Changing the level of mucin within ASM+ influences the development of PAO1 BLS. Bacteria were inoculated in ASM+ containing 5 mg/ml (1X), 2.5 mg/ml (0.5X), or 10 mg/ml (2X) mucin and incubated for 3 d under 20% EO2/static conditions. The structures were analyzed by CLSM and 3-D images were constructed. Architecture of PAO1 BLS formed in the presence of 1X (A), 0.5X (B), or 2X (C) mucin. Boxes, 800.00 px W x 600.00 px H; bars, 100 px. (D) After 3 d, the gelatinous mass was removed from each well and vortexed to suspend the bacteria. The bacterial suspension was serially diluted and aliquots from each dilution were spotted on LB agar to determine the CFU/ml. Values represent the means of at least three independent experiments ± SEM.