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.