Ion-detecting microspheres
View Patent ↗This invention provides methods of using ion-detecting microspheres containing an ionphore and a chromoionphore in clinical laboratory instrumentation such as flow cytometry for sample analysis. In one embodiment, the microspheres are contacted with a flowing stream of a sample under conditions that allow the ion-selective ionophores to complex with the ions in the sample, and to cause deprotonation of the chromoionophore. The complexes are then exposed to an excitation wavelength light source suitable for exciting the deprotonated chromoionophore to emit a fluorescence signal pattern. Detection of the fluorescence signal pattern emitted by the deprotonated chromoionophore in microspheres containing the complexes allows for determination of the presence of the target ions in the sample. In one embodiment, lead ion-detecting microspheres are provided that can detect nanomolar levels of lead ions with response times on the order of minutes.
1. Ion-detecting polymeric microspheres for detecting metal ions in a sample, said microspheres comprising a lipophilic ionophore selective for said metal ions, a chromoionophore selective for hydrogen ions, a copolymer having a glass transitional temperature (T g ) of about or below 0° C., and a flourescent dye, wherein said chromoionophore is a chromoionophore that becomes deprotonated when said metal ions are present in said sample, and wherein said deprotonated form of said chromoionophore is absorbent at the frequency of the fluorescence emission of said dye, wherein said copolymer comprises polymerized units of methacrylate monomers, wherein said methacrylate monomers comprise a monomer having an R 1 pendant alkyl group and a monomer having an R 2 pendant alkyl group, wherein R 1 is any of C 1-3 alkyl groups and R 2 is any of C 4-12 alkyl groups.
2. The polymeric microspheres of claim 1 , wherein said metal ions are lead ions and said ionophore is N,N,N′,N′-tetradodecyl-3,6-dioxaoctane-1-thio-8-oxodiamide (ETH 5493).
3. The ion-detecting sensor of claim 2 , wherein said chromoionophore is 11-[(butylpentyl)oxy]-11-oxoundecyl-4-[9-(dimethylamino)5H-benzi[a]phenoxazin-5-yl-indene]aminobenzoate (ETH 5418) and said dye is 1,1″-dioctadecyl-3,3,3′,3′-tetramethylindorcarbocyanine perchlorate (DiIC 18 ).
4. The polymeric microspheres of claim 1 , wherein said polymer further comprises an ion-exchanger.
5. The polymeric microspheres of claim 1 , wherein said particles are obtained by an automated casting process.