Basic Essentials

[[p.1]]
Basic Essentials
2.1 ANATOMY AND PHYSIOLOGY OF BLOOD CIRCULATION
Blood is a fluid which is continuously on movement. This process of movement of blood is known
as circulation and the system that sustain the process is known as circulatory system and Dr. William
Harvey was first person to describe circulation in 1616. It consists of mainly the heart and blood vessels
viz. arteries and vein-c-cardiovascular system. The whole system works under supreme control of
nervous system and the cardio-respiratory centre is believed to be located in medulla oblongata or brain
stem. The circulatory activity is also indirectly influenced by some hormones produced in our body.
So it is a cooperative function of nervous and humoral mechanism that coordinate various activities
of heart and blood vessel ensuring proper supply of blood in every comer of our body thereby feeding
the vital organs and tissues with oxygen, nutrient and other essential substances.
Blood vessels are of three types -
arteries, capillaries and veins. Arteries are vessel that carries
blood away from the heart and are having thicker walls consisting Tunica adventitia, media and intima
from outside inward respectively composed of connective tissue, muscular tissue and endothelium.
Artery gradual becomes smaller arterioles and then capillary as they moves away from heart, capillaries
and very small having diameter approximately 7 to 7.5/m and having length 0.2 mm to 0.3 nun capillary
wall consists single layer of endothelium through which interchange between blood and tissue fluid
takes place. As blood flows through capillary wall into the tissue the arterial blood changes into venous
blood after exchange of substance between blood and tissues. This changed blood drains into vein
which is brought back to heart and thence to the lung for purification. So veins are also part of
circulatory system but unlike arteries all the three coats of venous walls contains less elastic and less
muscle tissue and so the veins are less resilient and easily collapses. It also consists of valves to allow
unidirectional flow of blood (Fig. 2.1) towards heart, valves prevents back flow of blood. To ensure
proper supply of blood to any organ two systems of vessels works, one is main or larger system and
the other is subsidiary system of smaller vessels called collaterals and these system again communicates
with each other called anastomosis. Similar system ofcommunication exists in venous side ofcirculation.
Heart (Fig. 2.2) is a hollow muscular organ situated in chest between two lungs and occupies more
on left side of midline. It is placed in a cavity within the thorax known as mediastinum and occupies
anterior part of mediastinal cavity. Heart is a pear-shaped structure weighs approximately 250 to 300
gm and it has anterior, posterior and inferior surface. It has apex and base. Base is broadest part of
heart directed upward and backward, the inferior surface lies over diaphragm anterior surface lies
4

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Basic Essentials
5
From lungs
Left atrium
B
Muscle contracted
••I!
~ ~ To lungs
Pulmonary
veins
Rg. 2.1. Unidirectional blood flow in
vein.
Fig. 2.2. Cross section through heart.
From
lungs
To
lungs
behind sternum and cartilages of ribs. Posteriorly the heart is
in relation with oesophagus. Heart is enclosed in a sac called
pericardium which has two layers namely visceral and parital
layer and a space between them called pericardial cavity.
From outside inward the wall of heart consists of musculature
called epicardium;
myocardium
and
endocardium.
Myocardium is strongest of all
an~
endocardium is a thin
connective tissue membrane lined with endothelium.
Myocardium consists of striated muscle tissue and contracts
invol~tarily,
the valves of ·heart are lined by endothelium.
Heart in human beings consists of four chambers. They are
two atrium 'and two ventricles. Atriums are two, namely right
atrium and left atrium, similarly there is right ventricle and
left ventricle. Both the atrium and both the ventricles are
Muscles relaxed
separated from each other by means of septum normally do
not communicate with each other. It is only atrioventricular
orifice through which atrium and ventricle communicates.
Circulation of blood in heart occurs as follows. Superior
and inferior venacava bringing blood respectively from upper extremity and lower extremity and
coronary sinus bringing blood from walls of heart opens into right atrium. This impure blood now
enters right ventricle from where it passes to lungs through pulmonary artery for purification. The
purified blood from both lungs enters
into left atrium through pulmonary
vein and from where it passes into
left ventricle and thrown into systemic
circulation and coronary circulation
by means of aorta and coronary artery
respectively.
Orifices in the heart are guarded
by folds of endocardium known as
valves which ensures properdirection
of blood flow. Heart has four valves
mainly aortic, mitral, pulmonary and
tricuspid valves and are respectively
placed at the opening of aorta and left
ventricle, left ventricle and left atrium,
pulmonary artery and right ventricle,
right atrium and right ventricle. Mitral
valve has two cusps whereas atrio-
ventricular orifice has three cusps
respectively called bicuspid and
tricuspid valves. Aortic has got three
semilunar valves. These valves
normally prevents back flow of blood.
Heart occupies a position between
third to fifth ribs. Apex is located at

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6
Blood Banking and Transfusion
fifth left intercostal place about 7 to 9 ems away from midline and left border of heart runs obliquely
from apex to upper part of left third cartilage. The right border I to 2 cm away from midline run from
third right coastal cartilage to the fifth right coastal cartilage on right side of sternum.
Heart produces its action through rhythmic contraction and relaxation respectively known as
systoles and diastole of atria and ventricles. During systole contraction of heart occurs which diminishes
in size and thereby squeezes out blood from respective chamber i.e. from atria to ventricle and then
simultaneous contraction of both ventricles from where blood is forced into aorta and pulmonary trunk
and these actions of atrias and ventricles are known as phase I and phase II of cardiac activity, in phase
III atria and ventricle relaxes. All these three phases constitutes a single cardiac cycles. Phase I lasts
for 0.1 second, Phase II lasts for 0.3 seconds and Phase III lasts for 0.4 seconds. So it requires 0.8
second for a cardiac cycle to be completed. The contraction of the heart per minute is known as heart
rate and varies with age, physical status etc., normally it ranges between 60 to 80 per minute at rest;
in new born or infant, 120 to 140 per minute and in aged, 90 to 100 per minute. Stress condition and
physical exercise increases the heart rate. The amount of blood discharged by ventricle in one minute
is referred to as cardiac output and is the product of stroke volume (60 mI, i.e, blood ejected per
contraction) and number of cardiac contraction per minute. Heart sounds are produced due to action
of valves i.e, closure of valves. It can be detected by means of stethescope which distinguishes the
sounds in first and second sound. First sound is due to ventricular systole i.e. ventricular contraction
and closure of atrioventricular valves known as systolic sound. The second sound is due to closure
of semilunar valves during the ventricular diastole called diastolic sound. Normally the above two
sounds are heard but in case of pathological conditions other adventitious sounds are also heard viz.
murmur etc.
Circulation of blood in human body consists mainly of two circuit, systemic or greater circulation
and pulmonary or lesser circulation. Systemic circulation starts with aorta that arises from left ventricle.
Aorta divides into arteries which are further subdivided in arterioles, and as they enter different organ
further subdivide into capillaries. From capillaries blood pass into venules which joins to form smaller
veins and they in tum joins to form larger veins. From all veins of upper ·extremity the blood drains
into superior vena cava and those from lower extremity drains into inferior vena cava and both open
into the right atrium of hea.t,
Aorta is the largest of all arteries in our body and it has three parts namely arch of aorta, ascending
aorta and descending aorta. The ascending aorta as it leaves the left ventricle and within the pericardium
gives off right and left coronary artery that supplies the blood to heart. From the arch of aorta which
is continuation of ascending aorta, arises inorninate artery, left common carotid and left subclavian
arranged from right to left. Innominate further. subdivides into right common carotid and right subclavian
and supplies right half of head, and right upper extremity; similarly left common carotid, left subclavian
supplies left side of head, neck and left upper extremity.
Common carotids on each side divides into internal carotid and external carotid. The internal
carotid enters into cranial cavity to supply blood to brain and its structure, to eyeball, lacrimal gland
and skin over forehead. The external carotid is the second branch supplying the head, neck, face,
tongue, pharynx, jaws, teeth and muscle of mastication on the corresponding sides through its different
branches. Subclavian artery supplies mammary gland, thyroid, larynx two upper intercoastal space and
muscles of occiput. Vertebral artery considered to be largest of all the branches of subclavian artery
and both joins to form basilar artery and along with branches from internal carotid forms circle of
Willis at base of brain. Axillary artery continuation of subclavian artery'supplies upper limb through
brachial, radial and ulnar artery respectively in upper arm, forearm and hand of corresponding sides
i.e. left and right upper limb. Descending aorta continues down as thoracic aorta and abdominal aorta.

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Basic Essentials
7
The thoracic aorta supplies the organs in thoracic cavity, walls of thorax and diaphragm through its
visceral and parital branches also known as intcrcostal artery. After that thoracic aorta passes through
a orifice in diaphragm and continues as abdominal aorta which has visceral and parital part, supplies
the abdomen and its content. Abdominal aorta at the level of fourth lumbar vertebra divides into right
and left common illiac which further subdivides into internal illiac and external illiac supplying the
true pelvis and its content and then supplies the lower limb.
Pulmonary or lesser circulation begins with pulmonary trunk that arises from right ventricle and
carries impure blood to lungs and after purification the blood is brought back to left atrium through
pulmonary veins. Pulmonary artery in lungs divides into arterioles and capillaries which spreads over
aveoli where exchange of gas between inhaled air and venous blood takes place and in this process
blood receives oxygen and gets rid of carbondioxide, In tissue where oxygen is consumed by cells
and intercellular substance and in tum liberates carbon dioxide which is carried by venous 'blood into
lungs. The gaseous interchange in lung is governed by definite physical law that the pressure of a gas
in liquid differs from that of gas in surrounding air, the gas passes from liquid to air and vice versa
till the pressure are equalised. Blood is the media via which carbon dioxide and oxygen is transported.
Oxygen dissolves in plasma of blood and combines with haemoglobin of R.B.C. and forms an unstable
element known as oxyhaemoglobin, about 90 to 96% of inspired oxygen in blood is converted into
oxyhaemoglobin and gaseous interchange takes place in tissue where oxygen is released and carbon
dioxide passes into blood from surrounding tissue fluid and carried over to the lungs in venous blood
as carboxyhaemoglobin to be exchanged with oxygen in the inhaled air. This purified blood is brought
back through venules and pulmonary vein into the left atrium.
Capillaries are minute blood vessels visible under microscope and consists of single layer of
endothelium, the interchange of gases and other substances between blood and tissue fluid takes place.
This interchange of nutrients, gases and other substances through capillary wall is dependent not only
on the capillary wall but also on the connective tissue surrounding the walls of the capillary. In state
of rest most of the capillary remains contracted as such blood flow is diminished or nil. The blood
as flows through capillary, its composition changes and it becomes venous blood and then drain into
venules which in tum drain into vein.
Venous system consists of portal, pulmonary and systemic circulation. The portal system is situated
in abdominal cavity. It collects venous blood from stomach" small intestine, large intestine, spleen,
pancreas and gall bladder. The portal vein enters the liver through porta hepatis, subdivides in smaller
branches similar to a capillary network. Blood than pass into central hepatic vein and along with other
hepatic veins forms a common vein to open into inferior vena cava; the lesser or pulmonary venous
system has been described earlier. Systemic circulation consists of inferior vena cava system and
superior vena cava system. In inferior vena cava system the vena cava passes through orifice in
diaphragm into the thoracic cavity drains into right atrium and inferior vena cava is formed by junction
of two common illiac veins, the common illiac veins are formed by junction of internal and external
illiac vein respectively, they drains blood from corresponding half of true pelvis and entire leg.
Superficial vein begins on the dorsum of the foot runs up along medial side to open into femoral vein
in thigh another system of superficial vein located on posterior side runs up to join the popliteal vein.
Thus inferior vena cava system drains blood from abdomen and its content and both lower lirnbs..in
fact it drains the whole lower half below diaphragm.
Superior vena cava is located in the anterior mediastinum. It drains blood from upper extremity
including head, neck and thoracic cavity through azygos, internal jugular, and subclavian vein. The
internal jugular drains blood from the corresponding half of head and neck region and subclavian vein
drains or receives blood from arm, shoulder and part of the neck. Internal jugular and subclavian joins

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8
Blood Banking and Transfusion
Fig. 2.3. Cubital fossa content and position.
~~'
.~---
Pronator teres
-nuscie
.'1.t~~
-
-Basilic
vein
.......~i'-i-
Bicipital aponeurosis
~...........
Accessory ulnar vein
__.,,:;;.;;j..- Ulnar artery
~ ...........:?-
Deep communicating
"ein
Medial cutaneous
-," .c.>'...........-
Nerve of forearm
Basilic vein
Median nerve
'l;.niiri'Wl~
Brachialis muscle
;,.;;~:-.-
Brachial artery
=f-....:;..;.;-.....-
Median basilic vein
Cephalic vein
Radial artery
- _ ~_
..;:
Biceps Muscle -
--;;"""-';;.--- ,
Cephalic vein -
-
-+I
Brachialis muscle -
-
..=-.
Median vein of forearm -
......,
-
i- ~
""1
Radial
Recurrent artery
Lateral cutaneous _
.........;;.
Nerve of forearm
Median cephalic vein • -
,;......,
P:'-.
..;,;r
'ti!
~ !:
:.
Accessory cephalic vein
Brachio-radlalis muscle --,.o..-j--
to form Brachiocephalic vein on each side and both brachiocephalic joins to form superior vena cava
which finally open into right atrium.
The upper limb has two systems of veins (a) deep veins, (b) superficial veins. The deep veins
are paired and placed by the side of similarly named arteries. There is frequent communication between
deep and superficial veins. The deep veins are tightly bound by fascia and surrounding muscle which
prevents them from collapsing unlike the superficial veins which collapses easily. Superficial veins
are mainly cephalic and basilic veins which are connected by median cubital vein at the region of
antecubital fossa where all these veins are placed very much superficially.
Basilic vein begins at medial end of dorsal venous arch and receives dorsal digital branch from little
finger ascends medially and upward, near the elbow it passes infront the medial epicondyle. Here it is
joined by median cubital vein, it further ascends up in the upper arm where it drains into brachial vein.
Cephalic vein begins at radial end of dorsal venous arch it receives two dorsal digital veins from
thumb
and then turns around the radial border of distal end of foreannand ascends upward to the
front of elbow where it is connected by median cubital to basilic vein. It then further ascends up to
drain in to axillary vein.
The brachial and axillary vein drains into subclavian vein of corresponding side and finally opens
into superior vena cava.
Median cubital vein is the large communicating vein between cephalic and basilic vein situated
superficially infront of elbow in the region of cubital fossa (Fig. 2.3) and is separated from brachial
artery deeply placed under the bicipital aponeurosis. So bicipital aponeurosis separates the brachial
artery and median cubital vein which is placed superficially in the cubital fossa. Cubital fossa is a
hollow infront of the elbow triangular in outline, base being an imaginary line drawn between two
epicondyle, medial border by pronetor teres muscle and lateral border by brachioradialis muscle and
both meets at the apex. Roof
is formed by expansion of
tendinous insertion of biceps,
strengthened by deep fascia
which is known as bicipital
aponeurosis, this is pierced by
communication between deep
vein and median cubital vein.
The fossa contains a quantity
of fat, termination of brachial
artery and proximal part of
radial and ulnar artery, tendon
of biceps brachi, lateral side
of brachial artery and median
nerve on medial side of artery,
anterior cutaneous nerve of
forearm and lateral cutaneous
nerve of forearm.
Arterial system of upper
limb consist of axillary artery
which is continuation of
subclavian artery. Axillary

[[p.6]]
Basic Essentials
9
artery continue downward as brachial artery in the upper arm passing downward and laterally and
enters the cubital fossa where it divides into radial and ulnar artery. The ulnar artery larger of two
courses down and medially becomes superficial at the region of wrist, in palm it form superficial palmar
arch and gives off a deep palmar branch. Radial artery smaller of two passes downward and laterally
and near the wrist it becomes superficial and is placed lateral to flexor carpii radialis. Here the radial
pulse is counted. In palm radial artery takes part in formation of deep arch. From both the palmar
arches arises the digital arteries which supplies the fingers and' are placed on either side of fingers
and opposite the terminal phalanx both branches joins together to form arch which supplies pulp of
finger and nail bed etc.
The arterial and venous system of ann, forearm and hand is important for blood bankers and
transfusionist because:
1. Superficial veins in anticubital region are the commonly used ones for collection and
administration of blood, also samples are collected from these veins
2. Reading of blood pressure is taken in upper arm
3. Radial pulse is felt over wrist
4. Skin puncture site at fore finger, middle finger and ring finger for collection of blood samples
for specimen.
Lymphatic system
Consists of lymph vessels and lymph node and circulating lymph. It can be considered as an additional
outflow system supplementing the venous system. Composition of lymph is similar to blood plasma
but in addition it contain lymphocyte in suspension. The lymph from all the lymph vessels drains into
two largest lymph vessels thoracic duct and right lymphatic duct respectively opens into left venous
angle formed by junction of left subclavian and internal jugular and right venous angle fanned by
junction of right subclavian and internal jugular. Drainage area for right lymphatic .duct is very small
viz. right half of thorax right upper extremity, right half of head, face and neck. Drainage area for
thoracic duct is the rest area of whole body excluding those drained by right lymphatic duct thoracic
duct also drains small intestine and as such the lymph in the thoracic duct contains fat droplets and
may be other nutrients.
Foetal circulation
The foetus receives nutrients and oxygen from mother through placenta and through placenta only
eliminates the waste product. Inflow of blood is through umbilical vein i.e, from placenta to foetus
and outflow is through umbilical artery i.e. from foetus to placenta, umbilical vein divided in two one
extends to liver and ductus venosus opens directly into inferior vena cava and so inferior vena caval
blood is a mixture of arterial and venous blood. Inferior vena cava enters right atrium thus the blood
passes through foramen ovale into left atrium and then to left ventricle and aorta. Superior vena caval
blood enters right atrium, the right ventricle and pulmonary trunk and since lung is non-functioning,
greater part of blood passes through ductus anteriosus into arch of aorta and it also contain mixed
blood. So blood passes through descending aorta has less oxygen and that ascending aorta and branches
from arch contains more oxygen. Umbilical artery is the branch arising frora internal illiac artery, it
carries blood from foetus to placenta. After birth cord gets disconnected from placenta due to tieing
and cutting, lungs opens up spontaneously the breathing starts. Forman ovale, intra atrial septum,
ductus arteriosus and ductus venosus gets obliterated into ligaments in due course.

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10
Blood Banking and Transfusion
2.2 BLOOD
Word blood is related to Blowan means bloom or flourish. Blood is a vital fluid of our body and as
such the life line of human body. It is a red coloured viscid fluid slightly salty in taste. Blood is alkaline
in reaction pH 7.4 and specific gravity ranges from 1.052 to 1.060. In adult human, blood volume
ranges between 4.5 to 6.0 liters and is approximately about one thirteenth of adult human body weight.
Temperature of circulating blood is 37.7°C. Blood has two main components, cells and plasma. Cells
consists of 40 to 45% of the total amount of blood and plasma consists of 55 to 60% of total amount
of blood, cells are the formed elements and are of three types i.e. red cells (erythrocyte), white cells
(leucocyte), platelets (thrombocyte) and each has its own characteristic described in 'Table 2.1.
Table 2.1. Cellular elements
Red CeUs
Total count-4.5 to 6.2 million/ul, They are biconcave discs, 7.2 ~ (7 to 8 rang) average
diameter and 1.7 ~ (1.5 to 1.7 range) is average thickness. Life span 120 days. They are flexible
and distensible can pass through narrow capillary. Average surface area of red cell is 120 mu
and volume is 85 mm and this large ratio of surface area and volume is responsible for gaseous
diffusion. Total surface area of circulating red cells is 1500 times of body surface area and
a red cell travels approximately 170 miles in its life span of 120 days to deliver oxygen to
tissues.
RBC contains haemoglobin responsible for maintainance of tissue respiration by transporting
oxygen in form of oxyhaemoglobin to tissues and carbon dioxide from tissue to lung in form
of carboxy haemoglobin. Haem stands for iron and globin stands for protein parrofhaemoglobin
molecules (Fig. 2.4C).
Lymphocyte
Basophil
Monocyte
B. Platelet
Neurophil
Eosinophil
A. Mature leucocyte
side view of a red cell
C. Redcel!
Ag. 2.4. A. Mature leucocyte; B. Platelet; C. Red cell.

[[p.8]]
Basic Essentials
11
Fig. 2.5. Blood smear different
cellular element.
N utrophilic
Band cell
Basophilic
Segmentedcell
Lymphocyte
Neutrophilic
Segmented cell
Plat lets
Eosinophili
Segmenad cell
Total count--4.000 to 11.000 per ul of blood,
Unlike red cells the leucocytes are nucleated.
They are actively
mo{~1e
varies in shape. In
general the chemical composition of leucocyte
82% of its content is water. It carries
nucleoprotein. intracellular enzymes and
measurable traces of inorganic contents sodium.
potassium, calcium chloride. phosphate and
bicarbonate. Lymphocyte contains y globulin.
Monocyte and granulocyte contains enzyme. In
the cytoplasm ofleucocytes granules arepresent
and on this basis they are divided in granular.
Average size: 9 to 12 J.I. further subdivided on
basis of staining reaction :
(a) Neutrophils-Fine dust like granules in
cytoplasm stains light violetmyelocyte 0.3%,
juvenile 9%, Neutrophil-Band 0 to 5%
segmented neutrophil 40 to 60% (Fig. 2.5).
(b) Eosinophil 1 to 3% (contains large granules
of equal size stains pink).
'
(c) Basophil 0 to 4% (contains granules of
different size stainblue).
Non-granular (no granules in cytoplasm) :
(a) Lymphocyte-Size 6.5 to 8.5 J.I. in diameter large round nucleus, thin and blue cytoplasm.
(b) Monocyte count is 2040%, 12 to 20 J.I. in diameter with bean-shaped nucleus, 4 to 8%
(Fig. 2.4A).
White Cell
Platelets
Total count-2oo to 400 thousand per microlitre of blood. size 2 to 3 J.I. and irregular in shape.
Life span 10 days. Secretes enzyme thrombokinase main function clot fonnation platelets are
negatively charged contains acid phosphomonoesterase, ascorbic acid, catalase dopaoxidase,
proteolytic enzyme lipids. Cholesterolesterase, natural fat, RNA, thrombo plastin etc. (Fig.
2.4B) and (Fig. 2.5).
Main function of leucocytes are protective through phagocytosis.
Origin and development of blood cells consists of three successive stages in foetal life and they
are mesoblastic haemopoiesis, hepatic haemopoiesis and myeloid haemopoiesis.
It was Bloom in 1940 who first observed blood cells of human embryo in yolk sac. Gelmour in
1941 and Knoll in 1949 described subsequent maturation. Mesoblastic haemopoises demonstrable
during first two months of embryonic life consists of primitive. erythroblast ~
normoblast (small).
Hepatic haemopoieses during second to fifth month when spleen and thymus also takes part in blood
formation. Small erythroblast appears which has very less cytoplasm and coarse chromatin.'Myeloid
haemopoiesis starts at fifth month, corresponds to establishment of placental circulation. At this stage
liver is concerned with erythropoisis and marrow leuopoises. Later on marrow takes over. Though the
fixed mesenchymal cells are reduced to scant stroma, it retain the haematopoitic potentiality throughout
life.
In post natal life erythrocytes, granulocyte, monocytes and platelets are produced and lymphocytes
in thymus, secondary lymphoid organs and bone marrow.The process is a complex one where

[[p.9]]
12
Blood Banking and Transfusion
·pleuripotentia) stem cell jn the process of evolution differentiate into committed progenitor cells, they
in turn proliferate into functional and matured cells as described below (Table 2.2). The committed
progenitor cells are glycoprotein in nature and they are granulocyte macrophage colony stimulating
factor (GMCSF), granulocyte colony stimulating factor (CGSF) then interlukin 3 (IL-3) and many
others.
Table 2.2. Bone marrow
CPu
Basophil
!IL3
Basophil
I
Eo-CFC
I
GCSF
GMCSF
Myeloblast
I
Eosinophil
Metamyelocyte
I
Eosinophil
I
CFu-N
I
Pro
Myelocyte
I
Neutrophil
I
GMCFC
I
Myeloblast
I
I
CFu-M
r
Pro
Monocyte
I
Monocyte
IL3
GMCSF
GCSF
I
BFu-MEG
IIL3
CFu-MEG
I
Pro
Megak-
aryocyte
Meta
Megakaryocyte
I
Platelet
I
BFu-E
GMCSF
ILl
IL2
CFu-E
I
Prononno-
blast
I
Reticulocyte
I
Erythrocyte
ILl
IL2
GMCSF
BONE MARROW
I
--------- Pleuripotential-------- Myeloid
stem cell
stem cell
I
T cell
Thymus
I
Pre T
Interleukin
Burst forming unit
Colony forming unit
Colony forming cell
Eosinophil-CFC
Erythroid
Monocyte
Neutrophil
Granulocyte macrophage colony stimulating factor'
Granulocyte CSF
Lymphoid
stem cell
~
IL
BFu
CFu
CFC
EO-CFC
E
M
N
GMCSF
GCSF
Bone
f
Pre-,::
Pre B
llA
IL5
ll..6
B cell
Plasma
Plasma consists of 55 to 60% of human blood volume and is slightly yellowish protein fluid. It is a
viscous medium in which the formed or cellular element remain suspended. In plasma 90 to 92% is
water and rest 8 to 10% is formed of organic and inorganic substances, organic substances are mainly
protein viz. Albumin, Globulin and Fibrinogen. It also contains fat, sugar in form of glucose, amino

[[p.10]]
Basic Essentials
13
acids, enzymes and hormones. Inorganic substances consists of 0.9 to 1% of blood plasma, they are
found
a~ salts of calcium, potassium and sodium also the metabolic products found in plasma are in
form of urea and uric acid (Table 2.3).
Main function of plasma is :
1. Carrying metabolic and carbondioxide from tissues
2.. Protective through antibody found in plasma
Main function of blood are as follows :
1. Metabolic--carriage of nutrition to tissue and bring back the waste products from tissue for
excreation.
2. Protective function is exerted through antibody and phagocytosis.
3. Hormonal regulation by transporting hormone to different parts of body.
4. Temperature regulation by dissipitating temperature from hotter to cool area.
5. Respiratory function by carrying oxygen to tissues and bring back carbon dioxide from tissue
to lungs.
Prqperty of blood
Physically blood is red coloured viscid fluid containing R.B.C., W.B.C., platelet and plasma, the
oxyhaemoglobin in suspension is responsible for its scarlet red colour whereas less oxygenated venous
blood is dark red. The opacity of blood is due to different refractive power possessed by the RBCs
and plasma. Blood becomes transparent when RBCs are haemolysed and cannot reflect light. The other
physical property of blood includes colloid osmotic pressure, colloid stability, property ofsedimentation,
viscosity, specific gravity, surface tension, absorption of radiation, rotation of polarised light and light
scattering.
Osmotic pressure of blood is being maintained largely by kidney. This value for whole blood
corresponds to an osmotic pressure of 7.9 atmosphere at bodytemperature. The solutions having equal
osmotic pressure as that of blood is called isosmotic viz. 0.9% sodium chloride solution, osmotic effect
of blood is due to dissolved electrolytes, responsible for crystalloid osmotic pressure and plasma
protein responsible for colloid osmotic pressure. Vascular membranes are not permeable to proteins.
Direction of flow of fluid as such will be from low hydrostatic pressure to that of high osmotic pressure
of plasma at venous end and vice
ve~a.
This results in equal osmolality in both sides of membrane
separating the compartment.
Table 2.3. 9omposition of plasma
Diffusable
Non-diffusable
J. Anabolic-glucose, amino acid creatine
1. Protem - Albumin, Globulin.
Fibrinogen
2. Catabolic - urea, uric acid and creatinine
2. Lipid
+
+
++
++
3. Electrolytes - Na , K
t Ca
,Mg
, Cl , RC03, HP04' H2P04
3. Enzyme
4. Hormones and vitamins
4. Antibody

[[p.11]]
14
Blood Banking and Transfusion
Colloid stability
Serum stability depends on hydration and the charges of its protein molecule. Progressive dehydration
with ageing is accompanied by changes in colloids. Serum stability decreases in acute conditions of
liver and kidney and joint.
Sedimentation
Red cells remains uniformly distributed in plasma throughout the blood which is continuously under
movement in vivo. But in vitro when cells are allowed to stand without any movement, the cells comes
together and settles down leaving clear supernatant plasma. Process is known as sedimentation and
is influenced by electrical charges on red cells which is normally negative, so any condition accelerating
positive charges in plasma cause changes in sedimentation rate viz. level of oxygen, cholesterol and
fibrinogen whereas it decreases in elevated level of CO 2 and albumin.
"Viscosity
The resistance of blood to flow is due to friction developing between the cells and protein suspended
in the plasma. The viscosity of blood is thus connected with peripheral resistance and therefore, to
the >blood pressure in artery. Viscosity depicted by 11 is the ratio of shear stress to rate of strain. Viscosity
of blood in man varies from 4.4 to 4.7 nun. It is influencedby whole body hydration, body temperature,
number of cells present which is usually constant.
Temperature of blood
Blood in body is constantly under movement and thereby dissipates heat from hotter to cooler under
regulated control frorn hypothalamus. This system if required delays the blood flow through skin
facilitating heat through evaporation, radiation, conduction and convection. Regulatory mechanism
helps in heat preservation by constricting the skin vasculature and inducing shivering by muscle
contraction.
Specific gravity
The specific gravity of whole blood varies between 1.052 to 1.060. It is between 1.089 to 1.097 for
RBC and 1.024 to 1.027 'for plasma and is determined by falling drop method at room temperature.
A drop of blood is allowed to fall through CuSO4 solution and specific gravity can be determined
through rise and fall of drop of blood which reveals specific gravity in relation to CuS04 solution.
Specific gravity is influenced by cell plasma ratio.
Electrical conductivity and zeta potential
Conductivity of blood varies inversly with the number of cells since it offers resistance to passage
of charged ions in the stream of blood.
Erythrocytes has negative charge present on the cell surface and since like charges repels, the
erythrocyte in solution remains apart from each other approximately at a distance of 25 nm.
The degree of negative charge is expressed at zeta potential. It is the potential difference between
red cell surface charge and cation in the medium. Cations are again of two types those attached to
the cells and those unattached. Boundary of shear is the boundary between these two cations. Zeta
potential is determined at boundary of shear and expressed in -mv. Pollack in 1970 observed that it
is possible to alter the electrosta,tically imposed intercellular distance by increasing the dielectric

[[p.12]]
Fig. 2.6. Primary plate11et
plug.
A
A. Platelet plug inside a opened
blood vessel
Basic Essentials
15
constant of medium thereby reducing the zeta potential viz. colloid-
B.S. albumin and by the efficiency of a particular diluent to change
the physical environment e.g. low ionic strength solution.
The most important property of blood that deserves special
mention is property of coagulation. The classical concept of
coagulation is described by Schmidt in 1893 that explains the
qualitative phenomenon and remains unchanged despite the
quantitative experiment which measures the activity of various new
components playing part in the process ofcoagulation. In coagulation
of blood thromboplastin, prothrombin, thrombin, fibrinogen and fibrin
plays the vital role. The arrest of bleeding depends upon primary
platelet plug formation (Fig. 2.6) in conjunction with elaboration of
a stable fibrin clot and this process involves a series of plasma
protein acting in a sequential order and highly complex manner.
The essential steps are preparatory stage -7 conversion stage -7
polymerisation stage.
In preparatory phase blood thromboplastin generation takes place
which is influenced by tissue factor and water wettable surface.
Disintegration of platelet
liberating
thromboplastic
and
vasoconstrictive compound, activation of plasma thromboplastin
content occurs. Second step is conversion of prothrombin to thrombin. In this step thromboplastin react
with prothrombin and reaction is catalysed by calcium and platelet factor 1.Third step is polymerisation
ofFibrinogen. Thrombin now acts on Fibrinogen converts it into soluble Fibrin monomer and polypeptides
and subsequently into polymerised fibrin clot through catalysis by calcium and Fibrin stabilising factor
(for details larger text book on haematology to be consulted). The pertinent question that automatically
arises is regarding the maintenance of haemofluidity. The components that promotes the cascading
reaction of blood coagulation are counteracted by inhibitors that maintains the fluidity of blood. If
rate of formation of coagulation component is more then rate of inactivation by inhibitors; then the
coagulation will result. Amongst the physiological inhibitors the antithrombin III is the most important
and it mainly inhibits the thrombin and factor Xa. It has been further observed that antithrombin III
can also inhibit factors XIIa, XIa, IXa. The Ct i.e. plasma inhibitor of complement has inhibitory
activity against XIIa, XIa and Kallikrin. Protein C circulates in blood in inactivated form activated
by thrombin has inhibitory action on activated factor VII and V. In normal fibrinolysis the preformed
clot responsible for arrest of bleeding is dissolved through fibrinolytic system that involves a complex
phenomenon and in short described as follows. Plasminogen circulating in inactivated form is converted
into plasmin. The bonds in fibrin molecules are cleaved sequentially by the plasmin resulting in release
of peptidic substance called Fibrin degradation product, FOP thus produced has anticoagulant effect
and inhibits Fibrin polymerisation and also acts against thrombin. The process is very complex and
beyond the scope of this book.
Amongst all the major constituent of blood red cell is more important from blood bankers point
of view because it fulfills approximately the 80% of transfusion needs in form of packed red cells.
The W.B.C. platelet and plasma are considered dangerous potentially and thus have restricted use.
Further red cell through its haemoglobin does the most important of function of oxygen carriage.
Red cell is composed of 63% water, 33% is haemoglobin, 0.5% lipids and 3.5% is mixture of sugar,
salts, enzyme, protein, amino acid and vitaminsetc. Electron microscopyof red cells reveals thatcytoplasm
is homogenous containing autophagic vacuole and degenerated particles.Stromatin is intracellularstroma.

[[p.13]]
16
Blood Banking and Transfusion
-----+)
Coproporphyrin
Porphobilinogen
---~)
uroporphyrine
---~)
Haeme
B. Two molecules of oALA
C. 4 molecules of porphobilinogen
---~)
Protoporphyrin
D. Protoporphyrin + Iron
Erythrocyte has membrane and consists of four layers. The outer most layer is of sialoglycoprotein and
is a discontinuous layer, this glycoprotein appears to have originated from within the membrane and
consists of ultrastructure of radially oriented lipid and tangentially oriented protein, ABO and RH group
antigenic determinant are primarily glycoprotein and placed on outer layer of red cell membrane. Second
and third layer is made of lipids-cholesterol, lecithin, phospholipid, glycolipid etc. Their presence on
outer surface determines the osmotic property of red cells which preserves protein and electrolyte for its
internal metabolism. Non-electrolyte enters red cell via lipid solubility. Dextrose permeates readily and
acts as a source .of energy but lactose, sucrose and maltose cannot penetrate. The energy derived from
glycolysis where passage of ions are required. Sodium and chloride are pumped out into the surrounding
plasma while retaining potein, phosphate and potassium. Fourth layer is innermost layerconsist ofspectrin
tetramer joined by actin filaments and is concerned with ATPase activity. Most important function of red
cell is oxygen carriage which it does as follows : •
Inspired air P02 158 mm ~
alveolar air P02 103 mm ~
arterial blood P02 94 mm ~
lymph
P02 30 nun ~
tissue cell P02 10 mm, oxygen flows through vasculature in alveoli and mixes with
plasma most of which diffuses across the membrane and combines with haemoglobin. Haemoglobin
is a tetramer and its polypeptide chain contains haeme linked with globin. Haeme in reduced state
binds oxygen. Finally it is carried into circulation andto tissues where oxygen tension is low and the
oxygen is released. Oxygen release is thus dependent on oxygen tension. If oxygen saturation at a
given tension is low then oxygen is released but if it is more less oxygen is released to the tissue.
Experimentally it can be plotted in curve known as sigmoid curve. In former situation curve shifts
to left. Oxygen carriage is also influenced by shape of cells which are flexible and deformable that
allows it.s easy passage through capillary. Further.2-3 DPQ ATP, Nicotinamide adenine nucleotide,
Nicotinamide adenine dinucleotide phosphate and glutathione reductase are the five metabolic
intermediary formed during anaerobic conversion of glucose to lactate. The anaerobic glycolysis helps
in oxygen transport bycontroUing oxygen affinity (2-3 DPG) and membrane function (ATP). Haeme
is maintained in a reduced state and denaturation ofhaemoglobin prevented respectively by nicotinamide
adenine nucleotide and nicotinamide adenine dinucleotide phosphate.
Carbon dioxide also is transported by blood, the flow from tissue is influenced by pressure gradient.
Tissue PC02 50 mm ~
venous blood PCOz 46 mm ~
alveolar air PC02 is 40 mm ~
expired
air PC02 32 mID ~
atmospheric air PC02 0.3 nun. The blood carries its large reservoir of CO2,
combined as NaHC03 and carbaminoprotein in plasma and as carboxyhaemoglobin and KCH03 in
RBC. Since much of the cell K is bound by haemoglobin and acid radicals the total .C02 content of
.plasma is three times that of RBC. The integration of above two system shows the reaction involved
in lungs and tissue. The' PC02 in venous blood is higher and P02 is lower then alveolar air, hence
C92 diffuses from blood Into lung for exhalation and oxygen diffuses for oxyhaemoglobin formation.
In tissue P02 - 90 mm is higher in arterial blood and PC02 is 40 mm which is lower than in tissue
and hence CO2 flows from tissues to blood raising PC02 in venous blood.
Haemoglobin is the major constituent of RBC and approximately forms about 33%. Haemoglobin
is formed in erythrocyte precursor i.e, Normoblasts. It has two moiety namely Haeme and globin.
Haeme synthesis occurs as follows :
A. Glycine + Succinyl Co-enzyme A ----t) Delta aminolevulinic acid'
Ala dehydrase

[[p.14]]
Basic Essentials
17
Globin polypeptide chains are formed in cytoplasm of normoblast and reticulocyte. Specific small
soluble RNA and coded messenger DNA takes part in formation of polypeptide chains which when
released from ribosomes folds into a three dimensional structure.
Control of Haemoglobin synthesis is 'very complex and beyond the scope of this book.
Normal adult haemoglobin, therefore, is a tetramer of four haeme group linked to two pairs of
unlike polypeptide chains. Each polypeptide chains has a haeme group linked to the globin at a specific
site Haeme in a reduced state can bind oxygen. As the partial pressure of oxygen increases four haeme
group sequentially binds one molecule of oxygen Haemoglobin taking part in transport of oxygen and
CO2 has been described earlier.
Red cells has certain property which is in difference to other formed element of blood and they
are in relevance to blood banking.
Sedimentation
Erythrocytes having higher specific gravity then plasma settles down in vitro where RBCs are not in
motion, the rate at which RBC settles down is normally 4 to 10 mm per hour and this is known as
erythrocytic sedimentation rate.
Rouleaux formation
This is piling up of red cells one upon another so that they resembles like piled up coin. In wet
preparation it is difficult to distinguish between rouleaux .and agglutination. In fact rouleaux gets
dispqtsed when a drop of saline is added but not the agglutination. Elevated plasma protein.
par¥roteinimia, addition of high molecular dextran may all cause rouleaux formation. The red cells
also exhibits certain other properties under condition of immune response viz..Agglutination, HaemoJysis
etc. discussed in appropriate chapter.
2.3 GENES AND INHERITANCE
In an individual all inherited characteristic are carried by genes (Genos is a Greek word means offspring
or race) which are bits of nucleic acid occupying a discrete region along the length of chromosome.
Chromosomes are thread-like structure found in the nucleus of a cell and are divided in autosomes
and sex chromosomes. In human being there are 22 pairs of autosome and one pair sex chromosome
in female it is XX and in male it is XY. During meiotic cell division homologus paired chromosomes
parts i.e, 23 pairs parts to become 23 chromosomes in each nucleus of ovum and sperm. Thus when
zygote is formed each chromosome finds its homolog during fusion of ovum and sperm and since every
individual carries two genes for each attribute, therefore, the genetic information are normally not lost
or gained as individual receives 23 chromosome from maternal side and 23 chromosome from paternal
side and the contained genetic information is passed away from one generation to other but then it
follows some established principles formulated by Gregor Mendel-mendelian theory of inheritance.
A very short summary is as follows :
1. Hereditary traits are passed from one generation to the next through some particles genes-
law of unit character.
2. A pair of g~nes
for each attribute is present in an individual. Splitting of these genes occur
at the stage of garnet formation and they segregate and only one goes to the ovum the other
goes to sperm-law of segregation.

[[p.15]]
18
Blood Banking and Transfusion
3. Genes occur in pairs for a particular trait and it may be an unlike pair and one may mask
the expression of other. The character that expresses itself is called dominant and the character
that does not appear in generation is called recessive-law of dominance.
4. The assortment of a pair of gene is independent of the other. Presence of any gene in a single
garnet thus results from random distribution of gene present in the concerned species-law of
independent assortment.
5. Results can be expressed in terms of mathematical probability for mating occurring between
the members of opposite sex of the same species-law of mathematical probability.
The genes are stable but in the long process of evolution spaced between generation it may slowly
undergo some changes for a given characteristic spontaneously-mutation, which may give rise to some
alternate forms for each of the genes and are termed as allele or allelomorphic gene. A system in which
two or more alleles are found for a particular attribute, there exists a variety and is referred to as
polymorphic. The position on a chromosome occupied by a gene is called locus and when identical
allele occupy two loci of a paired chromosome the person/individual is called homozygous and when
unidentical allele contributed by each parent occupies the two loci, the person is said to be heterozygous.
Total genetic constitution of an individual is referred to as genotype and outward expression of genetic
constitution of an individual is referred to as phenotype. The phenotype expression may mark the
presence of other allele in some cases. The distinguished allele in such cases are called as dominant
and the suppressed or masked one is known as recessive. This applies for both homozygous and
heterozygous states. The product of recessive gene can only be observed when the person is homozygous
for appropriate recessive gene. In co-dominance each gene expresses its character equally particularly
in heterozygous. The gene which complies with genetic definition of recessive genes or does not
produce any recognisable effect are termed as silent gene or amorphus, e.g., in ABO system of blood
group 0 does not give rise to any enzymatic activity and hence amorph. Homozygous state can be
differentiated from heterozygous state only in blood group serology e.g. DD cells will react more
strongly with antisera-0 then the heterozygous Dd which will usually show poor strength of reaction.
This variance in expression of antigen observed in vitro is termed as "Dosage effect". Again the genes
when placed very close to another in the same chromosome, called linked genes and are usually
inherited together but when situated at a distance apart are known as unlinked and are .inherited as
independent and it has been also observed that loci far apart has a greater chance of cross over i.e.
exchange of segments may occur between the homologous paired chromosomes and by counting these
gene combination in offspring, one can calculate the frequency of cross over. The reassortment of genes
in a garnet to accommodate the parental genes is termed as recombination and cross over frequency
or recombination fraction :.; expressed in centemorgan, So centemorgan is expression of frequency
in per cent, e.g. recombination frequency of 2% reflects that two loci are situated at a genetic distance
of 2 em from one another. Blood groups are inherited characteristic, the blood group precursors and
their biochemistry is also under genetic· control and follows normal genetic dictum and will be
discussed in appropriate chapter.
2.4 SPECIMEN COLLECTION
Blood and its component are most frequently analysed fluid required in connection with blood banking.
The blood can be collected by puncturing the vessels carrying them i.e. vein and artery and also it
can be collected from skin by skin puncture.
Blood is collected mostly from vein for investigative purposes and rarely from artery. Vein is
selected for the purpose of withdrawal of sample (commonly used site is anti-cubital fossa) is anchored

[[p.16]]
Basic Essentials
19
Pisiform
Common flexor sheath
Flexor sheaths -J. ~ ~~F~
=r n l
of fingers
Finger pric~
site .L.
~---j
~-1
hrl~
+-
Tubercle of scaphoid
Sheath of flexor pollicis longus
Fig. 2.7. Blood specimen collection site from finger pricks
are done only on pulp of distal phalanx index, ring and
middle.
and toumiqet applied. Necessary asceptic
precaution is taken. Disposable needle and
syringe is used for the purpose. After
proximal application of toumiqet the needle
is inserted into the vein appropriate quantity
of blood is collected either in plain tube or
EDTA.
Arterial blood is very rarely required
for investigative purpose except for
measurement of pH, CO2 and 02 tension.
In plain tube 5 to 7 ml of blood is
collected for following investigative
purpose. The plain vial or tube after
collection of sample is properly stoppered
and labelled by a brown paper containing
information such as name, age, sex, ward,
bed number and statistical number of the
patient, the informations may be typed or
hand written.
Investigations are :
I. Indirect and direct coombs test
2. Antibody detection and screening
3. ABO grouping and Rh grouping
and cross-matching
4. Antinatal evaluation
5. Post-transfusion reaction
6. Serum is separated from collected
blood in plain tube and used for
investigation of HCV, STS, HIV,
HbSAg - 2 ml serum for each.
Blood collected in plain tube is devoid of any anticoagulant. This blood yields serum on centrifugation
or sedimentation or by use of serum separator tube. In contrast the anticoagulated blood yields plasma
that contains fibrinogen.
Blood specimen are collected in anticoagulants sometimes and the commonly used anticoagulant
is Ethylene diamine tetra acetic acid (EDTA). The EDTA has anticomplementary activity and used
in following investigation viz. :
(a) Investigations in cases of AIHA where it prevents autoimmune haemolysis of blood samples.
(b) Screening of alloantibody.
(c) In some monoclonal reagent to prevent haemolysin activity of fresh serum.
The other anticoagulants are ACD and CPD used for preservation and storage of blood. Heparin
is used for sample collection in investigations connected with HLA system.
Blood can also be collected by skin puncture method from forefinger, ring finger and middle finger
in case of adults and from heels (medial and lateral planter surface) in case of children and infants.
The blood collected by skin puncture is a mixture of venous, arterial and capillary blood in contrast

[[p.17]]
20
Blood Banking and Transfusion
to popular belief that it is a' capillary blood. Disposable needle should be used for finger prick or
heelprick after necessary asceptic precaution is observed, firstdrop of blood isdisca.ded and then the
part again is cleaned with sterile cotton then second drop can be collected on slide or micropipette
can be usedto aspirate blood varying from 1 to 200 ul, oral aspiration is avoided at all cost. Skin
puncture technique is used in cases of bums, obese patients, for outdoor blood grouping, estimation
of haemoglobin etc. Conunon precautions to be taken :
(a) Oral aspiration is to be avoided at all cost.
(b) Milking of site of collection is avoided for obvious reason of haemolysis,
(c) Proper identification of samples in case specimen received along with blood requisition form.
(d) Specimen collected in plain tubes should not be preserved for a long time continuously
because contamination and loss of antigenecity occurs, to avoid this loss of antigenecity,
sample can promptly be stored at (-) 20°C.
(e) According to Drug and Cosmetics Rules samples of blood received alongwith requisition form
are to be preserved for seven days and for donor's blood it is to be preserved for three days.
(f) Mouth of vial to be properly stoppered to prevent contamination or drying.
(g) While receiving sample ensure that the sample is not contaminated or haemolysed.
(h) Release tourniquet as soon as blood starts flowing.
(i) Do not bend the withdrawal site, use only light pressure application by cotton wool after
withdrawal completed.
Collection of urine sample
Examination of urine is also required in connection with investigation of blood transfusion reaction
and they are :
(i)
Investigation of transfusion reaction to find out presence of RBC, billirubin and WBC.
(ii)
Culture and sensitivity.
Random specimen are collected in clean receptacle during post-transfusion reaction period, labelled
properly.
For culture purpose it is collected in a sterile vial meant for the purpose and avoid contamination
especially microbial, morning urine is preferred for this investigation.
Processing
Serum specimen can be processed by sedimentation or centrifugation of blood collected in plain vial
supernatant part is serum and it is devoid of fibrinogen. Centrifugation is usually done at 1000 to 2000
RPM for three minutes.
Plasma specimen can be processed by centrifugation of anti-coagulated blood supernatant is
plasma and contains fibrinogen.
RBC specimen is processed from the bottom layer of sedimented or centrifuged blood collected
in plain vial and cell suspension is made with addition of normal saline viz. 5% cell suspension means
5 drops of RBC in 95 drops of normal saline (0.90/0 NaCI). Pooled RBC is prepared by preferably
pooling three different cell samples of freshly collected blood of appropriate group. This pooled cell
samples are then made into appropriate cell suspension and can be kept in the rack for use. At room
temperature these pooled samples starts getting deteriorated by six hours so if required fresh samples

[[p.18]]
Basic Essentials
21
Table 2.4
Volulne (liquidmeasure)
-1
(a) Decilitre 10
litre
-3
(b) Millilitre 10
of litre
-6
(e) Microlitre 10
of litre
-9
(d) Nanolitre 10
of litre
-12
(e) Picolitre 10
of litre
-15
(f) Femtolitre 10
of litre
3
(g) One litre 10
Weight
-3
(a) Milligram 10
of gram
-6
(b) MicrogramlO
of gram
....9
(e) Nanogram
1~12
of gram
(d) Picogram 10
of gram
3
(e) One kilogram 10 of gram
Unit of substance is expressed in mole (mol)
.
-3
(a) Millimole (mmol) 10
-6
(b) Micromole (umol) 10
Length (linear measure)
-1
(a) Decimeter 10
of meter .
-2
(b) Centimeter 10
of meter
-3
(e) Millimeter 10
of meter
-6
(d) Micrometer 10 of meter
-9
(e) Nanometer 10
ofmeter
3
(f) One meter 10
Concentration is expressed in moleJIitre and thus substance concentration' will be expressed as :
-3
(a) Millimolellitre= 10 nitre
-6
(b) Micromolellitre = 10 nitre
Similarly mass concentration is expressed in gram/litre (gil)
-3
(a) Milligramllitre = 10 nitre
-6
(b) Microgramllitre = 10 nitre
are to be prepared again viz. for B group. It is required to collect appropriate amount of cells from
plain vial sample of freshly collected blood of B' group and three such samples.are mixed thoroughly
to prepare B pooled cells.
2.5 UNITS OF MEASUREMENT
Basically required in blood group serology and related topics are briefly enlisted as below. The units
can be broadly divided in volume, length and weight for the purpose of convenience. The volume is
expressed in litre, the length is expressed in meter and the weight is expressed in grams. The above
three units are commonly used. Area which is expressed in square is not used in blood group serology.
2.6 BASIC EQUIPMENTS
Before going in to the functioning details we must have some knowledge of basic blood bank
equipment required for smooth running of blood bank.

[[p.19]]
22
Blood Banking and Transfusion
Basic blood bank equipment reagents and chemicals
A. General equipment
I. Diesel generating set (capacity 6 kVA)
2. Exhaust fan (heavy duty)
3. Air conditioner (capacity 1 tonne)
4. Voltage stabiliser 16AH
5. Voltage stabiliser 12 AH
6. Universal standard room thermometer
7. Emergency lights(minimum 3 tubes)
B. Listof laboratory equipments
1. Walk in cooler - 5890 litre
2. Temperature recorder and alarming system in walk-in-cooler
3. Refrigerator - 265 litre
4. Binocular research microscope
5. Monocular research microscope
6. Bacterological incubator oven (455 x 455 x 455 mm)
7. Circulating fan -for oven
8. Bacteriological incubator (455 x 455 x 455 mm)
9. Laboratory centrifuge
10. Water bath 56°C
11. VDRL rotator
12. Distilled water still (cap 5 litrelhour)
13. Thermograph recorder (electrical) for autoclave
14. Vertical steam steriliser with 6 kW heater (400 x 600 mm model)
15. Elisa readers for hepatitis Band HIV
16. Migration cell
17. Ultraviolet hood 4 x 1Y2 X 1Y2
18. Vacuum pressure pump
19. Centrifuge machine (manual)
20. Weighing machine
C. Laboratory instrumentsand donor room material
1. Blood administration set
2. Blood donor set
3. Plasma aspiration set
4. Adhesive plaster
5. Cotton absorbant
6. Foreceps plain
7. Foreceps artery
8. Foreceps (cheatle)
9. Haemoglobinometer

[[p.20]]
Ba~c
Essentia~
23
10. Needles (26 G) disposable
11. Needles (21 G) disposable
12.
B.~
instrument table type
13. B.~
instrument wall type
14. Sthethoscope
15. Thermometer wall type
16. Rh viewing box
17. Plastic blood bag (single, double, tripple) with anticoagulants or blood bottles where
bottles arein use.
General: Polythene bottles (250 ml, 500 ml, 1000 ml, 2000 ml), beaker (small, medium and
large - plastic), brush for cleaning tube, enamel tray with lid. Plastic bags-coloured, colourless,
brass rack for tubes, wooden stick for grouping and crossmatching lit, 1", wooden stand for
tubes, rubber teat-big and small forpipettes, tourniquet, wire basket for tubes, gauze lint,
numbering machine for blood bottles in case bottles are used for blood collection, test tubes
(assorted), glass slide.
D. Laboratory glassware: Evaluated container, glass bottle (100, 500 and 1000 ml), glass bottle
wash, Pasteur pipette, graduated pipette - 0.1, 1.0, 2.0, 5.0 ml, syringe - 2.5 to 10 ml, glass
slides - 500 pieces, test tubes - 50 x 50 ml (100), 75 x 10 ml (100),4" x ~ ml, 6" x ~ ml, coplin
jar vertical, measuring cylinder (1000), measuring cylinder (500 ml), glass capillary (100).
E. Reagents and chemicals: Antisera -A, -B, -AB, -D, antihumanglobulin, anti Al (lectin) serum,
ACD (B) solution, normal saline, denatured spirit, Australia antibody +ve serum, Australia
antigen +ve serum, human plasma (lyophilised), bovine albumin 20% (100 ml), elisakits for
1llV, Hepatitis B, Hepatitis C screening.
F. General Ammonia solution, dettol, de-ionised water, litmus paper - wide range, copper sulphate
solution (sp. gravity 1053).
G
Chemicals forAustralia antigen testing - Sodium azide, Sodium acetate, Sodium diethyl barbitrate,
Egarose.
H. Chemical for papain cystin enzyme preparation - cystin Monohydrochloride, Papain powder,
Potassium dihydrogen, Di-sodium hydrogen phosphate D, IN Hydrochloric acid.
I. Emergency drugs - Inj. Atropin, Inj. Adrenalin, Inj. Avil, Inj. dexona, Inj. Gluconate, Inj.
Dextrose Saline bottles, Oxygen cylinder with accessory.
The following are further added to the list :
1. Ice box (portable for carrying/transport of blood from camps and other places).
2. CPD solution now widely used as anticoagulant in bags.
3. Microtitre plate for grouping where such method is used in laboratory.
4. Hand gloves.
N.B. : All needles and syringes used must be of disposable nature.
Description of certain important equipment
1. Serological water bath : Double walled inner chamber made up of polished stainless steel,
with special glass wool insulation in between. Immersion heating element fitted at bottom,
drain plug for emptying water, stirrer provided for continuous stirring for ensuring uniform
distribu-tion of temperature throughout. Temperature control from ambient to 100°C by solid

[[p.21]]
24
Blood Banking and Transfusion
state electronic temperature controller cum
indicator. Table top model is suitable. 37°C
temperature is required for serological work
and 56°C is required for complement
.destruction (Fig. 2.8).
2. Lab. centrifuge: Used for centrifugation
of samples in laboratory fitted with :
Speed indicator: Variable speed up to 5000
rpm.
G Value : 3500
Swing out rotor: Capacity to be deter-mined
as per requirement
Automatic timer: 0 t060 minutes. Size to '----
-
----J
be determined
Fig. 2.8. Serological water bath.
Electric dynamic brake and fuse safety
device
.Different models (Fig. 2.9) are available
may be selected according to individual
requirement.
3. Hot air oven : Used for heating, drying
andsterilising the instruments, glassware etc.
t>ouble walled inner chamber is made up of
stainless steel or Aluminium, outer wall of
S.S. with 65 mm gap inbetween filled with
special grade glass wool. Inner wall to have
ribs for 213 removable shelves. Fitted with
spring loaded door closing device and heavy
hingeswith ball catcher system. Temperature
ambient to 300°C fitted with capillary type
thermostat; with air circulating fan, timer
and digital indicator cum controller
electronically operated (Fig. 2.10).
Blood bank refrigerator : It is different
from other refrigerator in the sense that it
has to constantly maintain temperature
between 4 to 6°C for preservation and '--
_
storage of blood. So any fluctuation in
Fig. 2.9. labOratory centrifuge.
current can not be allowed and current has
to be regular and stabilised, e.g. 230 V, 50 Hz at 30 amp. Fitted with fan for air circulation
automatic audio-visual indicator for current, automatic defrosting. Shelves for four blood
groups fitted with tray divided in two for positive and negative groups in each shelf. Capacity
according to individual requirement (Fig. 2.11)..·
5. Rh view box: It is a box with small tube light fitted inside, controlledby regulator to increase
or decrease the light, the temperature is preferably maintained at 37°C. White milky glass is

[[p.22]]
Basic Essentials
- 25
FIg. 2.10. Hot air oven.
Fig. 2.11. Blood bank refrigerator.
B.
Open showing tray with blood bags
fitted infront of the box covering the tube.
Slide is kept over the glass cover and
viewed after switching on the light.
6. Shaker for VDRL and microtitre plate
: Has aplatform at the top to accommodate
VORL plate or microtitre plate for
shaking, that moves in anticlockwise
motion, automatic timer - 0 to 60 minutes,
speed indicator with adjustable speed from
o to 180 rpm. G .value is 100.
7. Autoclave : High pressure water steam,
electrically heated. Horizontal steriliser
used for sterilising plastic material, rubber
and injection liquids etc. by means of
steam under pressure 10 to 20 psi
(adjustable). Electrical supply 440 volt, 3
pH, 50 Hz supply of AC current.
Double walled chamber and boiler both
mounted on heavy steel, inner wall of
stainless steel. Radial lockingsystem with
automatic control and self closing device.
8. Thawing bath : This is used for rapid
and uniform thawing of frozen plasma
and red cell units, capacity is variable. It
can also be used for thawing of red cells
units and takes approximately about 10-
12 minutes. Safety thermostat are provided
to protect plasma; red cells or any other
product
for
thawing
from
over
temperature. Automatic timer ring
(adjustable - 30 minutes) is also provided
to indicate completion of thawing.
Stainless steel filter screen to protect the
pump from debris. Full water circulation
for uniform thawing and drainage system
are inbuilt.
Accessories such as compression rack
holder, thermometer; there is also flat
freeze rack for holding frozen plasma and
CIO:~
view
red cell cannister rack holder for frozen
L-
--:.
..::...--:-
-.J
red cell unit.
9. Walk-in-cooler : It is a chamber fully
airconditioned to maintain temperature between 4 to 6°C where an operator can .walk in
whenever required. Fitted with central audiovisual monitoring system, shelves of perforated

[[p.23]]
26
Blood Banking and Transfusion
Rg. 2.12. Spring bafance.
13.
stainless steel four in number for four groups of
blood and divided in middle to accommodate
positive and negative group.
10. Deep freezer : Maintains temperature between
-30°C to -7(J'C, mainly for deep freezing of
plasma and products and other blood components.
11. Platelet incubator : Platelets are preserved at
room temperaturei.e. +22°C and it has been
further observed that platelets preserved at +22OC
are more effective than the platelets storedat +4OC.
Also at room temperature of +22°C platelets
remain viable for five days maximum but up to
72 hours the viability is best maintained.
The incubators may be single/dual type, it holds platelet rotator and agitator. It has arrangement
for air circulation with high density insulation to maintain uniform temperature. Temperature
can be fixed at 220C or kept variable as per range the minimum starts from +15OC. Alarm
system included. Chart recorder for temperature recording. Interior to be made up of stainless
steel. Audio-visual alarm system should include high low safety alarm system.
12. Spring balance : This is used for weighing the blood collected in the bag from the donor
so that we know the exact amount that is collected and stop the bleeding of donor after actual
collection. The blood bag is hangedwith the hook of the spring balance. Each ml of blood
approximately weighs 1.05 gm so 350 ml will measure 367 gm an amount of ± 10% is allowed
(Fig. 2.12).
Air incubator : Some busy laboratory where large scale serological work is undertaken
prefers air incubator to water bath to provide a temperature of 37OC. A hot room at 370C has
added advantage to accommodate bench, microscope and
centrifuge etc.
Recommended for further reading
1. Biochemistry ofBlood in Health and Disease I, Newton Kuglemass, first edition, 1959, publisher, Charles
C. Thomas, Bannerstone House, 301-327, East lawrence Avenue, Springfield Illinois; USA.
2.
Cardiovascular Physiology -
By Robert M. Berme and Mathew N. Levey, third edition, 1977, C.V. Mosby
and Company.
3.
ClOInighams Manual of Practical Anatomy (Vol. I), 12th edition, Oxford University Press, London.
4. Gray's Human Anatomy, 34th edition; 1967 by Dr. D.V. Davies, Univ: of London.
5. Introduction to Transfusion Medicine, Zarin S. Bharucha and D.M. Chouhan.
6. Human Anatomy and Physiology, V. Tatarinov; Fourth edition, 1978, Mir Publisher; Moscow.
7. National Guide Book on Bl~od
Donor Motivation, First edition, 1990, Published by DGHS, MOH and
F.w.. New oau-uoon.
8.
Samson .Wright's Applied Physiology, 12th edition, 1971, Revised by Cyril A. Keele and Eric Neill, ELBS
and Oxford University Press.
9. An Introduction to Medical Genetics by J.A.F. Robert's and Marcous E. Pembrey; Seventh edition, 1978,
ELBS and Oxford University Press.
10. Genetics in Medicine by LS, Thompson and M.W. Thompson, Reprinted edition, Nov. 1970, W.B. Sounder's
and Company, Philadelphia and London.