Quantitative analysis of chondroitin sulphate retention by tannic acid during preparation of specimens for electron microscopy.

The Histochemical Journal Pub Date : 1995-06-01
D Levanon, H Stein
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Abstract

The ability of tannic acid to enhance binding of glycosaminoglycans to purified collagen was analysed in an in vitro system using amino sugar analysis on an amino acid analyser, transmission electron microscopy, and scanning electron microscopy. Collagen was purified by digestion with trypsin, papain, and hyaluronidase. Purified collagen was incubated with hyaluronic acid or with chondroitin sulphate glycosaminoglycan and then treated with tannic acid. Tannic acid was found to enhance retention during preparation for electron microscopy of either of the glycosaminoglycans onto collagen fibres. The ability of tannic acid to enhance binding of collagen and glycosaminoglycans might explain, at least in part, its structural reinforcement effect on resected synovial joint-apposing surfaces during preparation for scanning electron microscopy.

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电镜标本制备过程中鞣酸对硫酸软骨素保留量的定量分析。
在体外系统中,使用氨基酸分析仪、透射电子显微镜和扫描电子显微镜对氨基糖进行分析,分析单宁酸增强糖胺聚糖与纯化胶原蛋白结合的能力。胶原蛋白用胰蛋白酶、木瓜蛋白酶和透明质酸酶消化纯化。纯化后的胶原蛋白用透明质酸或硫酸软骨素糖胺聚糖孵育,然后用单宁酸处理。在制备到胶原纤维上的糖胺聚糖时,发现单宁酸增强了保留。单宁酸增强胶原蛋白和糖胺聚糖结合的能力,至少在一定程度上可以解释,在扫描电镜准备过程中,它对切除的滑膜关节表面的结构增强作用。
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