymphocyte Monocyte
Neutrophils basophils eosinophils

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 Hematopoiesis
 Blood cell formation
 All blood cells come from
pluripotent hematopoietic
stem cells (hemocytoblasts)
– reside in red bone
marrow
– give rise to 5 types of
precursor cells
– precursors develop into
RBCs, WBCs and “giant”
megakaryocytes which
produce platelets by
cytoplasmic
fragmentation
 Sites of haemopoiesis.

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 Overview: Composition of Blood
 A liquid connective tissue
 A mixture
– the formed elements - living blood cells & platelets
– the plasma – the fluid matrix
 Denser and more viscous than water
– due to dissolved ions & organic molecules, especially plasma proteins,
and to the blood cells
– composition and volume regulated by CNS & hormones
 Temp - 38° C
 pH - 7.4 (critical to be between 7.35 and 7.45)
 Volumes differ between sexes, conditional on many factors
– Females - average 4-5 L
– Males - average 5-6 L
9  Salinity : (tonicity) 0.9 % Na Cl (0.85)-
Dr Aydin SA Isotonic
 Overview: Composition of Blood

 Blood sample
– spin it
– separates into 2 parts
• plasma
– ~55% of the volume
– straw colored liquid on top
• formed elements - ~45% of the volume
– red blood cells
– buffy coat - white blood cells and platelets
– Hematocrit = “packed cell volume”PCV
• percentage of formed element measured in a blood sample
• about 45%
Serum= Plasma – (fibrinogen + some clotting factors)
Blood Components
Components of Blood - Plasma
Plasma
–92% water
–7% proteins
–1% other
solutes
Components of Blood - Plasma
 Proteins important for
osmotic balance
– albumin (60%)
• transports lipids
• steroid hormones
– fibrinogen (4%) -
blood clotting
– globulins (35%) –
many different
proteins with a wide
variety of functions
– globulin classes α, β,
and γ
– 1% other regulatory
proteins
Components of Blood - Plasma
 Other solutes
– Waste products -
carried to various
organs for removal
– Nutrients – glucose
and other sugars,
amino acids, lipids,
vitamins and minerals
– Electrolytes (ions)
– Regulatory substances
• enzymes
• hormones
– Gases - O2, CO2, N2
Components of Blood - Formed Elements
 Erythrocytes, or Red
Blood Cells (RBC’s), for
O2 and CO2 transport

 RBCs’ hemoglobin also

helps buffer the blood

IMPORTANT!
Note the
differences in
relative size and
appearance!
RBC Structure

 ~280 million Hgb

molecules/cell

 Hgb for O2 transport

 Bi-Concave shape
greater surface
area/volume ratio increases
gas diffusion
flexibility allows passage
through narrow capillaries
 Red Blood cell(Erythrocytes)
The human Red cell is normally a circular, non
nucleated, biconcave disc.
The red blood cells contain hemoglobin. The
surface area of the red cell is much greater
than that of a sphere of the same size.
Thus the exchange of oxygen and carbon
dioxide is maximal limit with the biconcave
configuration. This shape also helps its to
withstand osmotic lysis and to easily pass
through narrow capillaries.
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Red Cell Dimensions :
Shape: Biconcave disc
Size :7.5 (7-8)µ
Thickness :0.2 mµ
Surface area :140 mµ ²
lifespan of red cells is about 100-120 days.
The dead red cells are broken down by
the reticuloendothelial system

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Formation :
The formation of red cells is known as
erythropoiesis.
During fetal life ,erythropoiesis occure in spleen,
liver, thymus and bone marrow. After birth, it is
confined to the red marrow.
In postnatal life, erythropoiesis takes place in
the bone marrow.
In children, cells are produced in the marrow
cavities of all the bones.

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 Production of Erythrocytes

 Erythropoiesis
– RBC production
– controlled by hormones, especially erythropoietin (EPO) from the
kidney
– three phases of RBC maturation
• production of ribosomes
• synthesis of hemoglobin
• ejection of the nucleus and reduction in organelles
– leave bone marrow as reticulocytes  mature in the blood stream to
become erythrocytes
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RBC Life Span
Functions :
The main function of RBCs is to carry oxygen
from the lungs to the tissues of the body .Red
cells also transport carbon dioxide from the
tissues to the lungs.

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 Normal values :
In adults:
males: (4.5- 6 (million/mm³ of blood
Females: (4 -5.5) million/mm³ of blood
In Newborn: (6-8) million/mm³ of blood

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 Red cell metabolism

 Embden–Meyerhof pathway
 Hexose monophosphate (pentose
phosphate) shunt

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Red cell metabolism
Embden–Meyerhof pathway
In this series of biochemical reactions,
glucose that enters the red cell from plasma
by facilitated transfer is metabolized to
lactate . For each molecule of glucose used,
two molecules of ATP and thus two
high‐energy phosphate bonds are
generated. This ATP provides energy for
maintenance of red cell volume, shape and
flexibility.
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Hexose monophosphate (pentose
phosphate) shunt
Approximately 10% of glycolysis occurs by
this oxidative pathway in which
glucose‐6‐phosphate is converted to G6‐PD
In one of the most common inherited
abnormalities of red cells,
glucose‐6‐phosphate dehydrogenase (G6PD)
deficiency, the red cells are extremely
susceptible to oxidant stress .
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Leukocytes
 Leukocytes (White Blood
Cells) contain nucleus.
– Granular leukocytes
(granulocytes)
• neutrophils
• eosinophils
• basophils
– Agranular leukocytes
(agranulocytes)
• lymphocytes - T
cells, B cells
• monocytes 
tissue
macrophages
 Differential WBC Count

Eosinophil
Lymphocyte 2-4%
20-25%

Monocyte Neutrophil
3-8% 60-70%

Basophil
0.5-1%
Neutrophils
• 62% of total WBCs
• Diameter: 10-12
microns
• Cytoplasm: very
pale blue
• Granules: staining
(purple)
• Nucleus: dark
purple blue 2-5 lobes
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Eosinophil
• Diameter: 13-14
microns
• Cytoplasm : full of
orange-red coarse
granules
• Nucleus: blue 2 lobes
like a pair of glass
• 2-3% of total WBCs
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Basophil
•Diameter: 14-15
•Cytoplasm:
Granules: dark blue
–black obscure
nucleus
•Nucleus: blue
•0.4% of total WBCs
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Lymphocyte
• Diameter: small 7-9
large 12-16
• Cytoplasm : rim, clear,
pale blue
• Nucleus: purple red
,oval indented
• 30% of total WBCs
• Lymphocytes are the
smallest WBC.
• They have large
condensed nucleus, with a
scanty pale blue
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cytoplasm.
Monocyte
• Diameter: 16-18(are the
largest normal blood cells)
• Cytoplasm: The
cytoplasm is abundant, sky
blue in color.
• Some have vacuoles in
the cytoplasm.
• Nucleus: purple, large
kidney shaped and slightly
indented.
•5.8% of total WBCs
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 Lymphocytes - Physiology
Two main types of lymphocytes
– B-cells
• particularly active in attacking bacteria
• develop into plasma cells to produce antibodies (Ab)
– bind to antigen to form antibody-antigen (Ag-Ab) complexes
– complexes prevents Ag from interacting with other body cells or
molecules
– memory B cells – dormant until future exposure to Ag
– T-cells
• attack viruses, fungi, transplants, cancer, some bacteria
• 4 types of cells
– cytotoxic (killer) T cells - destroy foreign invaders
– helper T cells - assist B cells and cytotoxic T cells
– suppressor T cells – help bring immune response to an end
– memory T cells - dormant until future exposure to Ag
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 Leukocyte Life Span
-Life span determined by activity
– Ingesting foreign organisms, toxins, shortens life
– Healthy WBC's – majority last days, but some last
months to years

– During infection, WBCs may only live hours

• engorge with ingested organisms, necrotic
cells, toxins, Ab-Ag complexes
• often die and lyse (burst)

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 Number
5,000 - 10,000 WBC’s/mm3 blood
– RBC/WBC ratio 700/1
Differential WBC count (a standard clinical lab
report)
– Neutrophils 60-70% of total WBCs
– Lymphocytes 20-25% of total WBCs
– Monocytes 3-8%of total WBCs
– Eosinophils 2-4%of total WBCs
– Basophils 0.5-1% of total WBCs
Abnormal proportions are correlated with different
types of disease processes
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 Leukocyte Number Abnormalities
 Leukocytosis = increased numbers
– Normal component of inflammatory response to injuries and
infections
 Leukopenia = decreased numbers
– malnutrition, chronic disease states
– drug induced - glucocorticoids, anti-cancer drugs, etc.

 Leukemia, Lymphomas = grossly increased numbers, abnormal forms;

many subcategories
– bone marrow and blood stream (leukemia) or tissue spaces
(lymphoma) fill with cancerous (nonfunctional) leukocytes
– crowds out other cells types
• anemia
• bleeding
• immunodeficiency
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 Leukocyte Disorders
 Generally a descendent of a
single cell

– different types of cells

• myelocytic leukemia
• lymphocytic leukemia

– under different cancerous

conditions
• acute - if derived from -
blast type cells
• chronic - if derived from
later stages
 Thrombocytes - Platelets

 Development
– Megakaryocytes shed small cytoplasmic fragments
– Each fragment surrounded by plasma membrane
 Anatomy
– 250,000-400,000/mm3 of blood
– No nucleus, disc shaped
– 2-4 µm diameter with many granules
Thrombocytes - Platelets (cont.)
Physiology
• Short life span (5-9
days)

• Help plug small holes in

blood vessels

• Granules contain
regulatory factors
which serve several
important functions in:
• coagulation
• inflammation
• immune defenses
 Haemoglobin
Hemoglobin molecule consists of two parts:
Haem which contains iron and globin (protein
part).
Haemoglobin synthesis: The main function of
red cells is to carry O2 to the tissues and to
return carbon dioxide (CO2) from the tissues
to the lungs. In order to achieve this gaseous
exchange they contain the specialized protein
haemoglobin.
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 Each molecule of normal adult haemoglobin A
(Hb A) (the dominant haemoglobin in blood after
the age of 3–6 months) consists of four
polypeptide chains, α2β2, each with its own haem
group. Normal adult blood also contains small
quantities of two other haemoglobins: Hb F and
Hb A2. These also contain α chains, but with γ and
δ chains, respectively, instead of β .

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 O2 combines with Hgb in lungs
 O2 gas not very soluble in H2O
 Hemoglobin transports O2 Hemoglobin
– 2 α globin chains & 2 β globin chains
– 4 heme groups (lipid)
– each heme binds an iron ion (Fe²+) that carries 1 O2
 Haemoglobin synthesis

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Haem synthesis
occurs largely in
the mitochondria
by a series of
biochemical
reactions .
protoporphyrin
combines with
iron in the ferrous
(Fe2+) state to
form haem
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 Haemoglobin function :
The red cells in systemic arterial blood carry O2 from the
lungs to the tissues and return in venous blood with CO2 to
the lungs. As the haemoglobin molecule loads and unloads
O2 the individual globin chains move on each other . The
α1β1 and α2β2 contacts stabilize the molecule. When O2 is
unloaded the β chains are pulled apart, permitting entry of
the metabolite 2,3‐diphosphoglycerate (2,3‐DPG) resulting
in a lower affinity of the molecule for O2. This movement is
responsible for the sigmoid form of the haemoglobin O2
dissociation curve. The P50 (i.e. the partial pressure of O2 at
which haemoglobin is half saturated with O2) of normal
blood is 26.6 mmHg. With increased affinity for O2, the
curve shifts to the left (i.e. the P50 falls) while with
decreased
48
affinity for O2, theDr Aydin
curve SA
shifts to the right (i.e.
If you have any
special inquiries
or suggestions
please don’t
hesitate to
aydin@alqalam.edu.iq
discuss them
with me.