Biofilm development of an opportunistic model bacterium analysed at high spatiotemporal resolution in the framework of a precise flow cell.

Life of bacteria is governed by the physical dimensions of life in microscales, which is dominated by fast diffusion and flow at low Reynolds numbers. Microbial biofilms are structurally and functionally heterogeneous and their development is suggested to be interactively related to their microenvironments. In this study, we were guided by the challenging requirements of precise tools and engineered procedures to achieve reproducible experiments at high spatial and temporal resolutions. Here, we developed a robust precise engineering approach allowing for the quantification of real-time, high-content imaging of biofilm behaviour under well-controlled flow conditions. Through the merging of engineering and microbial ecology, we present a rigorous methodology to quantify biofilm development at resolutions of single micrometre and single minute, using a newly developed flow cell. We designed and fabricated a high-precision flow cell to create defined and reproducible flow conditions. We applied high-content confocal laser scanning microscopy and developed image quantification using a model biofilm of a defined opportunistic strain, Pseudomonas putida OUS82. We observed complex patterns in the early events of biofilm formation, which were followed by total dispersal. These patterns were closely related to the flow conditions. These biofilm behavioural phenomena were found to be highly reproducible, despite the heterogeneous nature of biofilm.

Generation of photocurrents by bis-aniline-cross-linked Pt nanoparticle/photosystem I composites on electrodes.

Pt nanocrystals are implanted into photosystem I (PSI) by a photochemical reaction. The PSI with the associated Pt nanoclusters was modified with thioaniline and electropolymerized with thioaniline-functionalized Pt nanoparticles (NPs) to yield a bis-aniline-cross-linked PSI/Pt NPs composite. The alignment of the PSI with respect to the Pt NPs leads to effective charge separation and to generation of a photocurrent, ϕ (λ = 420 nm) = 2.6%, IPCE ∼ 0.35%. The bis-aniline units cross-linking the PSI/Pt NPs composite exhibit quasireversible redox features (E(0)' = 0.05 V vs Ag/AgCl, at pH = 7.4). Biasing the electrode potential, E > 0.1 V vs SCE, results in the formation of the oxidized quinoid bis-aniline state that acts as an electron acceptor. At this applied potential, the bridges mediate the electron transfer to the electrode, resulting in a ∼10-fold higher photocurrent, as compared to the system that includes the reduced bis-aniline bridging units. Furthermore, the ferredoxin (Fd) electron relay was modified with thioaniline units and incorporated into the PSI/Pt NP composite during the electropolymerization process. The Fd bound to the matrix mediates the electron transfer toward the electrode and facilitates charge separation that results in enhanced generation of the photocurrent, ϕ (λ = 420 nm) = 3.8%, IPCE ∼ 0.5%.