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A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections

Masson’s Trichrome Staining (MTS) is a useful tool for analyzing fibrosis in a plethora of disease pathologies by differential staining of tissue components. It is used to identify collagen fibers in different tissues like heart, lung, skin, and muscles. Especially in cardiac fibrosis, MTS stains th...

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Autores principales: Sridharan, Divya, Pracha, Nooruddin, Dougherty, Julie A., Akhtar, Ali, Alvi, Syed Baseeruddin, Khan, Mahmood
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8877130/
https://www.ncbi.nlm.nih.gov/pubmed/35200529
http://dx.doi.org/10.3390/mps5010013
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author Sridharan, Divya
Pracha, Nooruddin
Dougherty, Julie A.
Akhtar, Ali
Alvi, Syed Baseeruddin
Khan, Mahmood
author_facet Sridharan, Divya
Pracha, Nooruddin
Dougherty, Julie A.
Akhtar, Ali
Alvi, Syed Baseeruddin
Khan, Mahmood
author_sort Sridharan, Divya
collection PubMed
description Masson’s Trichrome Staining (MTS) is a useful tool for analyzing fibrosis in a plethora of disease pathologies by differential staining of tissue components. It is used to identify collagen fibers in different tissues like heart, lung, skin, and muscles. Especially in cardiac fibrosis, MTS stains the collagen fibers (blue color), which helps in the distinction of scar area versus the healthy area (red color). However, there are several challenges to stain both paraffin-embedded sections and frozen (cryosections) using a single protocol. Therefore, the goal of this study was to develop a simple short protocol to assess cardiac fibrosis in both paraffin-embedded and cryo heart sections. MTS uses three different stains, i.e., Weigert’s Iron Hematoxylin, Biebrich scarlet-acid fuchsin, and aniline blue to detect nuclei, cytoplasm, and collagen, respectively. In this study, we developed a simple short protocol that can be adapted by any lab to easily assess cardiac fibrosis in paraffin and frozen heart sections. Furthermore, we have addressed the challenges that are commonly faced during the immunostaining process and troubleshooting techniques. Overall, we have successfully developed a simple one-step protocol to assess myocardial fibrosis in paraffin-embedded and frozen cryosections.
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spelling pubmed-88771302022-02-26 A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections Sridharan, Divya Pracha, Nooruddin Dougherty, Julie A. Akhtar, Ali Alvi, Syed Baseeruddin Khan, Mahmood Methods Protoc Protocol Masson’s Trichrome Staining (MTS) is a useful tool for analyzing fibrosis in a plethora of disease pathologies by differential staining of tissue components. It is used to identify collagen fibers in different tissues like heart, lung, skin, and muscles. Especially in cardiac fibrosis, MTS stains the collagen fibers (blue color), which helps in the distinction of scar area versus the healthy area (red color). However, there are several challenges to stain both paraffin-embedded sections and frozen (cryosections) using a single protocol. Therefore, the goal of this study was to develop a simple short protocol to assess cardiac fibrosis in both paraffin-embedded and cryo heart sections. MTS uses three different stains, i.e., Weigert’s Iron Hematoxylin, Biebrich scarlet-acid fuchsin, and aniline blue to detect nuclei, cytoplasm, and collagen, respectively. In this study, we developed a simple short protocol that can be adapted by any lab to easily assess cardiac fibrosis in paraffin and frozen heart sections. Furthermore, we have addressed the challenges that are commonly faced during the immunostaining process and troubleshooting techniques. Overall, we have successfully developed a simple one-step protocol to assess myocardial fibrosis in paraffin-embedded and frozen cryosections. MDPI 2022-01-27 /pmc/articles/PMC8877130/ /pubmed/35200529 http://dx.doi.org/10.3390/mps5010013 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Sridharan, Divya
Pracha, Nooruddin
Dougherty, Julie A.
Akhtar, Ali
Alvi, Syed Baseeruddin
Khan, Mahmood
A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title_full A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title_fullStr A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title_full_unstemmed A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title_short A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections
title_sort one-stop protocol to assess myocardial fibrosis in frozen and paraffin sections
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8877130/
https://www.ncbi.nlm.nih.gov/pubmed/35200529
http://dx.doi.org/10.3390/mps5010013
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