Selective Optical Imaging for Detection of Bacterial Biofilms in Tissues.

SIGNIFICANCE: The development of an imaging technique to accurately identify biofilm regions on tissues and in wounds is crucial for the implementation of precise surface-based treatments, leading to better patient outcomes and reduced chances of infection. AIM: The goal of this study was to develop an imaging technique that relies on selective trypan blue (TB) staining of dead cells, necrotic tissues, and bacterial biofilms, to identify biofilm regions on tissues and wounds. APPROACH: The study explored combinations of ambient multi-colored LED lights to obtain maximum differentiation between stained biofilm regions and the underlying chicken tissue or glass substrate during image acquisition. The TB imaging results were then visually and statistically compared to fluorescence images using a shape similarity measure. RESULTS: The comparisons between the proposed TB staining method and the fluorescence standard used to detect biofilms on tissues and glass substrates showed up to 97 percent similarity, suggesting that the TB staining method is a promising technique for identifying biofilm regions. CONCLUSIONS: The TB staining method demonstrates significant potential as an effective imaging technique for the identification of fluorescing and non-fluorescing biofilms on tissues and in wounds. This approach could lead to improved precision in surface-based treatments and better patient outcomes.

Fabrication and Performance of a Multi-Discharge Cold Atmospheric Pressure Plasma Array.

Cold atmospheric pressure plasma (CAP) has been shown to kill bacteria and remove biofilms. Here we report the development of a unique CAP array device consisting of a parallel stack of eight linear-discharge plasma elements that create a ~ 5 cm2 (2.4 cm × 2 cm) treatment area. The CAP device is fabricated from Low Temperature Co-fired Ceramic (LTCC) layers to create 24 mm long linear-discharge channels (500 µm gap) with embedded opposing silver metal electrodes. A 20 kHz AC voltage (0.5-5 kV) applied to the electrodes generates an Ar/O2 plasma between the plates, with the gas flow directing the reactive species toward the biological sample (biofilms, etc.) to affect the antimicrobial treatment. External ballast resistors were used to study discharge uniformity in the stacked array elements and internal thick film ballast resistors (≈150 kΩ) were developed to create a fully integrated device. Typical element discharge currents were 1-2.5 mA with the total array current tested at 20 mA to provide optimal device uniformity. The plasma discharge was further shown to produce reactive hydrogen peroxide and exert antimicrobial effects on Pseudomonas biofilms and Salmonella contaminated eggshell samples, with >99% of the bacterial cells killed with less than 60 seconds of plasma exposure.