Histological Analysis of Colon Injury

One centimeter of the distal colon was fixed in 10% formaldehyde followed by paraffin embedding. The colon tissue was then sectioned into 5μm thick sections dewaxed in xylene, dehydrated with gradient ethanol, and stained with hematoxylin for five minutes and with eosin for one minute. The colon sections were then observed under a microscope (Olympus, BX53, Japan). The villus length (μm) of the HE-stained sections was determined using Case Viewer 2.0 software (3DHISTECH Ltd.). Villus length was defined as the distance from the top of the villi to the muscularis of the intestinal wall. The gross morphological score for colon injury was evaluated based on the parameters shown in Table 2 (1 (link)). The samples were evaluated by experienced pathologists who were blinded to each other.