Morphometric Analysis of Intestinal Tissues

As previously described [26 (link)], the segments of the jejunum and ileum tissues were fixed in 4% formaldehyde and embedded in paraffin. Sections of 5 μm thickness were stained with hematoxylin and eosin (H&E). Photomicrographs were acquired using an Olympus IX83P2ZF microscope (Olympus, Tokyo, Japan). The morphometric analysis was performed on 10 randomly-selected, well-oriented villi and crypts per piglet. A computerized microscope-based image analyzer (Olympus dotslide Virtual Slide System, Tokyo, Japan) was used to determine the height of villus (from the tip of the villus to the villus-crypt junction) and crypt depth (from the crypt-villus junction to the base of the crypt).