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Frozen hippocampal tissues were cut into 30 μm sections using a cryostat microtome (Leica CM1850; Leica Microsystems). The resulting pieces of tissue were thoroughly washed with distilled water for 5 min, and then transferred to gelatin-coated slides and placed in 1% Thioflavine T for 5 min, followed by ethanol dehydration (50%, 70%, 90%, and 100%, 2 min in each grade). The dehydrated samples were then mounted with mounting medium (Fluoromount, Sigma). Thioflavin T staining was examined by using a fluorescence microscope.
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