Ix83p2zf microscope

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).

The frozen tissue sections (6 μm) were dried at room temperature for 60 min and rinsed with PBS two times. The slides were then rinsed in ddH₂O for 1 min, dipped into Hemaroxalinstain for 3 min, rinsed with ddH₂O twice, dipped fast 8–12 times in acid ethanol, rinsed 2 × 1′ tap water, rinsed 1 × 2′ deionized water, 1 × 30 seconds Eosin, dipped in 90%, 95% and 100% ethanol for 3 min each, then dipped in 100% ethanol again for 3 min, and dipped in two changes of 100% xylene for 3 min each. The slides were then mounted with DePex mounting medium (VWR 361254D), and images were taken with an Olympus (IX83P2ZF) microscope.