Hyaluronan (HA) is a ubiquitous component of the extracellular matrix. The spatial-temporal localization of HA can be visualized in situ using biotinylated HA binding proteins (HABPs). This assay is sensitive to fixation conditions, and there are currently no best practices for HA detection. Thus, the goal of this study was to optimize fixation conditions for visualizing HA in the ovary, kidney, and liver through analysis of six commonly used fixatives for HA detection: Bouin's Solution, Carnoy's Solution, Ethanol-Formalin-Glacial Acetic Acid (EFG), Histochoice, Modified Davidson's Solution, and 10% Neutral Buffered Formalin. Organs were harvested from CB6F1 mice and fixed with one of the identified fixatives. Fixed organs were sectioned, and the HABP assay was performed on sections in parallel. Hematoxylin and eosin staining was also performed to visualize tissue architecture. HABP signal localization and intensity varied between fixatives. EFG and Carnoy's Solution best preserved the HA signal intensity in the ovary and liver, showing HA localization in various sub-organ structures. In the kidney, only Modified Davidson's Solution was less than optimal. Our findings demonstrate that fixation can alter the ability to detect HA in tissue macro- and microstructures, as well as localization in a tissue-specific manner, in situ.

中文翻译：

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为了实现卵巢、肾脏和肝脏中透明质酸的最佳原位可视化，需要组织特异性固定方法。


透明质酸 (HA) 是细胞外基质中普遍存在的成分。 HA 的时空定位可以使用生物素化 HA 结合蛋白 (HABP) 进行原位可视化。该测定对固定条件敏感，目前没有 HA 检测的最佳实践。因此，本研究的目标是通过分析用于 HA 检测的六种常用固定剂：Bouin 溶液、Carnoy 溶液、乙醇-福尔马林-冰醋酸 (EFG)，优化固定条件，以实现卵巢、肾脏和肝脏中 HA 的可视化、Histochoice、改良戴维森溶液和 10% 中性缓冲福尔马林。从 CB6F1 小鼠身上收获器官并用一种已确定的固定剂固定。将固定的器官切片，并在切片上平行进行 HABP 测定。还进行苏木精和伊红染色以可视化组织结构。 HABP 信号定位和强度因固定剂而异。 EFG 和 Carnoy's Solution 最好地保留了卵巢和肝脏中的 HA 信号强度，显示了 HA 在各种亚器官结构中的定位。在肾脏中，只有改良戴维森溶液达不到最佳效果。我们的研究结果表明，固定可以改变检测组织宏观和微观结构中 HA 的能力，以及以组织特异性方式原位定位的能力。