Wilcock Donna M, Gordon Marcia N, Morgan Dave
Alzheimer's Research Laboratory, School of Basic Biomedical Sciences, College of Medicine, Department of Molecular Pharmacology and Physiology, University of South Florida, Tampa, Florida 33612, USA.
Nat Protoc. 2006;1(3):1591-5. doi: 10.1038/nprot.2006.277.
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.
在本方案中，我们描述了刚果红染色方法以及一种分别定量脑组织切片中血管和实质淀粉样沉积物的方法。刚果红染色可检测淀粉样前体蛋白转基因小鼠和人类阿尔茨海默病组织的脑组织中的淀粉样沉积物。它能检测β-折叠二级结构中的致密淀粉样物质，并标记脑实质（淀粉样斑块）和血管中的淀粉样物质。血管中的嗜刚果红淀粉样物质称为脑淀粉样血管病（CAA）。迄今为止，对CAA的分析主要使用严重程度评分量表，包括定性和定量特征。在此，我们描述了一种简单的方法，可根据形态学标准对总嗜刚果红染色进行定量，并将这种染色解析为实质和血管成分。鉴于降低淀粉样蛋白疗法已进入临床试验阶段，分别定量阿尔茨海默病病理学的各种成分变得越来越重要。刚果红染色的整个过程可在通风橱中于室温（20 - 25摄氏度）下进行。染色方案包括封片时间在内需1小时30分钟。图像分析所需时间很大程度上取决于被分析样本的数量和所使用的软件。在我们的操作中，每小时可收集30张图像，并在另外2小时内进行定量分析。
