Figure 3: 3D fluorescence microscopic visualisation of PAO1 biofilms grown in different conditions (3a) and corresponding biovolumes (3b). PAO1 biofilms were performed in 3 well chambered cover glasses under 2.5 ml h-1 flow. LB (Control), LB with 0.1% or 1% ECO or LB with 0.1% or 1% EOB emulsified with ECO in the same amount, in relation to the EOB concentrations, were used as growth media. No biofilm was observed at 1% EOB concentration (image not shown). Total biofilm cells were detected after SYTO9 (5 μM) staining and appear in green, as altered cells stained with Propidium Iodid (PI, 0.3 μM) appear in red. The overlay represents the superposition of SYTO9 and PI stained cells images. Scale represents 67.3 μm. Total biovolume and maximal thickness (Tm) were obtained from SYTO9 stained cell images treated with COMSTAT. Mortality rates (M) were obtained by calculating the ratio: total biovolume (SYTO9) / altered biovolume (PI) x 100. Error bars represent standard deviation from the mean (n=9) and *P<0.05, ***P<0.001.