Thioflavin-S Staining of Amyloid-Beta Aggregation in Cerebral Hypoperfused Mouse Brain Tissue

Take a brain tissue section from a mouse subjected to mild chronic cerebral hypoperfusion, or MCCH, and injected with amyloid-beta peptides.

MCCH is characterized by reduced blood flow to the brain. This limits oxygen and nutrient supply to the endothelial cell layer of cerebral blood vessels, damaging the cells.

The damage increases the vascular wall permeability, allowing the injected amyloid-beta peptides to accumulate in the wall and form plaques.

Wash the section with a buffer to remove residual chemicals.

Incubate with thioflavin S, a fluorescent dye that binds to the amyloid plaques.

Wash the stained section with decreasing alcohol concentrations to remove any unbound dye and rehydrate the tissue.

Place the section in the buffer to maintain hydration.

Mount the section using a mounting medium that preserves the dye's fluorescence intensity.

Under a fluorescence microscope, examine the presence of fluorescently labeled amyloid deposits to confirm MCCH-induced amyloid-beta aggregation.