Analysis of Uterine Fibroids Using Masson's Trichrome Staining Report

This report examines the application of Masson's trichrome staining in the identification of uterine fibroids. The study begins with an introduction to the Masson's trichrome staining technique, detailing its origins and purpose in histology, emphasizing its use in differentiating tissue structures by producing various colors. The report then outlines the aims, which include using Masson's trichrome to identify fibroids in uterine tissue and understanding the mechanisms of special stains. The methodology describes the step-by-step staining process, including tissue preparation, staining with Weigert's iron hematoxylin, Biebrich scarlet-acid fuchsine, and aniline blue, and subsequent microscopic examination. The results section presents microscopic observations, comparing normal uterine tissue and fibroid tissue stained with Masson's trichrome. A discussion section interprets these findings, explaining the nature of fibroids and the role of Masson's trichrome in visualizing collagen and other tissue components, which is critical in the diagnosis of uterine fibroids and assessment of treatment efficacy. The report concludes by summarizing the importance of tissue stains in diagnosing tumors and the need for a thorough understanding of staining methods.

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Background of the study

Masson’s trichrome (MT) procedure originated from Claude L. Pierre Masson (1880-1959) and
Histology prefers the mentioned three colour staining method. Various colours such as green or blue
collagen, black cell nuclei and pink or light red cytoplasm are produced by most recipe.
Introduction
Staining is a method which is used in diagnosing and studying the abnormal cells morphology e.g.
cancerous cells stressing on the tissues structural components (Gamble and Bancroft). A distinction
between dissimilar structures in a specimen’s tissue is achieved through staining and its examination
is allowed under a light microscope (Cook, 2006). Numerous specimen are diagnosed microscopically
by the use of Haematoxylin and Eosin, haematoxylin helps in staining the nuclei whereas staining of
extracellular materials and cytoplasm is done by the Eosin (Cook and Bancroft, 1995). This method
mentioned is among the main special staining methods used in laboratories and it expresses collective
tissues comprising of extracellular materials and cytoplasm by using three different sized dyes during
staining (Mohr, 2013, p. 178). Masson's trichrome produces three different colours namely; cytoplasm
and muscles which are stained red with Biebrich scarlet, collagen are stained by aniline light green or
blue depending on which one was used.
Aims
 To use Masson's trichrome in the identification of fibroids in the uterine tissue
section.
 To comprehend the mechanism used by special stains.
Method
Masson’s Trichrome
The tissue area surrounded by the paraffin and fixed formalin was rehydrated and deparaffinised
through 100%, alcohol, 95% alcohol and 70% alcohol. The tissue section then washed in distilled
water before staining it in Weigert's iron haematoxylin for 10 minutes (Malvasi, 2014, p. 430).The
distilled water was used again to wash the tissue section after staining it using Biebrich scarlet-acid
fuchsine solution for 15 minutes or until collagen was not red (Mohr, 2013, p. 67) then transferred
straight to aniline blue solution without rinsing and stained for a period of 5-10 minutes . After that,
the distilled water was used to rinse the tissue section for a short time before differentiated in 1%
acetic acid solution for a minute (Goldblum, 2013, p. 78). The washing took place again before a
quick dehydration through 95% ethyl alcohol, which was also used to wipe-off Biebrich scarlet-acid
fuchsine staining. Then the section was mounted for light microscope examination (Harada, 2014, p.
132).
Results
The light microscope was used to carry out the observation and the results below were found

Fig1: 10 x 10 microscopic magnification showing a normal uterine tissue stained with Masson
trchrome. (Harada, 2014)
Fig 2: 10x 10 microscopic magnification picture showing fibroid tissue stained with Masson’s
trichrome (Carretero, 2017)
Discussion
Fibroids also are known as Leiomyomata according to Schorge and Norwitz (2006) are the kind of
myometrium of the uterus that entails extracellular fibrous like collagen and smooth muscle. They

occur in different areas within the uterus but they do not invade the neighbouring organs and tissues
(Lark, 1996). Regular urination, heavy periods, bloating and black ache are its symptoms (Tulandi,
2003).
In figure 1, the uterus tissue stained using Masson’s trichrome exhibited collagen stained in
blue, the nuclei appeared black and erythrocytes and smooth muscles stained red (Mathew, 2013, p.
79). MT discovered that there was a small amount of collagen in the normal uterus shown in figure 1
and also showed a large number of collagen stained in blue which suggested fibrosis in figure 2
(Mathew, 2013, p. 55). The Masson trichrome allows comparison between the level of fibrosis
previously and after treatment to know whether the medication was successful or hence this method is
important in medicine. Collagens are connective tissue proteins adding up to 27. (Goldblum, 2013, p.
99)
Collagens of various types were described by Harada (2013). Whereas their organization were
shown using hydrophilic dye known as Sirius red with polarised light. In Sirius red, three colours
appear i.e. in the first collagen orange or red are seen while the green colour in the third collagen.
(Carretero, 2017, p. 67)
Limitations of Masson’s trichrome
 Blue staining appears faded in case of over differentiation with acetic acid
 Sections which are over fixed in formalin stain poorly
To avoid these problems faced by Masson’s trichrome, it is recommended to use a tissue
section which is normal in size and also avoids over differentiation with acetic acid.
Conclusion
Tissues stains are used widely for diagnosis of tumours of diverse origins. A Proper
understanding of stains methods helps us to carry out our staining procedure successfully and
enhanced innovation of new methods also. The above-mentioned staining methods enable
pathologists in viewing and identifying tissues, microorganisms, and cells.

References
Carretero, J. r. &. A., 2017. Morphological Mouse Phenotyping. 2 ed. Leeds: Readers Digest.
Goldblum, J. R., 2013. Enzinger and Weiss Soft Tissue Tumors. 1 ed. London: University press.
Harada, T., 2014. Endometriosis, Pathogenesis, and Treatment. 3 ed. Chicago: Random House.
Malvasi, A. T. &. A., 2014. Uterine Myoma, Myomectomy and minimally Invasive Treatment. 2 ed.
s.l.:University press.
Mathew, J. N. G. &. R., 2013. The Exstrophy-Epispadias Complex. 3 ed. San Francisco:
ThomsonReuters.
Mohr, U., 2013. International Classification of Rodent Tumors. 1 ed. Birmingham: Random House.