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Abstract Biofilms, notorious for their recalcitrance and dynamic behavior, pose a persistent threat to public health. However, existing diagnostic tools fall short in providing in situ, spatiotemporal biochemical insights into dynamic biofilm behavior. To address this, we have developed zwitterionic nanoplasmonic bio-meshes that combine the antifouling attributes of zwitterionic L-cysteine, the biocompatibility of polymeric meshes, and the ultrasensitive, uniform, and stable surface-enhanced Raman spectroscopy (SERS) response of plasmonic nanocavity arrays. This platform delivers improved SERS performance in human serum compared to controls without L-cysteine functionalization, achieving a clinically-relevant limit of detection of 5.6 nM for pyocyanin in undiluted human serum. Moreover, the platform enables real-time, in situ spatiotemporal SERS monitoring ofP. aeruginosabiofilms over 48 h in culture media-agar backgrounds, revealing distinct pyocyanin secretions dynamics in wild-type and hyperbiofilm mutant strains. We envision that this capability to non-invasively monitor biofilm metabolite secretion dynamics can empower next-generation biofilm diagnostics and anti-biofilm therapies. 

This study presents antifouling nanoplasmonic bio-meshes forin situspatiotemporal SERS monitoring ofP. aeruginosabiofilms. The bio-meshes combine zwitterionic L-cysteine antifouling properties with biocompatible polymeric meshes and sensitive SERS nanocavity arrays. This platform enables ultrasensitive pyocyanin detection in undiluted serum and reveals pyocyanin dynamics inP. aeruginosabiofilms, highlighting the link between metabolite secretion and cyclic di-GMP signaling. 

Nanolaminate nano-optoelectrodes can generate plasmon-enhanced electronic and vibrational Raman scattering signals to probe transition state dynamics during electrochemical redox reactions based on voltage-dependent spectroscopic signatures of interfacial electronic and molecular states at plasmonic hotspots. 

This study explores dynamic voltage-modulated nanoplasmonic metal luminescence at the electrode-electrolyte interface using nanolaminate nano-optoelectrode arrays. We demonstrate a negative voltage modulation slope in nanoplasmonic metal luminescence during electrochemical surface-enhanced Raman spectroscopy measurements. 

We introduce scalable two-tier protruding micro-/nano-optoelectrode arrays, integrating biophotonic and bioelectronic functionalities with high nanoplasmonic Raman enhancement and reduced interfacial impedance, crucial forin situbioelectrical and biochemical measurements in multicellular systems. 

This study introduces biomimetic transparent nanoplasmonic microporous mesh (BTNMM) devices fabricated via reverse nanoimprint lithography. These devices offer spatiotemporal multimodal SERS measurements for bio-interfaced applications, enabling targeted pH sensing and molecular profiling of microbial biofilms.