DEMONSTRATION OF HAEMATOXYLIN AND EOSIN (H & E) STAINING OF GENERAL TISSUE
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TABLE OF CONTENTS
Pages
Title Page – – – – – – – – i
Certification – – – – – – – iii
Dedication – – – – – – – iv
Acknowledgments – – – – – – v
Table of Contents – – – – – – vi
Abstract – – – – – – – – viii
CHAPTER ONE
INTRODUCTION
1.1 Background of Study – – – – 1
1.2 Aims and Objective of Study – – – 3
1.3 Significant of the Study – – – – 4
1.4 Statement of Purpose – – – – 4
1.5 Statement of Problem – – – – 5
CHAPTER TWO
LITERATURE REVIEW
2.1 Applications of H & E – – – – 9
2.2 Functions of H & E Staining – – – 10
2.3 The Science behind H & E Staining – – 11
2.4 Haematoxylin and Eosin
Staining Solutions – – – – – 12
2.5 Eosins – – – – – – – 14
2.6 Theory of Staining – – – – – 15
2.7 Physical Factors – – – – – 17
2.8 Chemical Factors – – – – – 19
CHAPTER THREE
MATERIALS AND PROCEDURE FOR TISSUE PROCESSING
3.1 Procedure and Timing for Manual
Tissue Processing – – – – – 20
3.3 Constituent of Eosin – – – – – 21
3.4 Preparation of the Stains – – – – 23
3.5 Preparation of Eosin – – – – – 25
CHAPTER FOUR
4.1 Principle of H & E Staining – – – – 26
4.2 Materials for H & E Staining – – – 26
4.3 Procedure for H & E Staining – – – 26
4.4 Result of H & E Staining – – – – 27
CHAPTER FIVE
CONCLUSION AND RECOMMENDATION
References – – – – – – 29
Bibliography – – – – – – 30
ABSTRACT
The haematoxylin-eosin (H & E) stain has stood the test of time as the standard stain for histological examination of general human tissues. However, this simple dye combination is capable of highlighting the fine structures of cells and tissues. H & E does not produce colour randomly, instead the dyes exploit differences in the chemistry of the tissue to differentially colour various components. Haematoxylin and Eosin stain is the standard staining method used in histology, resulting in nuclei being blue-black, cytoplasm giving pink, muscle fibre giving deep red, RBCs giving orange and fibrin giving deep pink. This stain Haematoxylin and eosin is an all purpose stain that is also quick and easy to use which may explain why it has stood the test time.
CHAPTER ONE
INTRODUCTION
1.1 Background of Study
Haematoxylin and Eosin (H & E) staining technique is one of the principal stains and technique in histology. It is the most widely used stains in medical diagnosis and often, the cold standard, for example when a pathologist looks at a biopsy of a suspected cancer. This histological section is likely to be stained with H and E and termed H and E section, it is a combination of Haematoxylin and Eosin which produces different colors based on tissues stained.
Haematoxylin is a dark blue or violet stain that is basic it binds to basophilic substance such as DNA/ RNA which are acidic and negatively charged.
DNA/RNA in the nucleus and RNA in Ribosome’s of the rough endoplasmic reticulum are both acidic because the phosphate backbone of nucleic acids are negatively charged. The negatively charged backbones form salts with basic dye containing positive charges therefore, dyes like haematoxylin will bind to DNA and RNA to give violet colour staining.
Eosin is a red an pink stain that is acidic with negative charge. It binds to acidophilic substances such as Amino Acid side chains e.g. Lysine arginine.
Most proteins in the cytoplasm of some cells are basic because they are positively charged due to arginine and lysine amino acids residues.
These forms salts with acid dyes containing negative charge like eosin. Therefore eosin binds to these amino acids/ proteins and stain them pink. This includes cytoplasmic filaments’ in muscles, cells, intracellular membranes and extracellular fibres.
However, for routine examination, haematoxylin and eosin (H & E) is the stain of choice. The stain produce colours for different tissue structure which would otherwise be transparent, so that one can get a detailed view of the tissue, this two stains were independently introduced in 1865 and 1875 respectively by Bohmer and Fisher. In 1876, Wissowzky described their use in combination as a tissue staining method for staining different materials with different colours.
Despite its simplicity, this stain has stood the test of time. Even now, over a century later, H & E remains the most frequently used tissue stain worldwide.
1.2 Aims and Objective of Study
- To Project the science behind H & E staining
- To Demonstrate the procedure on how to carryout H & E staining.
- To Enlighten us on the need to use H & E for routine tissue analysis.
- To Remind scholars that H & E is the most preferred nuclear stain in histology.
1.3 Significant of the Study
This study will be of immense benefit to histology laboratory personnel’s, as it will enlighten them on the haematoxylin and eosin staining of general tissue and why it is preferred to other staining technique.
This study will also serve as a reference material to other students who might develop interest in the subject matter.
1.4 Statement of Purpose
The main purpose of this research is to investigate and project the importance of Haematoxylin and eosin staining of all tissue (general tissue), also to give the accurate procedure on how to stain tissues with Haematoxylin and eosin for the pathologist to get a clear view of the tissue’s pathological condition.
1.5 Statement of Problem
The amount of information yielded without histological examination is limited, the presence of inflammation, foreign tissues including Trichinella and other parasites neoplasia both primary and metastatic, myofiber, necrosis, perimysial and endothelial connective tissue proliferation and patterns of distribution of atrophic myofibers, subcellular structural abnormalities such as ragged red fibers are subtler and harder to see than with histochemical stain (haematoxylin and eosin). Some such as nemaline rods are almost impossible to detect because of lack of colour contrast but with H & E stain it can be seen easily.
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