Sun Anyang, Nguyen Xuan V, Bing Guoying
Department of Anatomy & Neurobiology, University of Kentucky Medical Center, Lexington, KY 40536-0098, USA.
J Histochem Cytochem. 2002 Apr;50(4):463-72. doi: 10.1177/002215540205000403.
An improved thioflavin-S stain, Gallyas silver stain, and two immunostainings were quantitatively compared for demonstration of neurofibrillary tangles (NFTs) on the same sections. Sections of hippocampal formation from seven cases of Alzheimer's disease (AD) were immunofluorescently stained with a commercially available polyclonal NFT antibody or a PHF-1 monoclonal antibody, followed by an improved thioflavin-S stain, and finally by Gallyas silver staining. The thioflavin-S method was improved by using a combination quenching method that removes background autofluorescence without remarkable tissue damage and by post-treatment with concentrated phosphate buffer, which minimizes photobleaching. PHF-1 or NFT immunostaining is much less sensitive than the improved thioflavin-S staining and Gallyas silver staining, particularly in the transentorhinal region. Moreover PHF-1 immunoreactivity varied greatly among AD individuals. Thioflavin-S staining and Gallyas silver staining show almost the same sensitivity in NFT demonstration, but only the former depends on the secondary protein structure of NFTs. This study suggests that the improved thioflavin-S staining is a simple, sensitive, and consistent method for demonstration of neurofibrillary pathology.
对改良硫黄素 - S染色、加利亚斯银染色以及两种免疫染色进行了定量比较，以在同一切片上显示神经原纤维缠结（NFTs）。对7例阿尔茨海默病（AD）患者的海马结构切片先用市售多克隆NFT抗体或PHF - 1单克隆抗体进行免疫荧光染色，接着进行改良硫黄素 - S染色，最后进行加利亚斯银染色。硫黄素 - S方法通过采用一种组合淬灭法进行了改良，该方法可去除背景自发荧光且不会对组织造成明显损伤，还通过用浓缩磷酸盐缓冲液进行后处理，从而将光漂白降至最低。PHF - 1或NFT免疫染色的敏感性远低于改良硫黄素 - S染色和加利亚斯银染色，尤其是在内嗅区。此外，AD个体之间的PHF - 1免疫反应性差异很大。硫黄素 - S染色和加利亚斯银染色在显示NFT方面的敏感性几乎相同，但只有前者依赖于NFT的二级蛋白质结构。本研究表明，改良硫黄素 - S染色是一种用于显示神经原纤维病理的简单、敏感且一致的方法。
