Instituto de Neurociencias del Principado de Asturias, Dpto. Morfología y Biología Celular, Facultad de Biología y Medicina, Universidad de Oviedo, Oviedo, Spain.
J Alzheimers Dis. 2013;35(3):589-97. doi: 10.3233/JAD-122386.
A highly selective, rapid, inexpensive, simple, and immunocytochemical compatible fluorescence staining method for Alzheimer's disease hallmark lesions applicable to sections of human specimens embedded in paraffin is described. Human necropsy material was fixed in buffered formalin, sectioned at 10 μm, mounted on slides, deparaffinized, and partially hydrated (70% ethanol). After partial hydration, sections were stained for 10 min in a solution of 0.2% Congo red in 70% isopropanol. After washing in 70% isopropanol and rehydration, auto-fluorescence of sections were quenched (optional) and processed for immunocytochemistry (optional). Finally, sections were mounted in an adequate mounting medium. Amyloid deposits appear pink at light microscopy and all Alzheimer's disease hallmark lesions appear orange or red under fluorescence microscopy using blue or green exciting light, respectively. The present method can be used in combination with all pre- or post-immunocytochemical techniques.
本文描述了一种适用于石蜡包埋人标本切片的、针对阿尔茨海默病标志性病变的高度选择性、快速、廉价、简单且与免疫细胞化学兼容的荧光染色方法。人尸检材料用缓冲福尔马林固定，切成 10μm 厚的切片，贴在载玻片上，脱蜡并部分水化（70%乙醇）。部分水化后，将切片在 70%异丙醇中 0.2%刚果红溶液中染色 10 分钟。在 70%异丙醇中洗涤并再水化后，（可选）淬灭切片的自发荧光并进行免疫细胞化学处理（可选）。最后，将切片用适当的封固介质封固。在光镜下，淀粉样沉积物呈粉红色，在使用蓝色或绿色激发光的荧光显微镜下，所有阿尔茨海默病标志性病变分别呈现橙色或红色。本方法可与所有免疫细胞化学前或后技术结合使用。
