A1r live cell inverted confocal microscope

Established biofilms of strains JSG1149 and JSG1150 were incubated with or without ciprofloxacin for 8 hours prior to imaging. Biofilms were washed two times in PBS and fixed in 2% paraformaldehyde (PFA, Affimetrix, Cleveland, OH) for 20 minutes at room temperature at each time point and saved for later imaging. The amount of biofilm and the structure of the biofilm can be inferred by the amount of GFP signal detected⁴⁸ . Biomass and average thickness were assessed by automated capturing of 10 random Z-stacks per well in 5 wells per treatment using a Nikon A1R Live Cell Inverted Confocal microscope. The Z-stacks were then analyzed using the software package COMSTAT2⁴⁹ .

Established biofilms of GFP-expressing strains JSG4093 and JSG4244 were incubated with or without 0.2% L-arabinose for 24 hr prior to imaging. Biofilms were washed two times in PBS and fixed in 2% paraformaldehyde (PFA, Affimetrix) for 1 hour at room temperature and saved for later imaging. The amount of biofilm and the structure of the biofilm can be inferred by the amount of GFP signal detected (Franklin et al., 2015 (link)). Biomass and average thickness were assessed by automated capturing of 5 random Z-stacks per well in 4 wells per treatment using a Nikon A1R Live Cell Inverted Confocal microscope. The Z-stacks were then analyzed using the software package COMSTAT2.