U mwig3 filter set

Immunohistochemical staining was performed according to the procedure described in previous reports [21 (link), 31 (link)–33 ]. Briefly, one hour after the post-OIT non-heated OVA challenge, the proximal colon was excised, fixed by immersion in 4% paraformaldehyde and then embedded in OCT compound. Cutted sections (30 μm) were exposed to antiserum against mMCP-1, a marker of mouse mucosal mast cells (1:5,000; Moredun Scientific, Scotland, UK) and then incubated with Cy3-conjugated sheep anti-donkey IgG (1:200; Jackson Immunoresearch Laboratories). The immunostained sections were examined using a fluorescence microscope (IX71 system; Olympus, Tokyo, Japan) with a U-MWIG3 filter set (Olympus) and photographed using an Olympus digital camera (DP70; Olympus). The histological staining was quantitatively analyzed using ImageJ software.

The colonic sections were treated with a rat IgG anti-mouse CD11c antibody, rat IgG anti-mouse CD4 antibody, rabbit IgG anti-rat CGRP antibody, or goat IgG anti-mouse CCL21 antibody for 12–18 h at 4 °C. After the incubation, the sections were incubated with a suitable secondary antibody for 2 h at RT. The sections were observed using a confocal laser scanning microscope (LSM700; Carl Zeiss Japan, Tokyo, Japan) or a fluorescence microscope (IX71 system; Olympus, Tokyo, Japan) with a U-MWIG3 filter set (Olympus) and photographed using an Olympus digital camera (DP70; Olympus).