We performed ex vivo tissue imaging of intestinal bacterial GUS activity through quantification of fluorescence following hydrolysis of FDGlcU (Sigma-Aldrich) to fluorescein. Three hours prior to imaging, each animal was gavaged with FDGlcU (7.3 μmol/kg). Immediately before imaging, mice were euthanized, and abdominal organs were excised and imaged using an In Vivo Xtreme 4MP imaging platform (Bruker). Reflectance imaging (2-s exposure time) was followed by fluorescence imaging (5-s exposure time, 470-nm excitation and 535-nm emission filters) and x-ray imaging (10-s exposure time). Pixel binning was kept constant at 4 × 4. Images were acquired and analyzed using the Bruker molecular imaging software MI SE v7.1.3.20550. GUS activity was quantified by measuring the mean fluorescence after background subtraction in a given region of interest, which was kept constant over the time period of imaging.

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