Nagato Y, Mitsui T, Kushida T, Kushida H
Tokai J Exp Clin Med. 1985 Apr;10(1):37-46.
A correlative light and electron microscope method in which semi-thin sections of embedded tissue were treated with periodate acid and Schiff's reagent (PAS) was used to determine the precise localization of PAS reactive substance. Small blocks of tissue specimens were fixed with aldehyde mixtures. After dehydration, the blocks were embedded in a modified mixture of glycol methacrylate (GMA), Quetol 523, methyl methacrylate (MMA) and QCU-1. Semi-thin sections, 0.2-0.3 micron thick, were stained by the PAS reaction, followed by counterstaining with hematoxylin if necessary. It was found that PAS reaction products representing the specific sites for glycosaminoglycans and glycoproteins were seen in both light and electron microscopy. In the control experiments the specificity of the reaction was confirmed. The granules of goblet cells were well stained and contrasted by the reaction materials. The basement membrane and the microvilli of the epithelial cells appeared as the staining layer. In the spermatocytes the reaction products were demonstrated in the Golgi apparatus, acrosomal vesicles and head cap. The results indicated that the PAS deposits became electron dense when the embedding matrix had a low electron scattering property. Using this method of preparing semi-thin sections, a comparative study of the localization of glycosaminoglycans was performed.
采用一种光镜与电镜相关的方法，即用高碘酸和席夫试剂（PAS）处理包埋组织的半薄切片，以确定PAS反应性物质的精确定位。小块组织标本用醛类混合物固定。脱水后，将组织块包埋在由甲基丙烯酸乙二醇酯（GMA）、Quetol 523、甲基丙烯酸甲酯（MMA）和QCU - 1组成的改良混合物中。厚度为0.2 - 0.3微米的半薄切片通过PAS反应染色，必要时用苏木精复染。结果发现，在光镜和电镜下均可见到代表糖胺聚糖和糖蛋白特定位点的PAS反应产物。在对照实验中证实了该反应的特异性。杯状细胞的颗粒被反应物质很好地染色并形成对比。基底膜和上皮细胞的微绒毛呈现为染色层。在精母细胞中，反应产物出现在高尔基体、顶体囊泡和顶帽中。结果表明，当包埋基质具有低电子散射特性时，PAS沉积物会变得电子致密。利用这种制备半薄切片的方法，对糖胺聚糖的定位进行了比较研究。
