Computational protocol: Delivery of a mucin domain enriched in cysteine residues strengthens the intestinal mucous barrier

Similar protocols

Protocol publication

[…] Fluorescent beads with a diameter of 1 μm (Biovalley, Marne La Vallee, France) were loaded (10 μL; ~106 beads) at the surface of fresh colon samples that were opened lengthwise (12 WT and 11 Tg mice, respectively). After 45 min, particle penetration was recorded by confocal imaging in an XY stack using a Zeiss LSM 710 confocal microscope and the ZEN 2009 software (Zeiss, Germany). Images were analyzed using the ImageJ software (National Institutes of Health, Bethesda, MD). For each sample tissue, the total number of beads was normalized to 1,000 and the number of sections was normalized to 210. The particle distribution is presented as the percentage of beads/section. [...] Multilabel immunofluorescence analysis was performed on a Leica DM4000B or a Zeiss LSM 710 confocal microscope. Images were acquired and minimally processed by importing them into the GNU image manipulation program GIMP and the ImageJ software. Quantification of Muc2 and UEA1 or SNA colocalization in the ileum and in the colon (n = 7 mice/group) was performed on 4–7 villi or crypts/mouse. Fluorescence was measured and expressed as the UEA1/Muc2 and SNA/Muc2 (data not shown) ratio relative to the area of the villus or the crypt. […]

Pipeline specifications

Software tools ImageJ, GIMP
Applications Miscellaneous, Laser scanning microscopy, Microscopic phenotype analysis
Organisms Mus musculus, Escherichia coli, Citrobacter rodentium
Diseases Colitis, Inflammatory Bowel Diseases
Chemicals Cysteine