Analyzing Pseudomonas Biofilm Disruption

P. aeruginosa biofilms were grown in LB medium supplemented with 0.2% glucose at 20 °C in flow cell chambers as was previous described^{56 (link),57 (link)}. After 96 h the mature biofilm was treated with different alginate lyases for 12 h. For analysis, the biofilm was stained using the LIVE/DEAD BacLight Bacterial Viability kit (Thermo Scientific) and visualized with a Zeiss LSM 800 confocal laser scanning microscope (CSLM) using the 20×/0.8 objective with excitation wavelengths of 488 and 560 nm. Microscope images were processed with the ImageJ analysis software and COMSTAT 2, a specific biofilm analysis software was used to quantify the biofilm biomass, thickness and roughness^{58 (link)}.