Microscale plasticized PVC particles doped with hydrophobic ionophores are prepared by solvent evaporation of aqueous suspensions of sensing cocktails (poly(vinyl chloride), plasticizer, active sensing components, and tetrahydrofuran) and tested as particulate microoptical sensors. The particles contain either only the chromoionophore ETH 2458 as active reagent or the potassium ionophore BME-44, chromoionophore ETH 5294, and lipophilic anionic sites NaTFPB. The former system functions according to an anion-hydrogen ion coextraction mechanism and shows a Hofmeister anion selectivity pattern, while the latter sensor containing additional ionophores represents the more complex, truly selective optical sensors based on ion-exchange equilibria. Single microspheres are simultaneously characterized spatially and spectrally by fluorescence microscopy, coupled to a spectrometer equipped with a CCD detector. The results indicate that these microspheres respond in complete analogy to traditional thin-film-based optodes previously reported in the literature. The introduction of small, spherical ionophore-based sensing particles that operate on the basis of bulk extraction principles holds the promise of significantly expanding the available chemical palette of microsphere-based analytical assays.

The use of flow cytometry as a rapid, high-throughput diagnostic tool for the analysis of ions is described. Monodisperse, uniform microspheres, which obey bulk optode theory and are governed by bulk extraction processes rather than surface phenomena, were prepared under mild, nonreactive conditions using a sonic stream particle casting apparatus. As an initial example demonstrating the utility of this approach, microspheres that contained a H+-selective fluorescent chromoionophore (ETH 5294), a cation-exchanger (NaTFPB), and either a highly sodium-selective (sodium ionophore X) or a potassium-selective ionophore (BME-44) were prepared. Separate solution analysis of sodium- and potassium-selective microspheres resulted in the generation of functional response curves using peak channel fluorescence intensities. The selectivity observed for both types of particles is sufficient for the clinical determination of Na+ and K+. Furthermore, sodium- and potassium-selective microspheres were analyzed in parallel using sodium sample solutions, resulting in the successful determination of sodium ion concentrations and providing important information about the selectivity of the potassium-selective sensors over sodium. This work demonstrates the potential applicability of flow cytometry as a means for developing multiplexed, rapid, high-throughput analyses for clinically relevant ions.

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