Distal colon segments in the same defined region were collected carefully and were transferred immediately to 10% neutral-buffered formalin. After paraffin embedding, colon fragments were generated into 5 μm cross-sections. The sections were stained with hematoxylin and eosin (H&E) and then scanned by a NanoZoomer Digital Pathology system (Hamamatsu Photonics, KK, Japan) for further histopathological analysis. At least three slides were randomly selected and observed by a blinded pathologist. The histological activity index (HAI) is an established scoring system based on inflammatory cell infiltration (1–3), both in terms of severity and extent, and intestinal architecture (1–3), which includes epithelial changes and mucosal architecture, as previously described (Erben et al., 2014 (link)).
The colon sections were stained with the intestinal barrier markers ZO-1 and Occludin (both Proteintech, Rosemont, IL, United States) with a previously described standard immunofluorescence analysis process (Chung et al., 2014 (link)). Images were captured and visualized using a Zeiss LSM T-PMT confocal microscope (Zeiss, Jena, Germany).
The colon sections were stained with the intestinal barrier markers ZO-1 and Occludin (both Proteintech, Rosemont, IL, United States) with a previously described standard immunofluorescence analysis process (Chung et al., 2014 (link)). Images were captured and visualized using a Zeiss LSM T-PMT confocal microscope (Zeiss, Jena, Germany).
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