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Modification of methods to use Congo-red stain to simultaneously visualize amyloid plaques and tangles in human and rodent brain tissue sections.
Metabolic Brain Disease ( IF 3.6 ) Pub Date : 2020-08-27 , DOI: 10.1007/s11011-020-00608-0
Sumit Sarkar 1 , James Raymick 1 , Elvis Cuevas 1 , Hector Rosas-Hernandez 1 , Joseph Hanig 2
Affiliation  

Although there are multiple histochemical tracers available to label plaques and tangles in the brain to evaluate neuropathology in Alzheimer disease (AD), few of them are versatile in nature and compatible with immunohistochemical procedures. Congo Red (CR) is an anisotropic organic stain discovered to label amyloid beta (Aβ) plaques in the brain. Unfortunately, its use is underappreciated due to its low resolution and brightness as stated in previous studies using bright field microscopy. Here, we modified a previous method to localize both plaques and tangles in brains from humans and a transgenic rodent model of AD for fluorescence microscopic visualization. The plaque staining affinities displayed by CR were compared with fibrillar pattern labeling seen with Thioflavin S. This study summarizes the optimization of protocols in which various parameters have been finetuned. To determine the target CR potentially binds, we have performed double labeling with different antibodies against Aβ as well as phosphorylated Tau. The plaque staining affinities exhibited by CR are compared with those associated with the diffuse pattern of labeling seen with antibodies directed against different epitopes of Aβ. Neither CP13, TNT2 or TOC1 binds all the neurofibrillary tangles as revealed by CR labeling in the human brain. Additionally, we also evaluated double labeling with AT8, AT180, and PHF1. Interestingly, PHF-1 shows 40% colocalization and AT8 shows 15% colocalization with NFT. Thus, CR is a much better marker to detect AD pathologies in human and rodent brains with higher fluorescence intensity relative to other conventional fluorescence markers.



中文翻译:

修改方法以使用刚果红染色同时可视化人和啮齿动物脑组织切片中的淀粉样蛋白斑块和缠结。

尽管有多种组织化学示踪剂可用于标记大脑中的斑块和缠结以评估阿尔茨海默病 (AD) 的神经病理学,但其中很少有在性质上具有通用性并且与免疫组织化学程序兼容。刚果红 (CR) 是一种各向异性有机染色剂,被发现可标记大脑中的淀粉样蛋白 (Aβ) 斑块。不幸的是,由于其分辨率和亮度低,如先前使用明场显微镜的研究所述,它的使用被低估了。在这里,我们修改了以前的方法来定位人类大脑中的斑块和缠结,以及用于荧光显微可视化的 AD 转基因啮齿动物模型。将 CR 显示的斑块染色亲和力与用硫黄素 S 观察到的纤维状图案标记进行比较。本研究总结了对各种参数进行了微调的协议的优化。为了确定可能结合的目标 CR,我们使用针对 Aβ 和磷酸化 Tau 的不同抗体进行了双重标记。将 CR 显示的斑块染色亲和力与针对不同 Aβ 表位的抗体所见的扩散标记模式相关联的那些进行比较。CP13、TNT2 或 TOC1 都不能结合人脑中 CR 标记所揭示的所有神经原纤维缠结。此外,我们还评估了 AT8、AT180 和 PHF1 的双重标记。有趣的是,PHF-1 与 NFT 显示出 40% 的共定位,而 AT8 显示出 15% 的共定位。因此,

更新日期:2020-08-27
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