Coolscope light microscope

Topiramate-treated and control mice were sacrificed after 21 days of treatment. They were perfused intracardially with 4 °C, 4% paraformaldehyde in PBS (phosphate buffer saline). Brains were quickly removed and post-fixed in 4% paraformaldehyde overnight at 4 °C. Brains were then cut into two hemispheres, embedded in paraffin, serially sectioned (3 µm), and mounted on silan-covered slides. Hemispheres sections were stained by immunohistochemistry as described previously, with the following antibodies: β-amyloid (1:100; Abcam, Cambridge, UK) for Aβ deposition, and Iba-1 (1:200; Wako, Neuss, Germany) for activated microglia. For data analysis, hemisphere sections were examined using a Nikon Coolscope light microscope. Two independent observers randomly numbered and analyzed the different sections. The total number of Aβ plaques and microglia cells positive for Iba-1 staining in the neocortex and hippocampus was calculated. Images of cross-sections were captured and areas of interest (neocortex and hippocampus) were outlined and analyzed. Furthermore, area percentages of specific immunoreactivity (IR) were also calculated.