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Catalog Item
Ex vivo assessment of barrier function/gut permeability (minimum of 8 samples)
Common Name(s): Gut Microbiome (Microbiota)

CenterUniversity of California Davis
Center CoreP&B Core
Test No.D4301
Test PricePer sample fee (minimum of 8 samples): $7/$10 (Internal University of California Fee / All Other Institutions)
DescriptionGut tissue (by region- e.g ileum & colon) will be opened along the mesenteric
border and mounted in Ussing chambers (Physiologic Instruments, San Diego, CA,
USA), exposing 0.3 cm2 of tissue surface area to 2.5ml of oxygenated
Krebs-glucose (10mM) and Krebs-mannitol (10mM) at 37°C on the serosal and
luminal sides, respectively. The paracellular pathway and transcellular pathway
will be measured as the flux of FITC-Dextran 4000 (FD-4, Sigma –Aldrich) and
horseradish peroxidase (HRP Type VI, Sigma Aldrich), respectively. FD-4
(400µg/ml) and HRP (200µg/ml) will be added to the mucosal chamber and samples
will be collected from the serosal chamber every 30 min for 2 hours.
Concentration of FD-4 is measured via fluorescence at excitation 485 nm,
emission 538 nm. O-dianisidine substrate is used to detect HRP at absorbance 450
nm.
DataData will be collected for this catalog item
Keywordsbarrier function, gastrointestinal tract, GI function, gut, intestinal
permeability, intestine, microbiome
Tissuescaecum, colon, duodenum, ileum, intestine, intestine epithelium, jejunum



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