Kao Y C, Lichtenberger L M
Department of Physiology and Cell Biology, University of Texas Medical School, Houston 77225.
J Histochem Cytochem. 1990 Mar;38(3):427-31. doi: 10.1177/38.3.1689341.
Previous results from our laboratory employing the phospholipid-selective cytochemical stain iodoplatinate (IP) suggest that surfactant-like phospholipids (SLPL) are intracellularly contained within rodent gastric mucous cells and are occasionally seen extracellularly within the mucous gel layer. This hydrophobic lipid coating may provide the stomach with a protective water-repellent lining against the corrosive gastric juice in the lumen. Extracellular SLPL are frequently removed during tissue processing for electron microscopy. In this study, we developed a simple method employing an agarose-embedding technique to retain these extracellular SLPL in gastric mucosa excised from rats pre-treated with prostaglandin (to stimulate gastric surfactant/mucus secretion). With the help of polypropylene supporting screens and cassette carriers, thin slices of agarose-embedded gastric mucosa were well preserved and uniformly stained with IP. Extracellular myelin figures were well retained over the interfoveolar surface as well as in the pit region. The IP-reactive substances were seen within or coating the surface of the mucous gel. Our results also indicate that agarose is useful not only for supporting soft tissue while preparing sections with a microslicer but also for preservation of extracellular lipoidal material for electron microscopic observation.
我们实验室先前使用磷脂选择性细胞化学染色碘铂酸盐(IP)的研究结果表明,表面活性剂样磷脂(SLPL)在啮齿动物胃黏液细胞内有细胞内储存,偶尔也可见于黏液凝胶层的细胞外。这种疏水性脂质涂层可为胃提供一层防水衬里,以抵御管腔内腐蚀性胃液的侵蚀。在用于电子显微镜检查的组织处理过程中,细胞外的SLPL常常会被去除。在本研究中,我们开发了一种简单的方法,采用琼脂糖包埋技术来保留这些细胞外SLPL于从用前列腺素预处理(以刺激胃表面活性剂/黏液分泌)的大鼠切除的胃黏膜中。借助聚丙烯支撑筛网和标本盒载体,琼脂糖包埋的胃黏膜薄片保存良好,并用IP均匀染色。细胞外髓鞘样结构在小窝间表面以及隐窝区域均得到良好保留。在黏液凝胶内部或其表面可见IP反应性物质。我们的结果还表明,琼脂糖不仅在使用切片机制作切片时对支撑软组织有用,而且对于保存用于电子显微镜观察的细胞外脂质物质也很有用。