Decoupling Surgical Bioaerosol Burden and Biological Activity with Point-of-Care Microfluidic-Plasmonic Biosensing

Point-of-care bioaerosol analysis remains a critical gap in perioperative exposure assessment, as existing air-monitoring approaches primarily quantify particle burden while providing limited information on aerosol-associated biomolecular composition. Here, we present Surgical Personalized On-site Tracker (SPOT), a portable microfluidic sampling-to-biosensing system with localized surface plasmon resonance (LSPR) biosensing for dual-parameter assessments of proteinaceous aerosol burden and accessible adenosine triphosphate (ATP). The SPOT platform enables on-site detection of total protein as a surrogate for aerosolized biological loads, whereas the ATP channel quantifies accessible ATP after aerosol collection and sample processing without intentional cell lysis. In a clinical study conducted during thoracic surgeries, SPOT detected markedly elevated bioaerosols compared with nonsurgical conditions, with total proteinaceous bioaerosol burden levels reaching 39.93 pg/m³ and accessible ATP concentrations up to 78.97 pmol/m³. Importantly, minimally invasive procedures exhibited a significantly higher ATP-to-protein ratio than open surgeries, i.e., 3.15 vs 2.07 pmol/pg, revealing disproportionally elevated accessible ATP within surgical plumes despite lower total mass. These findings established ATP-to-protein profiling as a novel metric for functional bioaerosol surveillance and demonstrated the potential of SPOT for perioperative air-quality monitoring and biosafety evaluation.