Real-time monitoring of the dynamics and interactions of bacteria and the early-stage formation of biofilms

Bacterial biofilms are complex colonies of bacteria adhered to a static surface and/or one to another. Bacterial biofilms exhibit high resistance to antimicrobial agents and can cause life-threatening nosocomial infections. Despite the effort of the scientific community investigating the formation and growth of bacterial biofilms, the preliminary interaction of bacteria with a surface and the subsequent early-stage formation of biofilms is still unclear. In this study, we present real-time, label-free monitoring of the interaction of Escherichia coli and Pseudomonas aeruginosa bacteria with untreated glass control surfaces and surfaces treated with benzalkonium chloride, a chemical compound known for its antimicrobial properties. The proof of principle investigation has been performed in a standard inverted optical microscope exploiting the optical phenomenon of caustics as a tool for monitoring bacterial diffusion and early adhesion and associated viability. The enhanced resolving power of the optical set-up allowed the monitoring and characterization of the dynamics of the bacteria, which provided evidence for the relationship between bacterial adhesion dynamics and viability, as well as the ability to form a biofilm. Viable bacteria adhered to the surface exhibited noticeable sliding or rotary dynamics while bacteria killed by surface contact remained static once adhered to the surface. This difference in dynamics allowed the early detection of biofilm formation and offers the potential to quantify the efficiency of antimicrobial surfaces and coatings.