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...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-01-01
|
| Series: | Methods and Protocols |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2409-9279/5/1/13 |
| _version_ | 1797477624470044672 |
|---|---|
| author | Divya Sridharan Nooruddin Pracha Julie A. Dougherty Ali Akhtar Syed Baseeruddin Alvi Mahmood Khan |
| author_facet | Divya Sridharan Nooruddin Pracha Julie A. Dougherty Ali Akhtar Syed Baseeruddin Alvi Mahmood Khan |
| author_sort | Divya Sridharan |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T21:20:21Z |
| format | Article |
| id | doaj.art-f7e9c638d5dc4051b0f11ca44b7f006f |
| institution | Directory Open Access Journal |
| issn | 2409-9279 |
| language | English |
| last_indexed | 2024-03-09T21:20:21Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Methods and Protocols |
| spelling | doaj.art-f7e9c638d5dc4051b0f11ca44b7f006f2023-11-23T21:24:28ZengMDPI AGMethods and Protocols2409-92792022-01-01511310.3390/mps5010013A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin SectionsDivya Sridharan0Nooruddin Pracha1Julie A. Dougherty2Ali Akhtar3Syed Baseeruddin Alvi4Mahmood Khan5Department of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USADepartment of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USADepartment of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USADepartment of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USADepartment of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USADepartment of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USAMasson’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.https://www.mdpi.com/2409-9279/5/1/13histologyTrichrome Stainingcryogenic tissue sectionsformalin-fixed paraffin-embedded tissues |
| spellingShingle | Divya Sridharan Nooruddin Pracha Julie A. Dougherty Ali Akhtar Syed Baseeruddin Alvi Mahmood Khan A One-Stop Protocol to Assess Myocardial Fibrosis in Frozen and Paraffin Sections Methods and Protocols histology Trichrome Staining cryogenic tissue sections formalin-fixed paraffin-embedded tissues |
| 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 | histology Trichrome Staining cryogenic tissue sections formalin-fixed paraffin-embedded tissues |
| url | https://www.mdpi.com/2409-9279/5/1/13 |
| work_keys_str_mv | AT divyasridharan aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT nooruddinpracha aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT julieadougherty aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT aliakhtar aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT syedbaseeruddinalvi aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT mahmoodkhan aonestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT divyasridharan onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT nooruddinpracha onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT julieadougherty onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT aliakhtar onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT syedbaseeruddinalvi onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections AT mahmoodkhan onestopprotocoltoassessmyocardialfibrosisinfrozenandparaffinsections |