Picrosirius Red Staining: Revisiting Its Application to the Qualitative and Quantitative Assessment of Collagen Type I and Type III in Tendon
Picrosirius Red Staining: Revisiting Its Application to the Qualitative and Quantitative Assessment of Collagen Type I and Type III in Tendon
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DOI：
10.1369/00221554211046777
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发表时间：
2021-09-22
影响因子：
3.2
通讯作者：
Mueller, Sebastian A.
中科院分区：
文献类型：
--
作者：
De Padilla, Consuelo M. Lopez;Coenen, Michael J.;Mueller, Sebastian A.
胶原蛋白在肌腱的结构组织中起主要作用。在偏振光显微镜下观察的天狼星红（PSR）染色是评价组织中胶原纤维组织化的标准方法。它还用于区分组织切片中的I型和III型胶原蛋白。然而，由于技术因素和历史误解，PSR图像的准确分析和解释具有挑战性。本研究的目的是澄清是否可以区分胶原I型和III型PSR染色在大鼠跟腱，使用双重免疫组化作为阳性对照。我们的研究结果表明，PSR染色观察偏振光显微镜是适合定性和定量评估的总胶原，但不能区分胶原类型。我们发现使用配备旋转台的偏光显微镜至关重要;相对于交叉偏振器呈45度角的肌腱切片方向对于胶原组织的定性和定量评估是最佳的。免疫组化检测III型胶原上级优于PSR染色。我们还比较了福尔马林和布因溶液作为固定剂。两者产生相似的双折射，但福尔马林固定的肌腱提供了更高质量的苏木精-伊红和免疫染色的组织学细节：
Collagen has a major role in the structural organization of tendons. Picrosirius red (PSR) staining viewed under polarized light microscopy is the standard method to evaluate the organization of collagen fibers in tissues. It is also used to distinguish between type I and type III collagen in tissue sections. However, accurate analysis and interpretation of PSR images are challenging because of technical factors and historical misconceptions. The aim of this study was to clarify whether collagen types I and III can be distinguished by PSR staining in rat Achilles tendons, using double immunohistochemistry as the positive control. Our findings showed that PSR staining viewed with polarized light microscopy was suitable for qualitative and quantitative assessment of total collagen but was not able to distinguish collagen types. We found it critical to use a polarizing microscope equipped with a rotating stage; tendon section orientation at 45 degrees with respect to crossed polarizers was optimal for the qualitative and quantitative assessment of collagen organization. Immunohistochemistry was superior to PSR staining for detection of collagen type III. We also compared formalin and Bouin solution as fixatives. Both produced similar birefringence, but formalin-fixed tendons provided higher quality histological detail with both hematoxylin-eosin and immunostaining: