Mucins are of great interest in intestinal research and histochemical methods are often employed to identify them. Since it is in the nature of mucins that they are "hard to hold onto" once they come into contact with water, a frequently used medium in histochemistry, there are a number of challenges that may decrease diagnostic accuracy. As the outcome of methods published for microscopic detection of mucosubstances proved to be unsatisfactory in our hands, the aim was the establishment of a reliable and reproducible protocol. Tissue samples were available from pig feeding experiments. In the present study, we focus on a fixation / staining procedure without making comparisons between differently fed pigs. Several fixation and staining procedures were evaluated for their use in semiautomatic quantification and quality assessment of different mucus fractions simultaneous on one tissue section. Cryostat sectioning, subsequent fixation steps with heat, ethanol and modified Bouin's solution, followed by triple staining with high iron diamine, alcian blue and periodic acid-Schiff turned out to be the best method to identify sulfomucin, sialomucin and neutral mucin simultaneous on one tissue section. This methodology resulted in very good morphology of goblet cells with intact mucin containing vesicles within the cells, which was comparable to ultrastructural electron microscopical observations. Semiautomatic quantification of different mucins was possible. In conclusion, reliable mucus quantification and assessment of mucus quality requires strictly tested procedures. According to our experience, the most important aim after cryosectioning is fast fixation of the mucosubstances, which requires a combination of different fixation steps.

中文翻译：

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猪胃肠道粘液物质：固定，染色和定量。

粘蛋白在肠道研究中非常受关注，通常使用组织化学方法对其进行鉴定。由于粘蛋白的本质是一旦与水（一种在组织化学中经常使用的介质）接触，它们就“很难抓住”，因此存在许多挑战，可能会降低诊断的准确性。由于在我们手中证明了用于显微检测粘膜物质的方法的结果不令人满意，因此目的是建立可靠且可重现的方案。组织样品可从猪饲养实验中获得。在本研究中，我们专注于固定/染色程序，而没有在不同喂养的猪之间进行比较。评估了几种固定和染色程序，以便同时在一个组织切片上对不同粘液成分进行半自动定量和质量评估。低温恒温器切片，随后的加热，乙醇和改良的布因溶液固定步骤，然后用高铁二胺，阿尔辛蓝和高碘酸三重染色，席夫（Schiff）被证明是同时在一个组织上鉴定磺脲类，唾液白蛋白和中性粘蛋白的最佳方法部分。这种方法导致杯状细胞的形态非常好，其中杯状细胞内含有完整的粘蛋白囊泡，这与超微结构电子显微镜观察结果相当。可以对不同粘蛋白进行半自动定量。结论，可靠的粘液定量和粘液质量评估需要经过严格测试的程序。根据我们的经验，冷冻切片后最重要的目标是粘膜物质的快速固定，这需要结合不同的固定步骤。