No abstract

Surfactants, mimics of contamination, play an important role in nanobubble nucleation, stability, and growth at the electrode surface. Herein, we utilize single-molecule fluorescence microscopy as a sensitive imaging tool to monitor nanobubble dynamics in the presence of a surfactant. Our results show that the presence of anionic and nonionic surfactants increase the rate of nanobubble nucleation at all potentials in a voltage scan. The fluorescence and electrochemical responses indicate the successful lowering of the critical gas concentration needed for nanobubble nucleation across all voltages. Furthermore, we demonstrate that the accumulation of surfactants at the gas–liquid interface changes the interaction of fluorophores with the nanobubble surface. Specifically, differences in fluorophore intensity and residence lifetime at the nanobubble surface suggest that the labeling of nanobubbles is affected by the nature of the nanobubble (size, shape, etc.) and the structure of the gas–liquid interface (surfactant charge, hydrophobicity, etc.).

This work provides insight into the mechanism of autoclave killing of spores of an organism used in BIs, and that a spore enzyme in a BI is more stable to autoclaving than spore viability.

Berberine, an alkaloid originally extracted from the plant Coptis chinensis and other herb plants, has been used as a pharmacological substance for many years. The therapeutic effect of berberine has been attributed to its interaction with nucleic acids and blocking cell division. However, levels of berberine entering individual microbial cells minimal for growth inhibition and its effects on bacterial spores have not been determined. In this work the kinetics and levels of berberine accumulation by individual dormant and germinated spores were measured by laser tweezers Raman spectroscopy and differential interference and fluorescence microscopy, and effects of berberine on spore germination and outgrowth and spore and growing cell viability were determined. The major conclusions from this work are that: (1) colony formation from B. subtilis spores was blocked ~ 99% by 25 μg/mL berberine plus 20 μg/mL INF55 (a multidrug resistance pump inhibitor); (2) 200 μg/mL berberine had no effect on B. subtilis spore germination with L-valine, but spore outgrowth was completely blocked; (3) berberine levels accumulated in single spores germinating with ≥ 25 μg/mL berberine were > 10 mg/mL; (4) fluorescence microscopy showed that germinated spores accumulated high-levels of berberine primarily in the spore core, while dormant spores accumulated very low berberine levels primarily in spore coats; and (5) during germination, uptake of berberine began at the time of commitment (T1) and reached a maximum after the completion of CaDPA release (Trelease) and spore cortex lysis (Tlysis).

In schematics and cartoons, the gas–liquid interface is often drawn as solid lines that aid in distinguishing the separation of the two phases. However, on the molecular level, the structure, shape, and size of the gas–liquid interface remain elusive. Furthermore, the interactions of molecules at gas–liquid interfaces must be considered in various contexts, including atmospheric chemical reactions, wettability of surfaces, and numerous other relevant phenomena. Hence, understanding the structure and interactions of molecules at the gas–liquid interface is critical for further improving technologies that operate between the two phases. Electrochemically generated surface nanobubbles provide a stable, reproducible, and high-throughput platform for the generation of a nanoscale gas–liquid boundary. We use total internal reflection fluorescence microscopy to image single-fluorophore labeling of surface nanobubbles in the presence of a surfactant. The accumulation of a surfactant on the nanobubble surface changes the interfacial properties of the gas–liquid interface. The single-molecule approach reveals that the fluorophore adsorption and residence lifetime at the interface is greatly impacted by the charge of the surfactant layer at the bubble surface. We demonstrate that the fluorescence readout is either short- or long-lived depending on the repulsive or attractive environment, respectively, between fluorophores and surfactants. Additionally, we investigated the effect of surfactant chain length and salt type and concentration on the fluorophore lifetime at the nanobubble surface.