Fig. 5. Mouse colons colonized with human CRC patient mucosal homogenate forms mucosa-associated biofilms.

a A schematic of germ-free (GF) mouse inoculation with human biofilm-positive mucosal slurries. Mice were sacrificed 2 weeks post inoculation and processed using the Poloxamer fixative. Imaging was performed using WGA (red) for mucus, Eub (green) for bacteria, and nuclei (blue) under different conditions of inoculation into GF mice: b Human CRC biofilm isolate, and c sham. d Iterative FISH staining rounds with Bacteroidetes (green), Lachnospiraceae (red), and Gammaproteobacteria (blue), e Fusobacteria (green) probes of the tissue in b. Scale bars = 50 μm. White dotted lines represent epithelial borders. d* is a higher magnification view of red inset in d. Scale bars = 10 μm.