Histology
Histology was used to detect the expression of TRPV1 and c-fos and the successful targeting of anti-His-FVIOs to the TRPV1-expressing neurons. After AMF treatment, the mice were anesthetized and perfused transcardially with PBS and 4% paraformaldehyde (PFA) within 1.5 hours. The isolated brains were fixed in 10% PFA at 4 °C overnight and equilibrated for cryoprotection in 30% sucrose. Coronal brain slices (40 μm thick) were cut on a vibratome. The mouse anti-His tag monoclonal antibody and mouse monoclonal anti-TRPV1 antibody were used in histology. The obtained slices were stained and photographed using a laser-scanning confocal microscope. Confocal images were analyzed using ImageJ software.