Quantification of cerebral amyloid angiopathy and parenchymal amyloid plaques with Congo red histochemical stain

• Published in: Nature protocols Vol. 1; no. 3; pp. 1591 - 1595
• Main Authors: Morgan, Dave, Wilcock, Donna M, Gordon, Marcia N
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01.11.2006
• Subjects: Alzheimer Disease - metabolism; Alzheimer's disease; Amyloid beta-protein; Amyloid beta-Protein Precursor - genetics; Animals; Blood vessels; Brain; Brain - metabolism; Brain research; Cerebral Amyloid Angiopathy - metabolism; Clinical trials; Congo Red; Health aspects; Histochemistry; Histocytochemistry - methods; Humans; Hydrogen bonds; Image Processing, Computer-Assisted; Measurement; Methods; Mice; Mice, Transgenic; Pathology; Peptides; Plaque, Amyloid - metabolism; Proteins; Software; Stains & staining; Stains and staining (Microscopy); Tissues; United States
• ISSN: 1754-2189; 1750-2799
• 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.