Shea S M
J Cell Biol. 1971 Dec;51(3):611-20. doi: 10.1083/jcb.51.3.611.
Among the techniques which have been reported to stain the surface coat of cells, for electron microscopy, is lanthanum staining en bloc. Similarly, the presence of the cationic dye, Alcian blue 8GX, in a primary glutaraldehyde fixative has been reported to improve the preservation of the surface coat of cells of many types; however, the preserved coat is not very electron opaque unless thin sections are counterstained. The present paper shows that for several rat tissues lanthanum staining en bloc is an effective electron stain for the cell surface, giving excellent contrast, if combined sequentially with prefixation in an aldehyde fixative containing Alcian blue. The cationic substance cetylpyridinium chloride was found to have a similar effect to that of Alcian blue in enhancing the lanthanum staining of the surface coat material of the brush border of intestinal epithelial cells. The patterns of lanthanum staining obtained for the tissues studied strikingly resemble those reported in the literature where tissues are stained by several standard methods for demonstrating mucosubstances at the ultrastructural level. This fact and the reproduction of the effect of Alcian blue by cetylpyridinium chloride constitute a persuasive empirical argument that the material visualized is a mucopolysaccharide or mucopolysaccharide-protein complex.
在已报道的用于电子显微镜观察细胞表面被覆层染色的技术中,有整体镧染色法。同样,据报道,在初次戊二醛固定剂中加入阳离子染料阿尔辛蓝8GX可改善多种类型细胞表面被覆层的保存效果;然而,除非对薄切片进行复染,否则保存的被覆层在电子显微镜下的不透明度并不高。本文表明,对于几种大鼠组织,整体镧染色法是一种有效的细胞表面电子染色方法,如果在含有阿尔辛蓝的醛类固定剂中进行预固定后再依次进行镧染色,可产生极佳的对比度。发现阳离子物质十六烷基氯化吡啶在增强肠道上皮细胞刷状缘表面被覆层物质的镧染色方面与阿尔辛蓝具有相似的效果。所研究组织的镧染色模式与文献中报道的通过几种超微结构水平显示粘多糖物质的标准方法对组织进行染色的模式极为相似。这一事实以及十六烷基氯化吡啶对阿尔辛蓝效果的重现构成了一个有说服力的经验论据,即所观察到的物质是一种粘多糖或粘多糖 - 蛋白质复合物。