Histological Analysis of Mouse Brain

The mice were anesthetized with 8% chloral hydrate and then perfused with physiological saline and 4% paraformaldehyde. Brain tissues were fixed in 4% paraformaldehyde for 3 days, embedded in wax and cut into 5 µm and 10 µm slices, which were xylene dewaxed, then ethanol dehydrated. Then, sections were stained for hematoxylin and eosin (H & E) and Nissl staining (C0117, Beyotime) according to the manufacturer’s instructions. Immunohistochemistry was performed as described [17 (link)]. The primary antibodies were incubated overnight at 4 °C, and secondary antibodies were incubated for 1 h at room temperature. A Leica DM500 light microscope (Leica Microsystems, Wetzlar Germany) was used to observe the histological sections.