Figure 1
NBD-UBIdend displays fluorescence amplification in hydrophobic environments and labels a diverse panel of clinically relevant pathogens in a concentration dependent manner. (A) The structure of NBD-UBI (monomer). (B) The structure of NBD-UBIdend (multivalent construct). (C) NBD-UBIdend demonstrates fluorescence amplification in increasing concentrations of t-butanol mimicking a hydrophobic environment (representative plot, n = 3). (D) Fluorescence quantification of P. aeruginosa imaged on a benchtop confocal microscope in the continual presence of increasing concentrations of NBD-UBIdend. Images show representative images at denoted concentrations of NBD-UBIdend (scale bar represents 5 μm). Each point on the graph represents the mean (+/− SEM) of three independent experiments where at least three fields of view were quantified with a single site non-linear fit of the data. (E) Representative confocal images of a panel of clinically relevant pathogens imaged in the continued presence of NBD-UBIdend (5 μM), scale bar represents 5 μm. P. aeruginosa data includes both the laboratory strain (PA01) and a clinical isolate from a patient with VAP (J3284). (F) Quantification of fluorescent intensity of bacterial imaging on a benchtop confocal microscope. Data shows the mean (+/− SEM) of three independent experiments where at least three independent fields of view were assessed.
