# Trypan Blue Image-Guided Removal of Surface-Based Bacterial Biofilms from Chicken Tissue Using Cold Atmospheric Pressure Plasma

## TL;DR

This study shows that cold plasma can effectively remove bacterial biofilms from chicken tissue with high efficiency and minimal heat.

## Contribution

A novel image-guided cold plasma method for biofilm removal is introduced with precise operational parameters and performance metrics.

## Key findings

- CAP treatment achieved >99% biofilm reduction with an etch rate of 2.2–5.8 μm/s.
- Biofilm thickness on chicken tissue ranged from 20 to 180 μm, similar to mammalian tissues.
- Non-thermal disruption was indicated as the plasma electrode reached 94.7 °C while the biofilm area reached 36.3 °C.

## Abstract

The study evaluates the efficacy of an image-guided CAP treatment method with a plasma device capable of rapid biofilm removal from chicken tissue. The plasma treatment operating configuration includes a gas mixture of Argon and H2O at a flowrate of 1.5 lpm. An X-Y stage was used to move the chicken sample below the stationary plasma scalpel at a speed of 0.1 mm/s. The discharge voltage and current were maintained between 3.2 and 3.7 kV (AC 20 kHz), and at 3 mA, respectively. The electrode gap and sample distance were set to 0.6 mm and 4 mm. This configuration facilitated effective biofilm removal, as confirmed by CFU analysis and 3D microscopic analysis showing a >99% reduction in biofilm post treatment with an etch rate of 2.2–5.8 μm/s and an impact width of up to 300 μm. The plasma scalpel electrode temperature reached 94.7 °C, while the targeted biofilm area was heated to 36.3 °C, suggesting non-thermal biofilm disruption. Three-dimensional microscopic analysis revealed biofilm thickness on chicken tissues ranging from 20 to 180 μm, comparable to biofilm loads on mammalian tissues. In conclusion, the study highlights the potential of CAP devices as a promising solution for biofilm debridement.