Histological Analysis of Intestinal Mucosal Changes

Paraffin-embedded sections were deparaffinised using Clear-Advantage Xylene Substitute (Polyscience, USA) and rehydrated through an ethanol gradient. Haematoxylin and Eosin (H&E) staining for epithelium integrity was performed immediately after deparaffinisation by a Haematoxylin and Eosin Stain Kit (Vector Laboratories, USA) following the manufacturer’s instructions. In addition, replicate slide sections were stained with Alcian Blue/Periodic acid–Schiff (PAS) staining following the manufacturer’s instructions (Polyscience, USA) to visualise mucosal variations. Slides were then imaged using an Olympus BX51 upright microscope and sections were scored as described in previous work [42 (link)–45 (link)]. All scoring was blinded to experimental conditions with reference to the group only fed the prebiotic diet (no bifidobacterial gavage) to evaluate gross changes compared to control samples in (1) gland number (colon) or villus structure (small intestine), (2) oedema and at the cellular level with (3) goblet cell number, (4) mucus thickness, and (5) acid/neutral mucin distribution from crypts to lumen.

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Hoedt E.C., Hueston C.M., Cash N., Bongers R.S., Keane J.M., van Limpt K., Ben Amor K., Knol J., MacSharry J, & van Sinderen D. (2023). A synbiotic mixture of selected oligosaccharides and bifidobacteria assists murine gut microbiota restoration following antibiotic challenge. Microbiome, 11, 168.

Publication 2023

BX51 upright microscope

Acid Alcian blue Bifidobacterial Cellular Colon Crypts Diet Eosin Epithelium Ethanol Gavage Haematoxylin stain Microscope Mucin Mucosal Mucus Oedema Paraffin Periodic acid Prebiotic Replicate Small intestine Vector Xylene

Corresponding Organization :

Other organizations : APC Microbiome Institute, University College Cork, Nutricia Research (Netherlands), Danone (Netherlands)

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