Analysis of the mucus layer and goblet cells was made as previously described.89 (link) Briefly, paraffin sections of 5 μm were stained with alcian blue. Images were captured at ×20 magnification and obtained using a SNC400 slide scanner and digital Image Hub software 561 (Leica Biosystems, Wetzlar, Germany). Analyses were performed using ImageJ (version 1.48 r, National Institutes of Health, Bethesda, Maryland, USA) in a blinded manner. For the mucus layer thickness, two to four fields were used for each mouse and a minimum of 50 different measurements (up to 200) were made perpendicular to the inner mucus layer per field. Each value represents the mean of the different measurements per field. For the goblet cells, the luminal side, muscularis mucosae, submucosa, and muscle layer were removed using ImageJ software and the blue area and the total area were measured separately in the remaining mucosal part of the colon. The proportion of the goblet cells was quantified based on the ratio between the blue area over the total area.
Histological Analysis of Adipose Tissue and Colonic Mucus
Analysis of the mucus layer and goblet cells was made as previously described.89 (link) Briefly, paraffin sections of 5 μm were stained with alcian blue. Images were captured at ×20 magnification and obtained using a SNC400 slide scanner and digital Image Hub software 561 (Leica Biosystems, Wetzlar, Germany). Analyses were performed using ImageJ (version 1.48 r, National Institutes of Health, Bethesda, Maryland, USA) in a blinded manner. For the mucus layer thickness, two to four fields were used for each mouse and a minimum of 50 different measurements (up to 200) were made perpendicular to the inner mucus layer per field. Each value represents the mean of the different measurements per field. For the goblet cells, the luminal side, muscularis mucosae, submucosa, and muscle layer were removed using ImageJ software and the blue area and the total area were measured separately in the remaining mucosal part of the colon. The proportion of the goblet cells was quantified based on the ratio between the blue area over the total area.
Corresponding Organization : Walloon Excellence in Lifesciences and Biotechnology
Other organizations : University of Gothenburg
Variable analysis
- None specified
- Adipocyte diameter
- F4/80 positive areas in the adipose tissue
- Mucus layer thickness
- Goblet cell proportion
- Subcutaneous adipose tissue samples
- Paraffin sections of 5 μm
- Staining with hematoxylin and eosin, alcian blue
- Image capturing using SCN400 slide scanner and digital Image Hub software
- Analysis using ImageJ software
- Positive control: Not specified
- Negative control: Not specified
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