小鼠视网膜透射电镜样品制备方法与注意事项

Sample preparation and precautions for transmission electron microscopy of mouse retina

为在透射电子显微镜下能够观察到小鼠视网膜各层细胞的完整结构建立一种小鼠视网膜的电镜样品制备方法.将小鼠完整的眼球在2.5%戊二醛过夜固定,1%锇酸后固定3 h,乙醇逐级脱水,环氧树脂浸透固化,半薄定位后进行超薄切片,柠檬酸铅染色后透射电子显微镜下观察.该方法的优点在于,视网膜保护完好,电镜下能够观察到视网膜10层细胞结构,各层细胞结构完好清晰无损伤.笔者对该方法的主要注意事项进行了讨论.

In order to observe the complete structure of retinal cells in different layers of mice under transmission electron microscopy (TEM), a method for preparing electron microscopic samples of mouse retina was established. The intact eyeballs of mice were fixed in 2.5% glutaraldehyde solution overnight, then in 1% osmium acid solution for 3 hours, dehydrated in ethanol step by step, immersed and cured in epoxy resin, ultra-thin sections were made after semi-thin localization and observed under transmission electron microscopy after lead citrate staining. The advantage of this method is that the retina is well protected. Ten layers of retinal cell structure can be observed under electron microscopy. The cell structure of each layer is intact and clear without damage. The main points needing attention in this method are discussed.