In the current protocol, we describe the Congo red staining method and a method for separately quantifying vascular and parenchymal amyloid deposits in brain tissue sections. Congo red staining detects amyloid deposits in brain tissue of amyloid precursor protein transgenic mice and human Alzheimer's tissue. It detects compacted amyloid in a beta-sheet secondary structure and labels amyloid in both the brain parenchyma (amyloid plaques) and blood vessels. Congophilic amyloid in blood vessels is called cerebral amyloid angiopathy (CAA). To date, analysis of CAA has largely used a severity rating scale, including both qualitative and quantitative characteristics. Here, we describe a simple method for quantifying total Congophilic staining and resolution of this staining into the parenchymal and vascular components based on morphological criteria. It is becoming increasingly important to separately quantify various components of the Alzheimer's pathology, given the advancement of amyloid-lowering therapies into clinical trials. The entire procedure for the Congo red staining can be performed at room temperature (20-25 degrees C) in a fume hood. The staining protocol should take 1 h 30 min including time for coverslipping slides. Time required for image analysis depends greatly on the number of samples being analyzed and the software being used. In our hands, 30 images can be collected per hour and quantified in a further 2 h.

译文

在当前方案中,我们描述了刚果红染色方法以及一种用于分别定量脑组织切片中血管和实质淀粉样蛋白沉积物的方法。刚果红染色检测淀粉样蛋白前体蛋白转基因小鼠和人类阿尔茨海默氏症组织的脑组织中的淀粉样蛋白沉积。它在 β-sheet二级结构中检测压实的淀粉样蛋白,并在脑实质 (淀粉样斑块) 和血管中标记淀粉样蛋白。血管中的嗜血淀粉样蛋白被称为脑淀粉样血管病 (CAA)。迄今为止,对CAA的分析主要使用了严重性等级量表,包括定性和定量特征。在这里,我们描述了一种基于形态学标准的定量总嗜血染色和将该染色拆分为实质和血管成分的简单方法。鉴于降低淀粉样蛋白疗法已进入临床试验,单独量化阿尔茨海默氏病病理学的各个组成部分变得越来越重要。刚果红染色的整个过程可以在室温 (20-25 ℃) 在通风橱中进行。染色方案应花费1小时30分钟,包括盖玻片的时间。图像分析所需的时间在很大程度上取决于所分析的样本数量和所使用的软件。在我们的手中,每小时可以收集30张图像,并在2小时内进行量化。

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