Computational protocol: Obesity Accelerates Alzheimer-Related Pathology in APOE4 but not APOE3 Mice

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Protocol publication

[…] Fixed hemi-brains were fully sectioned in the horizontal plane at 40 μm using a vibratome (Leica Biosystems). Every eighth section was stained for Thio-S (#230456, Sigma-Aldrich) using standard methodology. Sections were mounted and allowed to dry overnight, after which they were washed three times in 50% ethanol for 5 min each, then washed in double-distilled H2O before being incubated for 10 min in 1% Thio-S dissolved in H2O. Stained slides were then rinsed in 70% ethanol before being dehydrated and coverslipped in aqueous anti-fade mounting medium (Vector Laboratories). Digital images were captured at 20× magnification using an Olympus BX50 microscope equipped with a DP74 camera and CellSens software (Olympus). The number of spherical thioflavin-positive deposits were counted using NIH ImageJ 1.50i (United States National Institutes of Health) with the cell counter plugin to mark stained plaque-like structures. Thioflavin-positive deposits were counted in entorhinal cortex (three fields/section), subiculum (two fields/section), and hippocampal subfields CA1 (three fields/section) and CA2/3 (three fields/section), across four sections per animal, for a total of ∼44 fields per brain. […]

Pipeline specifications

Software tools cellSens, ImageJ
Application Microscopic phenotype analysis
Organisms Mus musculus, Homo sapiens, Rattus norvegicus
Diseases Alzheimer Disease
Chemicals Carbohydrates