Luminescent CeO2:Eu3+ nanocrystals for robust in situ H2O2 real-time detection in bacterial cell cultures

Hydrogen peroxide (H2O2) quantification in biomedicine is valuable as inflammation biomarker but also in assays employing enzymes that generate or consume H2O2 linked to a specific biomarker. Optical H2O2 detection is typically performed through peroxidase-coupled reactions utilizing organic dyes that suffer, however, from poor stability/reproducibility and also cannot be employed in situ in dynamic complex cell cultures to monitor H2O2 levels in real-time. Here, we utilize enzyme-mimetic CeO2 nanocrystals that are sensitive to H2O2 and study the effect of H2O2 presence on their electronic and luminescent properties. We produce and dope with Eu3+ these particles in a single-step by flame synthesis and directly deposit them on Si and glass substrates to fabricate nanoparticle layers to monitor in real-time and in situ the H2O2 concentrations generated by Streptococcus pneumoniae clinical isolates. Furthermore, the small CeO2:Eu3+ nanocrystals are combined in a single-step with larger, non-responsive Y2O3:Tb3+ nanoparticles during their double-nozzle flame synthesis to engineer hybrid luminescent nanoaggregates as ratiometric robust biosensors. We demonstrate the functionality of these biosensors by monitoring their response in the presence of a broad range of H2O2 concentrations in vitro from S. pneumoniae, highlighting their potential for facile real-time H2O2 detection in vitro in cell cultures.