Upconverting phosphors as an amplifier of a colorimetric signal in dry chemistry test strips for enzymatic measurement of glucose.

We describe the use of upconverting phosphors (UCPs) as an amplifier of the colorimetric signal in enzymatic dry chemistry test strips for the determination of glucose. The indicator compound used in the strips attenuated both the emission and excitation wavelengths of the upconverting phosphors, thus taking advantage of the nonlinear relationship between the excitation and emission of these particles. By monitoring the emission of the UCPs under 978 nm excitation, we were able to obtain significantly higher signal response (a steeper calibration curve) from the colorimetric assay compared to a reflectance measurement. The limit of detection of the developed method was 2.1-fold lower than the reflectance-based reference method. The new method was able to determine glucose from a sample of water in the range of 5.7 µM to 22 mM, which covers the typical blood glucose range in newborns and adults. In low analyte concentrations we obtained up to 3-fold improvement in the slope of the assay calibration curve. This difference decreased with increasing analyte concentration and the effect was diminished by the highest glucose concentration. However, the analytical sensitivity (the ratio of slope and standard deviation) was practically identical between the two methods because of the higher deviation in the UCP emission signal.

Ligand enabling visible wavelength excitation of europium(III) for fluoroimmunoassays in aqueous micellar solutions.

Fluorescent reporters based on lanthanide ions, such as europium chelates, enable highly sensitive detection in immunoassays and other ligand binding assays. Unfortunately they normally require UV-excitation produced by a xenon flash or nitrogen laser light source. In order to use modern solid state excitation sources such as light emitting diodes (LEDs), these reporters need to be excited at wavelengths longer than 365 nm, where high-powered ultraviolet LEDs are available. A novel ligand, 9-ethyl-3,6-bis(5',5',5',4',4'-pentafluoro-1',3'-dioxopentyl)carbazole (bdc), was synthesized to efficiently excite europium(III) at wavelengths up to 450 nm in micellar solutions, and its performance was compared to a commercially available DELFIA enhancement solution. The detection limit of Eu(III) with the bdc-ligand using 365 nm excitation was determined to be 63 fM, which is 3 times lower than with the DELFIA solution. The bdc-ligand enabled sensitive detection of europium(III) ions in solution using 365 nm excitation and displayed similar sensitivity and functionality as commercially available DELFIA enhancement solution. Therefore, this novel enhancement solution might be a feasible alternative in producing time-resolved fluorescence under LED-excitation.

Oligonucleotide array-in-well platform for detection and genotyping human adenoviruses by utilizing upconverting phosphor label technology.

We have developed a robust array-in-well test platform based on an oligonucleotide array, combining advantages of simple instrumentation and new upconverting phosphor reporter technology. Upconverting inorganic lanthanide phosphors have a unique property of photoluminescence emission at visible wavelengths under near-infrared excitation. No autofluorescence is produced from the sample or support material, enabling a highly sensitive assay. In this study, the assay is performed in standard 96-well microtiter plates, making the technique easily adaptable to high-throughput analysis. The oligonucleotide array-in-well assay is employed to detect a selection of ten common adenovirus genotypes causing human infections. The study provides a demonstration of the advantages and potential of the upconverting phosphor-based reporter technology in multianalyte assays and anti-Stokes photoluminescence detection with an anti-Stokes photoluminescence imaging device.