Sridharan Divya, Pracha Nooruddin, Dougherty Julie A, Akhtar Ali, Alvi Syed Baseeruddin, Khan Mahmood
Department of Emergency Medicine, The Ohio State University, Columbus, OH 43210, USA.
Department of Physiology and Cell Biology, The Ohio State University, Columbus, OH 43210, USA.
Methods Protoc. 2022 Jan 27;5(1):13. doi: 10.3390/mps5010013.
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.
马松三色染色法（MTS）是一种通过对组织成分进行差异染色来分析多种疾病病理中纤维化的有用工具。它用于识别心脏、肺、皮肤和肌肉等不同组织中的胶原纤维。特别是在心脏纤维化中，MTS可将胶原纤维染成蓝色，这有助于区分瘢痕区域和健康区域（红色）。然而，使用单一方案对石蜡包埋切片和冷冻切片（冰冻切片）进行染色存在诸多挑战。因此，本研究的目的是开发一种简单的简短方案，以评估石蜡包埋和冷冻心脏切片中的心脏纤维化。MTS使用三种不同的染料，即魏格特铁苏木精、比布里希猩红酸性品红和苯胺蓝，分别用于检测细胞核、细胞质和胶原。在本研究中，我们开发了一种简单的简短方案，任何实验室都可以采用该方案轻松评估石蜡和冷冻心脏切片中的心脏纤维化。此外，我们还解决了免疫染色过程中常见的挑战及故障排除技术。总体而言，我们成功开发了一种简单的一步方案，用于评估石蜡包埋和冷冻冰冻切片中的心肌纤维化。
