Quantifying Biofilm Development via Flow Cells

Relative levels of biofilm development were assessed using flow cells (Kharadi & Sundin, 2019 (link)). Strains expressing GFP via vector pMP2444 (Table 1) were grown overnight and normalized at an OD₆₀₀ of 0.5. Flow cell chambers in a µ‐Slide VI (Ibidi) were inoculated with the cultures and incubated for 1 h at 24°C. Following this, a flow of 0.5 × LB medium was applied to the flow chambers using a peristaltic pump (Ismatec REGLO; Cole‐Parmer) for 5 h. Biofilms were visualized using a FluoView 1000 confocal laser‐scanning microscope (Olympus). z‐Stacked images of the flow cell channels were processed by ImageJ and compared for the green fluorescence value using the RBG assessment plug‐in (Schneider et al., 2012 (link)). Three replicates conducted in the study were statistically compared using Tukey's HSD on JMP statistical software.