BioEISense: A Microfluidic Platform for Real‐Time Monitoring of Staphylococcus aureus Biofilm Formation and the Efficacy of Antibiofilm Agents

ABSTRACT Antimicrobial resistance (AMR), largely triggered by bacterial biofilms, is a major threat to global health. Bacteria within biofilms exhibit up to 1000‐fold higher AMR than in their planktonic form. Standard biofilm analysis methods rely on static well‐plate assays and destructive endpoints. Therefore, new methods enabling dynamic culture conditions and real‐time measurements are urgently needed to better understand biofilm evolution and eradication. This study presents BioEISense, a microfluidic biofilm sensing platform, which incorporates a flow‐through biofilm culture channel made from off‐stoichiometric thiol‐ene and polydimethylsiloxane polymers (bottom and cover, respectively). In addition, the BioEISense has integrated gold sensors for noninvasive monitoring of biofilm formation by electrochemical impedance spectroscopy (EIS) in real‐time. The performance of BioEISense is first optimized in terms of electrode design, bacterial concentration, and culture media, using EIS. The EIS results are additionally validated by measuring biofilm thickness using confocal microscopy. Finally, the feasibility of BioEISense for screening of the efficacy of antibiofilm agents is evaluated using both biofilm prevention and eradication assays. Together, our findings highlight the great competence of microfluidic biofilm assays for real‐time, noninvasive monitoring of biofilm growth and degradation, with a view to discovering new antibiofilm agents to combat AMR.