Kishino M, Ogawa T, Inoué T, Hamada T
J Electron Microsc (Tokyo). 1989;38(5):394-8.
Distal surface of the rat ruffle-ended ameloblasts was observed by high resolution scanning electron microscopy. Specimens fixed by perfusion with 0.5% formaldehyde and 0.5% glutaraldehyde were decalcified with ethylenediamine tetraacetic acid and freeze-fractured using dimethyl sulfoxide. They were treated with 0.1% osmium tetroxide for 96 hr to remove excess cytoplasmic matrices, dehydrated, and critical-point dried. The present method was useful for observing both surface and intracellular structures simultaneously. The dense lamina lining the distal surface of the ruffle-ended ameloblasts having been dissolved in this preparation, the surface was clearly demonstrated in three dimensions under SEM. The surface was characterized by a complex labyrinth formed by protrusion and invagination of the plasma membrane. At high magnification, two kinds of minute granules are visible: small and larger granules measured as 10-20 nm and 70 nm in diameter, respectively. The former were more numerous than the latter. Furthermore, microfibrils connecting the protrusions of the plasma membrane were observed on the distal surface. The small granules probably connect the dense lamina with the distal surface of the ameloblasts. In addition, a denuded area free from the granules was sometimes recognized on the distal surface. These surface structures of the distal end of the ameloblasts appeared to be concerned with the enamel maturation.
采用高分辨率扫描电子显微镜观察大鼠皱褶缘成釉细胞的远端面。用0.5%甲醛和0.5%戊二醛灌注固定的标本,经乙二胺四乙酸脱钙,并用二甲基亚砜进行冷冻断裂。用0.1%四氧化锇处理96小时以去除多余的细胞质基质,然后脱水并进行临界点干燥。本方法可同时观察表面结构和细胞内结构。在本制备过程中,皱褶缘成釉细胞远端面的致密板层被溶解,在扫描电子显微镜下可清晰地从三维角度观察到该表面。其表面的特征是由质膜的突出和内陷形成的复杂迷宫结构。在高倍放大下,可以看到两种微小颗粒:较小颗粒和较大颗粒的直径分别为10 - 20纳米和70纳米。前者比后者数量更多。此外,在远端面上观察到连接质膜突出部分的微原纤维。小颗粒可能将致密板层与成釉细胞的远端面连接起来。另外,在远端面上有时会识别出一个没有颗粒的裸露区域。成釉细胞远端的这些表面结构似乎与釉质成熟有关。