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免疫胶体在光学显微镜中的应用。II. 利用凝集素-金或糖蛋白-金复合物在石蜡切片中显示凝集素结合位点。

Applications of immunocolloids in light microscopy. II. Demonstration of lectin-binding sites in paraffin sections by the use of lectin-gold or glycoprotein-gold complexes.

作者信息

Roth J

出版信息

J Histochem Cytochem. 1983 Apr;31(4):547-52. doi: 10.1177/31.4.6186737.

Abstract

A new procedure is presented for the light microscopic demonstration of specific sugar sequences of oligosaccharides in glycoconjugates by lectins combined with the colloidal gold marker system. Tissue sections from aldehyde-fixed and paraffin embedded rat kidney were stained either in a one-step method with lectin directly bound to particles of colloidal gold or in a two-step method using non-labeled lectin and glycoprotein labeled with colloidal gold. In both methods the presence of lectin-binding sites in the tissue sections is revealed by the appearance of a red coloration that is due to the accumulation of gold particles. The high specificity of the technique is combined with a good sensitivity and resolution as demonstrated by a differential plasma membrane staining in renal epithelial cells. The lectin-gold or glycoprotein-gold complexes remain stable for months and produce a permanent nonbleaching staining.

摘要

本文介绍了一种新方法,通过凝集素结合胶体金标记系统,在光学显微镜下显示糖缀合物中寡糖的特定糖序列。取自醛固定、石蜡包埋大鼠肾脏的组织切片,可用一步法直接用与胶体金颗粒结合的凝集素染色,也可用两步法,即使用未标记的凝集素和用胶体金标记的糖蛋白染色。在这两种方法中,组织切片中凝集素结合位点的存在都通过金颗粒的积累而呈现红色来显示。如肾上皮细胞中质膜的差异染色所示,该技术具有高特异性,同时具有良好的敏感性和分辨率。凝集素-金或糖蛋白-金复合物可保持稳定数月,并产生永久性不褪色染色。

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