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Significance: For the first time to our knowledge, a millifluidic device hasbeen designed to observe the effects of blue-light irradiation in Pseudomonasaeruginosa PA01 bacterial biofilm. Approach: A bacterial biofilm is formedunder controlled flow of nutrients in microfabricated channels settled on amicroscope stage. The setup is devised to apply defined irradiation andevaluate the photo-killing efficiency in situ using video-microscopy andfluorescent probes. Results: We investigated bacterial survival afterdelivering a precise, controlled and spatially defined blue light dose upongrowing biofilm. We demonstrate in this report that the combined use ofconstitutive GFP and propidium iodide in a millifluidic setup enables toevidence light-induced mortality at the single cell and community level.Conclusions: Real-time monitoring of a bacterial biofilm growing in amillifluidic device after irradiation opens an avenue for a betterunderstanding of the biofilm community response to blue-light, permittingresearchers to approach photokilling efficiency of biofilm as a dynamicprocess.

Design of a millifluidic device for the evaluation of blue-light disinfection on Pseudomonas aeruginosa PA01 biofilm