Staining and Quantification of β-Amyloid Pathology in Transgenic Mouse Models of Alzheimer's Disease - PubMed (original) (raw)

Staining and Quantification of β-Amyloid Pathology in Transgenic Mouse Models of Alzheimer's Disease

Amy Christensen et al. Methods Mol Biol. 2020.

Abstract

Studies of Alzheimer's disease (AD) using experimental systems most often involve transgenic mouse models that are characterized by neural accumulation of β-amyloid protein (Aβ), which is widely hypothesized to have a key role in AD pathogenesis. Quantification of Aβ in transgenic mice typically is accomplished through both biochemical and histochemical approaches. In this chapter, we describe two techniques for the histological detection of Aβ, immunostaining with Aβ antibodies and staining with the amyloid dye thioflavin S, and its quantification using digital imaging.

Keywords: Immunohistochemistry; Quantitative imaging; Thioflavin S; Transgenic mice; β-Amyloid.

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Figures

Fig. 1

Fig. 1

Illustration of image collection and region of interest determination. Horizontal section of EFAD transgenic mouse showing extensive pathology by amyloid β immunoreactivity using protocol described in Subheading 3.1. Boxes show pattern of image collection using 20 × objective for subiculum (Sub) and CA1 hippocampus (CA1). Inset shows larger view of first subiculum image. Using the polygon function, the region of interest (ROI) is defined (dotted red line) to include subiculum but not adjacent cortex (Ctx) and white matter (WM) (as described in Subheading 3.4)

Fig. 2

Fig. 2

Examples of image transformation prior to quantification. Panels show a single microscopic field from subiculum in an EFAD transgenic mouse of either amyloid β immunoreactivity (left column) or thioflavin S staining (right column) at initial image collection of labeling (Captured image), following conversion to grayscale (Grayscaled image), and after the threshold step (Thresholded image). The thresholded images are suitable for quantification (described in Subheadings 3.4 and 3.5). Note that the ROI is not shown

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