The Significance of Dyes in Identifying Tissue Features

This essay delves into the significance of dyes in identifying normal and impaired histological structures of tissues. It discusses the mechanisms by which dyes attach to tissues, including electrostatic interactions, hydrogen bonding, and Van der Waals forces. The essay highlights the key differences between normal and abnormal tissue features, such as cell size, shape, and growth rate, and explains how these differences are visualized using dyes under a microscope. It emphasizes the importance of understanding tissue structure for disease diagnosis and the role of various staining techniques, including H&E staining. The essay concludes that the application of dyes is crucial for differentiating between normal and impaired tissues, thereby contributing to our understanding of diseases and their underlying cellular and molecular processes. The essay emphasizes the crucial role of dyes in histological and histopathological analysis, and their role in improving the contrast of tissue samples under light microscopy, which is essential for the diagnosis and investigation of various diseases.

Running Head: STAINING-DYES
0
Dyes
Essay
student
6/28/2019

STAINING DYES
1
Dyes in the identification of tissue features
Staining commonly works by applying a dye that stains certain of the constituents of the cell a
light colour, composed with the counterstain that helps in staining the other parts of the body cell
a dissimilar colour. There are a different type of dye can be used to recognize the normal and
impaired, characteristics of the dyes. Two different types of dyes include acidic dyes, and basic
dyes (Dey, 2018). Acidic dyes commonly react with the cationic component s in the cells, on the
other hand, the basic type of dyes react with the anionic constituents of the cells. In the tissue
study, the histological staining is significant in order to observe the structure of the cell,
intracellular and extracellular materials with the help of a microscope. Staining is described as
the auxiliary method applied in the microscopy in order to improve contrast in pictures attained
and to highpoint the arrangements. Stains might be applied to describe and inspect high number
of tissues, cell number or organelles inside specific cells; histological characteristics are valuable
for biological investigation and/or analysis in medication (Bancroft, and Gamble, 2008). In this
particular essay, the significance of dyes in classifying normal and impaired histological
structures of the tissues will be discussed.
The electrostatic mechanism is the common mechanism by which the dyes attaches to the
tissues. The affinity between the opposite ionic groups of the dyes and tissues assist in staining
salt linkage and the ionic attachment are the alternative words to the electrostatic binding. Acids
have negative charges; they bind to the positive charges, particularly the hydrogen (Hawe, Sutter,
and Jiskoot, 2008). In the electrical field, they transfer to the anode side. Tissues with a positive
charge then attract the dyes carrying a negative charge and called acidophilic. The base has a
positive charge, they linked to the negative charges specifically the hydroxyl groups. They
translocate towards the cathode and termed cations. Tissues with negative charge then attract the

STAINING DYES
2
dyes carrying a positive charge and called basophilic. Hydrogen bonding is another mechanism
in which occurs between the dye component and the water (Ribatti, 2018). Water then also
compete for hydrogen attachment sites on the specific tissue. Vander Waals forces are the polar
attractions that are weak and are actually effective for a short distance. The attraction is among
the dipoles, which are the molecules contain both positive and negative charges. For example
staining of the elastic fibres with the help of orcein, where elastic fibre id the hydrophobic
protein and the orcein is a big molecule in addition with the strong dipole. Dye aggregation is the
mechanism in which the molecules of dye have an attraction for each other. The aggregated dyes
can penetrate the less effortlessly in the tissues compared to the dispersed dye (Tseung, Rhett,
Takayama, Wong, and Yuen, Lab Vision Corp, 2011).
Some of the key difference between normal and abnormal features of tissues or cells includes
stickiness, ability to metastasize, shape, maturation, appearance and rate of growth. Normal cells
discharge substances that allow them to stick together; on the other hand, the abnormal cells fail
to secrete these substances. Normal cells remain in their actual place, but abnormal cells lack the
adhesive molecule, this ability to travel in the circulation. Abnormal cells are either very small or
very large compared to the normal cells that tend to possess the same size and shape (Wan, et al.,
2017). The abnormal cells divide and grow rapidly before they are completely mature, on the
other hand, normal cells mature at a specific time. The normal and abnormal cells under a
microscope might look different. The nucleus of the abnormal cells appears both bigger and
darker compare to the normal cells, this is because the cancer cells contain excess DNA
molecule. The normal cell divides themselves and stops later when there are enough cells
produced. But abnormal cells reproduce quickly and keep dividing. The abnormal cells in the
infected tissues acquire the autonomous force to proliferate, they contain inactivated tumour

STAINING DYES
3
suppressor genes, for example, Rb that works to inhibit the cell growth. Impaired cell suppresses
and inactivates the genes and some pathways that normally allow cell death (Reza, and
Iftekharuddin, 2013).
The clinical and biological investigation is reinforced by an understanding of the usual
organization and purpose of the cells and body tissues in addition to the different organs and
arrangements they build up the information the illness in the setting of structure and function
associations allows the difference between the typical tissues and unusual tissues in a specific
illness condition. The difference of these allowed by the recognition and knowledge of the
deviation of usual and impaired histology is extremely helpful in identification of the illness and
their therapeutics (Adeyemo, Akinloye, and Adekanmi, 2017).
Such important restraints of the study are dependent on a piece of detailed knowledge and
capability to recognize the simple types of tissue which associates to make the dissimilar organs
within the body. Having the knowledge of the common tissue structure is important to the
recognition of the changed structure. With the proper understanding of the common histology,
investigators can perceive the types, place and scope of abnormal cells functioned in illness,
whether their inherent morphology is affected representing altered function of the cell, and
whether greater order tissue arrangement is diminished which specifies abnormal functioning of
the organ. Histopathology, alternatively, includes the means to authenticate precise models of
specific illnesses centred on understanding the pictorial representation of the molecular processes
distinguished from usual.
Cells and other components of the tissue are organized in steady identifiable patterns in the
standard healthy condition. Body Tissues commonly have specific describing features, for

STAINING DYES
4
example surface organization and shapes and foundations of the integral cells that are applied in
their recognition and valuation of function. Alterations in these arrangements can be persuaded
by an extensive variety of chemical and the physical encouragements, for example, microbial
contamination and malignancy of the cell in cancer are redirected by the structural modifications
and can be seen under the microscope. Many illnesses for instance Cancer are similarly
characterized by distinctive structural and the chemical irregularities which differ in the usual
pattern of the body issues. This is the foundation of the microscopic investigation of samples.
Inspection of numerous specimens and difference of the structures is confronted as the tissue
segments or smears attained from biopsy processes or aspirations seem dull and not very
comprehensive when observed under the light microscopy (Bancroft, and Cook 1994). This is
why the fixed constituents have a similar grey colour in the preparation that a parallel refractive
index and which sorts it problematic to classify the structure of the tissue. It is important to stain
the body cells/tissues to allow the better visualization of the diverse organizations in conflicting
colours (Gurr, 2012). Staining is mostly carried out by the application of histological-dyes that
are actually the painted organic composites attained from the natural sources such as plants or
from synthetic creation that selectively attaches to or distillate in numerous structures of the cell
and tissue. Dyes comprise auxochromes which are described as the chemical constituents that
allow attachment to the tissue, for example, the–OH group that are ionisable, and chromophores
substances that are used to absorb the observable light accountable for the detected colour
(Sadashiv, et al., 2015). Colour appears when a linked chromophore particle absorbs certain
types of wavelengths of noticeable light. Most contemporary dyes, for instance, Haematoxylin
and the Eosin type of stains frequently used for staining, these stains are produced from the

STAINING DYES
5
simpler organic particles, commonly benzene or its byproducts (Pilling, Henderson, Shanks,
Brown, Clarke, and Gardner, 2017).
Stains are normally targeted as distinctive probes, which have adjustable specificity
contingent on the exceptional ionization or an organic reaction with the structures of tissue and
constituents. Staining considerably does not leads to a casual colouring of the specimens of
tissue, however relatively exploits the alterations in the chemical organization of the tissue
(Tseung et al., 2011). This can be observed by colour dissimilarity contingent on which type of
dye is attached. Colours developed to reflect the actual nature of the body tissues and their
possessions and extend a benefit in the disclosure of particular portions or parts. This allows
thorough visualization of arrangements counting cell structures, for example, the cytoplasm, the
nucleus and different organelles, in addition to extra-cellular constituents. Moreover, under
certain circumstances, for example, glycogen storage illnesses, staining can disclose molecular
complexes and alterations linked with pathological situations (Kumar, and Thirumalesh, 2013).
Improved capability for imagining and recognition of arrangements is the main advantage of
applying the dyes in tissue specimen’s staining. Tissue Staining is consequently plays a serious
part in tissue-dependent diagnosis and investigation permitting the picturing of the morphology
of tissue and characteristics associated with histology, and in differentiating normal and irregular
histological structures. These explanations are adequate to allow examination of tissue condition
and analysis of illness (Alturkistani, Tashkandi, and Mohammedsaleh, 2016).
The most commonly applied staining system is known as H &N which contain two different
dyes; one is haematoxylin and another one is eosin. Staining named Eosin-staining is the acidic
type of dye which is charged negatively and stains basic type of structures in red and pink colour
(Pilling et al., 2017). This specific dye sometimes called eosinophilic. Haematoxylin can be

STAINING DYES
6
described as the basic dye which is obtained from the logwood tree and actually known as
hematein. It is used with the aluminium ions and applied to stain the structures that acid in nature
in purplish-blue colour. Some of the Structures that are stained in purple are the DNA in the cell
nucleus and RNA in the ribosomes. Structures that are stained pink are mostly proteins
presenting within their cytoplasmic filaments available in the cells associated with muscle, the
extracellular fibres, and the membrane present intracellularly (Ruifrok, and Johnston, 2001).
Histological dyes usually applied for staining under the light microscopy comprise the stains that
are H&E (Haematoxylin and Eosin), Van Giessen, and the Masson’s Trichrome, amongst others.
These types of stains are the most usually applied stains for the light microscopy (Venkataraman,
2012). This type of staining is regularly applied as it delivers a very thorough understanding of
the tissue attained by staining the cell arrangements, staining the nuclei with dark blue or purple
color, and the cytoplasm inside cell and the connective tissue in colours of pink. Performing
Staining by applying these and another type of dyes forms a serious part of the analytical picture
specified the adequate contrast attained for the presentation of the morphology of the tissue
(Fischer, Jacobson, Rose, and Zeller, 2008).
In concluding part it is identified that the staining is the important process in the histology and
histopathology as it has different advantages. There are a different type of dyes can be used to
identify the common and unusual features of the tissues such as acidic dyes, histological dyes,
and basic. Some of the mechanisms that help dyes to attach with the tissues include electrostatic
mechanism, hydrogen bonding, and Vander Waals forces. One of its primary benefits of using
dye is being the improvement of contrast amongst dissimilar constituents of the tissue sample,
principally as perceived under the light-microscopy. The complete aim of the histology is to
obtain understanding of regular tissues and different organs, which is important to have the

STAINING DYES
7
information of the changed structure and task of unhealthy cells, tissues and different organs.
Identification of abnormal features of the tissues is also essential for disease identification and
some of the characteristics of the impaired cells include larger or very smaller shape, duplication
before maturation, bigger nucleus, and presence of excess DNA molecules. After reviewing
different articles it can be concluded that the application of dyes to permit for separation between
regular and impaired tissues is important to our knowledge of this.

STAINING DYES
8
References
Adeyemo, S.M., Akinloye, A.J. and Adekanmi, G.B., 2017. The Use of Plant Dyes for Microbial
Staining and Identification: An Eco-friendly and Non-Toxic Alternative Method. Journal of
Advances in Biology & Biotechnology, pp.1-10.
Alturkistani, H.A., Tashkandi, F.M. and Mohammedsaleh, Z.M., 2016. Histological stains: a
literature review and case study. Global journal of health science, 8(3), p.72.
Bancroft, D. and Cook C., (1994). Manual of Histological Techniques and their Diagnostic
Application. PA, USA:
Bancroft, J., and Gamble, M., (2008). Theory and practice of histological techniques. PA, USA:
Elsevier
Dey, P. (2018). Staining Principle and General Procedure of Staining of the Tissue. In Basic and
Advanced Laboratory Techniques in Histopathology and Cytology (pp. 57-67). Springer,
Singapore.
Fischer, A.H., Jacobson, K.A., Rose, J. and Zeller, R., 2008. Hematoxylin and eosin staining of
tissue and cell sections. Cold Spring Harbor Protocols, 2008(5), pp.pdb-prot4986.
Gurr, E., 2012. Synthetic dyes in biology, medicine and chemistry. Elsevier.
Hawe, A., Sutter, M. and Jiskoot, W., 2008. Extrinsic fluorescent dyes as tools for protein
characterization. Pharmaceutical research, 25(7), pp.1487-1499.
Kumar, A. and Thirumalesh, M.B., 2013. Use of dyes in ophthalmology. Journal of Clinical
Ophthalmology and Research, 1(1), p.55.

STAINING DYES
9
Pilling, M.J., Henderson, A., Shanks, J.H., Brown, M.D., Clarke, N.W. and Gardner, P., 2017.
Infrared spectral histopathology using haematoxylin and eosin (H&E) stained glass slides: a
major step forward towards clinical translation. Analyst, 142(8), pp.1258-1268.
Reza, S. and Iftekharuddin, K.M., 2013. Multi-class abnormal brain tissue segmentation using
texture. Multimodal Brain Tumor Segmentation, 38.
Ribatti, D., 2018. The staining of mast cells: a historical overview. International archives of
allergy and immunology, 176(1), pp.55-60.
Ruifrok, A.C. and Johnston, D.A., 2001. Quantification of histochemical staining by color
deconvolution. Analytical and quantitative cytology and histology, 23(4), pp.291-299.
Sadashiv, R., Deshpande, S.K., Chougule, M. and Dixit, U.R., 2015. Haematoxylin and eosin
staining technique using liquid dish washing soap as a dewaxing agent replacing xylene-A
biocompatible technique to demonstrate histoarchitecture of tissue in teaching
laboratories. Medica Innovatica, 4, pp.50-9.
Tseung, K.K., Rhett, N.K., Takayama, G.K., Wong, W.B. and Yuen, D.P., Lab Vision Corp,
2011. Automated tissue staining system and reagent container. U.S. Patent 7,901,941.
Venkataraman, K. ed., 2012. The chemistry of synthetic dyes(Vol. 4). Elsevier.
Wan, T., Cao, J., Chen, J. and Qin, Z., 2017. Automated grading of breast cancer histopathology
using cascaded ensemble with combination of multi-level image features. Neurocomputing, 229,
pp.34-44.