从半薄切片上去除Epon树脂的简单方法。亲和素生物素技术的应用。

The Histochemical Journal Pub Date : 1995-03-01
S Vidal, M Lombardero, P Sánchez, A Román, L Moya
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引用次数: 0

摘要

描述了一种从半薄的1微米切片中去除Epon树脂的简单方法,这种方法可以进行良好的包埋后免疫组织化学染色(亲和素-生物素复合物技术)。建立了以2%多聚甲醛和0.5%戊二醛固定于0.1 M钙酸钠缓冲液pH 7.4-7.6中牛垂体腺垂体生长激素和催乳素的检测方法。结果表明,在应用免疫组织化学试剂之前去除环氧包埋介质对于成功定位两种激素的抗原决定因素至关重要。只有在用氢氧化钾溶液(无水甲醇和环氧丙烷的混合物)处理切片后,才能获得免疫细胞化学反应性。在此处理之后,用4%过氧化氢或饱和的超碘酸钠水溶液进行额外处理,获得了增强的免疫反应性。由于Epon去除液制备简单,结构保存良好且不破坏抗原决定因子,麦克斯韦溶液被认为是一种很好的蚀刻剂,可用于半薄切片的免疫组织化学研究,并取得了很好的结果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
An easy method for the removal of Epon resin from semi-thin sections. Application of the avidin-biotin technique.

A simple procedure is described for removing Epon resin from semi-thin 1 micron sections, which permits excellent postembedding immunohistochemical staining (avidin-biotin complex technique). The procedure was developed for the detection of growth hormone and prolactin in bovine adenohypophysis fixed with 2% paraformaldehyde and 0.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.4-7.6. The results indicate that the removal of the epoxy embedding medium prior to the application of the immunohistochemical reagents was essential for the successful localization of the antigenic determinants of the two hormones. The immunocytochemical reactivity was obtained only after treating the sections with a solution of potassium hydroxide in a mixture of absolute methyl alcohol and propylene oxide (Maxwell's solution). An enhanced immunoreactivity was obtained when this treatment was followed by an additional treatment with either 4% hydrogen peroxide or a saturated aqueous solution of sodium metaperiodate. Because of the easy preparation of the Epon removal solution and the good structural preservation without damage to the antigenic determinants, Maxwell's solution is suggested as a good etching agent which can be used in immunohistochemical studies on semi-thin sections with excellent results.

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