H&E Staining for Colon Tissue Analysis

H&E staining was conducted for the evaluation of histological changes in colon tissue according to the method described earlier [44 (link),45 (link)]. Paraffin slides were deparaffinized and rehydrated in xylene and ethanol. These slides were treated in Meyer’s hematoxylin (S3309, DAKO, Glostrup, Denmark) for 30 s and washed with water. These slides were incubated with eosin (318906, Sigma-Aldrich, St. Louis, MO, USA) for 10 s and washed with water. These slides were air-dried at room temperature and then sequentially treated in ethanol and xylene. Coverslips were covered onto the slides and mounted by Permount^® (SP15-100, Thermo Fisher Scientific, Waltham, MA, USA). Ulcerative colonic damage was evaluated with the Wallace microscopic score (Table 1) [46 (link),47 (link)].