The objective of this study was to benchmark the different signal enhancements found in a recently exploited fluorescence ratiometric optode design used for nano-molar imaging of ammonium and ammonia. The sensing scheme of these optodes are based on a mediated transfer of the analyte together with a fluorescent dye in a two-phase system consisting of a gold nanoparticle (GNP) doped ether (organic phase), emulsified in a hydrogel (hydrous phase). The coextraction of the ion dye pair causes changes in fluorescence in relation to the analyte concentration. Performances of optodes with and without GNPs using the gradually improved instrumentation and signal processing were evaluated and normalized to be comparable. Signal to noise was enhanced due to signal processing based on ensemble averaging (1.7× - 3.2×), CCD sensitivity (2×), and plasmon assisted fluorescence (10× - 100×), which altogether with the ratiometric treatment of the fluorescence contributed to the great sensitivity for ammonium and ammonia. The study shows that GNP doped sensors are relatively more sensitive to matrix effects but if they are isolated by a protective layer they will dramatically increase in sensitivity. Proper isolation of the active chemical components from the matrix will make the sensor design one of the most powerful and versatile concepts for chemical imaging and single point detection in complex environments as the optodes likely can be constructed for most ions that have selective ionophores.
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