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Updated: Sep 28, 2026

Three-dimensional Quantification of Intestinal Mucus Using Whole-mount Tissue Imaging
Published on: September 12, 2025
An improved technique for the histological evaluation of the mucus-secreting status in rat cecum
Takamitsu Tsukahara1, Yoshie Iwasaki, Keizo Nakayama
1Laboratory of Animal Science, Kyoto Prefectural University, Shimogamo, Kyoto 606-8522, Japan.
Abstract:
Mucin secreted into the alimentary tract often forms a mucus layer on the mucosa and is believed to protect the underlying epithelium against various factors in the lumen. We developed an improved histological technique for the evaluation of the mucus layer in the rat cecum. We used this technique to compare the effect of three nonstarch poly- and oligosaccharides on the status of mucus layer. Rats were divided into four groups (fiber-free [FF], cellulose [CEL], fructooligosaccharide [FOS], or guar gum [GG]). The frozen cecum with its contents was cut into cross-sections (5 mm thick) and fixed overnight in half-strength Bouin's solution. The sections were then transferred to 80% ethanol for 24 h. After being stained with alcian green, the mucus layers were clearly visualized in thin sections of the rat cecum, except for those that received FOS where the mucus layer had disappeared; the strong signal of mucus was seen in the cecal digesta of FOS-fed rats. Our histological method successfully provided information about the status of mucus layer that is important for an assessment of the epithelial state in the intestine.
