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HEMATOLOGY AND CLINICAL PATHOLOGY

(Principles, Procedures and Applications)

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Rapid review of

HEMATOLOGY AND

CLINICAL PATHOLOGY (Principles, Procedures and Applications)

DR SHREYA SHRIVASTAVA

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www.educreation.in

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Dedicated to

My beloved Parents for always being on my side and for their

support and unconditional love.

My guide “Dr. Shreenivas Kallianpur” for his constant

support, supervision and motivation, without whom this work

might not be that much effortless and enjoyable.

My brother for boosting my morale.

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Content List

Sr. No. Content Page

1. INTRODUCTION 1

2. COLLECTION AND HANDLING OF

BLOOD

8

Differences between capillary and

venous blood.

8

Equipment required for specimen

collection.

8

Handling of needle. 11

Vein selection. 14

Measures to improve prominence of

vein.

15

Factors leading to difficult vein conditions.

15

Puncture sites 16

Routine puncture sites. 16

Alternative puncture sites. 17

Applying the tourniquet. 18

Disinfecting the puncture site. 19

Standard procedure 20

Venous blood 20

Capillary (peripheral) blood. 21

Blood collection in neonates and

infants.

24

Helpful hints for blood collection. 28

Safety aspects during blood collection.

28

Procedure after blood collection. 32

Storage of blood. 33

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viii

Serum. 33

Defibrinating whole blood. 33

Cold agglutinins. 34

Anticoagulants used in the haematology laboratory.

34

Effects of storage on blood cell

morphology.

37

3. BASIC HAEMATOLOGICAL TECHNIQUES

40

Quantitative tests 40

Complete blood count. 41

Haemoglobin determination. 49

Erythrocyte sedimentation rate. 70

Spun microhaematocrit. 73

Red blood cell indices. 78

Qualitative tests 87

WBC differential count. 87

4. BLOOD SMEARS 96

Preparation of blood films. 96

Staining blood films. 101

Romanovsky stain. 103

Leishman stain. 105

May-Grunwald-Giemsa stain. 105

Jenner- Giemsa stain. 106

Rapid staining methods. 107

Separation and concentration of

blood cells.

108

Examination of blood films for

parasites.

111

5. REFERENCE RANGES AND NORMAL

VALUES

122

6. CONCLUSION 124

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Introduction

1 | Routine Haematological Tests

Introduction ______________________________________________________

Blood is a bodily fluid in living organisms that delivers

necessary substances such as nutrients and oxygen to the cells and transports metabolic waste products away from those same

cells. When it reaches the lungs, gas exchange occurs when

carbon dioxide is diffused out of the blood into the pulmonary

alveoli and oxygen is diffused into the blood. This oxygenated blood is pumped to the left hand side of the heart in the

pulmonary vein and enters the left atrium. From here it passes

through the mitral valve and taken all around the body by the

aorta.1

COMPOSITION

In vertebrates, it is composed of blood cells suspended in blood

plasma. Plasma which constitutes 55% of body fluid, is mostly water (92% by volume), and contains dissipated proteins,

glucose, mineral ions, hormones, carbon dioxide (plasma being

the main medium for excretory product transportation), and

blood cells themselves.1

Albumin is the main protein in plasma, and it functions to

regulate the colloidal osmotic pressure of blood. The blood cells

are mainly red blood cells and white blood cells, including

leukocytes and platelets. The most abundant cells in vertebrate blood are red blood cells. These contain haemoglobin, an iron

containing protein, which facilitates oxygen transport by

reversibly binding to the respiratory gas. Vertebrate blood is

bright red when its haemoglobin is oxygenated and dark red

when it is deoxygenated.2

By volume, the red blood cells constitute about 45% of

whole blood, the plasma about 54.3% and the white blood cells

about 0.7%.2

Red blood cells contain the blood‟s haemoglobin and distribute oxygen. Mature red blood cells lack a nucleus and

organelles in mammals. The red blood cells (together with

endothelial vessels cells and other cells) are also marked by

glycoprotein that defines the different blood types. The

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Introduction

2 | Routine Haematological Tests

proportion of blood occupied by red blood cells is referred to as

“hematocrit”, and is normally about 45%. The combined

surface area of all red blood cells of the human body would be

roughly 2,000 times as great as the body‟s exterior surface.1

White blood cells are part of the body‟s immune system;

they destroy and remove old or aberrant cells and cellular

debris, as well as attack infectious agents (pathogens) and

foreign substances. The cancer of leukocytes is called

“leukaemia”.1

Thrombocytes also called platelets; they take part in

blood clotting (coagulation). Fibrin from the coagulation

cascade creates a mesh over the platelet plug.1

FUNCTIONS

Blood performs many important functions within the body

including:- 1. Supply of oxygen to tissues (bound to haemoglobin, which

is carried in red cells).

2. Supply of nutrients such as glucose, amino acids, and fatty

acids (dissolved in the blood or bound to plasma proteins-blood lipids).

3. Removal of waste such as carbon dioxide, urea, and lactic

acid.

4. Immunological functions, including circulation of white blood

cells, and detection of foreign material by antibodies. 5. Coagulation, the response to a broken blood vessel, the

conversion of blood from a liquid to a semi-solid gel to stop

bleeding. 6. Messenger functions, including the transport of hormones

and the signalling of tissue damage.2

7. Regulation of body Ph.

8. Regulation of core body temperature.

9. Hydraulic functions.

BLOOD TEST IS DIVIDED INTO TWO PARTS:

HAEMATOLOGY - the measurement of elements of the

blood. BIOCHEMISTRY - the measurement of chemical

substances carried by the blood.

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