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半薄切片是超薄切片前的一个重要步骤。光镜检查半薄切片,可确定电镜观察部位,判断组织固定和包埋的质量。半薄切片染色法很多,我们对几种常用方法进行了一些比较和改进,现将实践中的体会介绍如下,供参考。一、方法及结果:用Epon812包埋的组织块切成1微米或1.5微米厚的半薄切片,捞到滴有蒸馏水的载片上,在烘箱内烤干,待染色。1.甲苯胺兰染色法:将染液滴于切片上,在酒精灯上加热数秒钟,勿煮沸,流水冲洗,自然干燥,树胶封固后观察。不必经酒精及二甲苯,免切片皱缩不平。如染色适度,组织染成兰色,结构清楚,而树脂无色。2.苏木精—伊红染色法:锇酸固定、树脂包埋的组织,用此法难于着色,一般须先用氢氧化钾溶液脱去树脂,然后染色.为避免氢氧化钾液对组织可能产生的损伤,我们参考Chang法改用2.5%高碘酸处理切片,并用滴
Semi-thin section is an important step before the ultra-thin section. Light microscopy semi-thin slices, electron microscopy can determine the site to determine the quality of tissue fixation and embedding. Semi-thin slice staining a lot, we have some of the more commonly used methods to make some improvements and improvements, the experience is now introduced below, for reference. First, the method and results: The tissue block embedded with Epon812 cut into 1-micron or 1.5-micron thick semi-thin slices, fishing drip with distilled water on the slide, baking in the oven, to be dyed. 1. Toluidine blue staining: the dye drops on the slice, heated in alcohol lamp for a few seconds, do not boil, water rinse, dry naturally, observed after sealing gum. Do not have alcohol and xylene, free slices shrink wrinkles. If moderate staining, tissue stained blue, clear structure, and resin colorless. 2. Hematoxylin - eosin staining: osmium acid fixation, resin embedded in the tissue, difficult to stain with this method, the general need to first remove the resin with potassium hydroxide solution, and then staining. In order to avoid the potassium hydroxide solution on the organization Possible damage, we refer to Chang law with 2.5% periodic acid treatment section, and with drops