Each tissue section was incubated in 500 μM of thioflavin-S (Sigma-Aldrich) solution, dissolved in 50% ethanol, for 7 min at room temperature, as described elsewhere [33 (link)]. Hoechst-33342 (10μg/mL, Sigma-Aldrich) was used to observe the nuclear morphology. Images were captured and analyzed in AXIO Observer inverted microscope (Carl Zeiss). The number and area of plaques detected by thioflavin-S were quantified using ImageJ software.
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