THIONIN STAINING OF PARAFFIN AND PLASTIC EMBEDDED SECTIONS OF CARTILAGE
THIONIN STAINING OF PARAFFIN AND PLASTIC EMBEDDED SECTIONS OF CARTILAGE
复制标题
DOI：
10.3109/10520299309105572
复制
发表时间：
1993-01-01
影响因子：
1.6
通讯作者：
VANDERLINDEN, AJ
中科院分区：
文献类型：
--
作者：
BULSTRA, SK;DRUKKER, J;VANDERLINDEN, AJ
硫堇的有用性染色的软骨切片包埋在乙二醇甲基丙烯酸酯（GMA）和脱钙的影响，包埋在石蜡和GMA的软骨切片进行了评估。使用5%甲酸或10%EDTA的短脱钙时间不影响染色性质或软骨基质和软骨细胞的形态。用于软骨差异染色的标准染色剂番红O-坚牢绿色在这些实验中用作对照。番红O染色切片长时间暴露于固绿色导致番红O染色基质消失，从而妨碍带负电荷的糖胺聚糖（GAG）的定量测量。硫堇染色均匀，在整个软骨层，独立的染色时间。与番红O相反，硫堇在非脱水软骨切片中没有表现出异染性，这使得它更适合于评估GMA嵌入软骨中的软骨质量。为了评价软骨中硫堇染色的选择性，进行了软骨素酶ABC和胰蛋白酶消化。硫堇染色被阻止这些酶在领土矩阵，代表interlacunar网络和软骨细胞囊。区域间基质的硫堇染色仅轻微减少，可能代表老年患者软骨中的硫酸角质素和透明质酸。硫堇染色的GMA包埋的软骨与番红O染色的石蜡包埋的切片的比较在光密度测定中显示出显著的相似性，表明硫堇与软骨中带负电荷的GAG结合的特异性。在GMA包埋软骨形态是相对完整的石蜡包埋切片相比，由于较少的收缩软骨细胞和interlacunar网络。
The usefulness of thionin for staining cartilage sections embedded in glycol methacrylate (GMA) and the effect of decalcification on cartilage sections embedded in paraffin and GMA were assessed. Short decalcification periods using 5% formic acid or 10% EDTA did not influence the staining properties or the morphology of cartilage matrix and chondrocytes. The standard stain safranin O-fast green for differential staining of cartilage was used as control in these experiments. Prolonged exposure of safranin O stained sections to fast green resulted in disappearance of the safranin O stained matrix, thereby hampering the quantitative measurement of negatively charged glycosaminoglycans (GAG). Thionin stained evenly throughout all cartilage layers, independent of the staining times. In contrast to safranin O, thionin did not show metachromasia in nondehydrated cartilage sections, which made it more suitable for assessing cartilage quality in GMA embedded cartilage. To evaluate the selectivity of thionin staining in cartilage, chondroitinase ABC and trypsin digestions were carried out. Thionin staining was prevented by these enzymes in the territorial matrix, representing the interlacunar network and the chondrocyte capsule. Staining with thionin of the interterritorial matrix was only slightly reduced, possibly representing keratan sulfate and hyaluronic acid in cartilage of elderly patients. Comparison of thionin stained GMA embedded cartilage with safranin O stained paraffin embedded sections showed significant similarity in optical densitometry, indicative of the specificity of thionin bound to negatively charged GAG in cartilage. In GMA embedded cartilage morphology was relatively intact compared to paraffin embedded sections due to less shrinkage of chondrocytes and the interlacunar network.