Fluorescent pH detector system and related methods
First Claim
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1. A method for measuring the pH of a sample, comprising:
- (a) irradiating a fluorescent species immobilized on a substrate with excitation light emanating from a probe physically isolated from the fluorescent species immobilized on the substrate, wherein the fluorescent species immobilized on the substrate is in liquid communication with a sample, wherein the excitation light has a wavelength sufficient to effect fluorescent emission from the fluorescent species, wherein the fluorescent species exhibits a first emission intensity at a first emission wavelength and a second emission intensity at a second emission wavelength, the ratio of the first and second emission intensities being dependent on pH; and
(b) measuring the first and second emission intensities to determined the pH of the sample.
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Abstract
Fluorescent pH detector and methods for measuring pH using the fluorescent pH detector.
120 Citations
26 Claims
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1. A method for measuring the pH of a sample, comprising:
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(a) irradiating a fluorescent species immobilized on a substrate with excitation light emanating from a probe physically isolated from the fluorescent species immobilized on the substrate, wherein the fluorescent species immobilized on the substrate is in liquid communication with a sample, wherein the excitation light has a wavelength sufficient to effect fluorescent emission from the fluorescent species, wherein the fluorescent species exhibits a first emission intensity at a first emission wavelength and a second emission intensity at a second emission wavelength, the ratio of the first and second emission intensities being dependent on pH; and
(b) measuring the first and second emission intensities to determined the pH of the sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A system for measuring pH, comprising:
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(a) a light source for exciting a fluorescent species, wherein the fluorescent species has a first emission intensity at a first emission wavelength and a second emission intensity at a second emission wavelength;
(b) a first emission detector for measuring the first emission intensity;
(c) a second emission detector for measuring the second emission intensity;
(d) an excitation lightguide for transmitting excitation light from the light source to the fluorescent species, wherein the lightguide comprises a first terminus proximate to the light source and a second terminus distal to the light source;
(e) a first emission lightguide for transmitting emission from the fluorescent species to the first emission detector, wherein the lightguide comprises a first terminus proximate to the detector and a second terminus distal to the detector;
(f) a second emission lightguide for transmitting emission from the fluorescent species to the second emission detector, wherein the lightguide comprises a first terminus proximate to the detector and a second terminus distal to the detector;
(g) a probe housing the distal termini of the excitation lightguide, first emission lightguide, and second emission light guide; and
(h) an assembly for receiving the probe, the assembly comprising;
(i) a housing for receiving the probe, wherein the housing is adapted for receiving the probe at a first end and terminating with a window at the second end, the window being transparent to the excitation and the emission light, (ii) a tip member reversibly connectable to the housing'"'"'s second end, wherein the tip member is adapted to receive liquid from a sample to be measured, and (iii) a fluorescent species immobilized on a substrate intermediate the tip member and the window, wherein the fluorescent species immobilized on the substrate is in liquid communication with the sample during the measurement, and wherein the window physically isolates the probe member from the fluorescent species immobilized on the substrate. - View Dependent Claims (13, 14, 15, 16, 17, 18)
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19. An assembly, comprising
(a) a housing having a first open end and a second closed end, wherein the closed end comprises a window transparent to visible light; -
(b) a tip member reversibly connectable to the housing closed end, wherein the tip member is adapted to expose the housing window; and
(c) a fluorescent species immobilized on a substrate intermediate the housing window and tip member. - View Dependent Claims (20)
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- 21. A conjugate, comprising an environment-sensitive fluorophore covalently coupled to an albumin.
- 23. A membrane-immobilized fluorescent species, comprising a conjugate of a naphthofluorescein compound and an albumin adhered to a membrane.
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25. A method for measuring the carbon dioxide level in a liquid sample, comprising:
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(a) irradiating a fluorescent species immobilized on a substrate with excitation light emanating from a probe physically isolated from the fluorescent species immobilized on the substrate, wherein the fluorescent species immobilized on the substrate is in liquid communication with a solution having pH responsiveness to carbon dioxide present in a liquid sample, wherein the solution having pH responsiveness to carbon dioxide is in communication with the liquid sample through a selectively permeable membrane, wherein the excitation light has a wavelength sufficient to effect fluorescent emission from the fluorescent species, wherein the fluorescent species exhibits a first emission intensity at a first emission wavelength and a second emission intensity at a second emission wavelength, the ratio of the first and second emission intensities being dependent on pH;
(b) measuring the first and second emission intensities to determined the pH of the solution having pH responsiveness; and
(c) correlating the pH of the solution having pH responsiveness to the carbon dioxide level in the sample.
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26. A system for measuring carbon dioxide level in a liquid sample, comprising:
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(a) a light source for exciting a fluorescent species, wherein the fluorescent species has a first emission intensity at a first emission wavelength and a second emission intensity at a second emission wavelength;
(b) a first emission detector for measuring the first emission intensity;
(c) a second emission detector for measuring the second emission intensity;
(d) an excitation lightguide for transmitting excitation light from the light source to the fluorescent species, wherein the lightguide comprises a first terminus proximate to the light source and a second terminus distal to the light source;
(e) a first emission lightguide for transmitting emission from the fluorescent species to the first emission detector, wherein the lightguide comprises a first terminus proximate to the detector and a second terminus distal to the detector;
(f) a second emission lightguide for transmitting emission from the fluorescent species to the second emission detector, wherein the lightguide comprises a first terminus proximate to the detector and a second terminus distal to the detector;
(g) a probe housing the distal termini of the excitation lightguide, first emission lightguide, and second emission light guide; and
(h) an assembly for receiving the probe, the assembly comprising;
(i) a housing for receiving the probe, wherein the housing is adapted for receiving the probe at a first end and terminating with a window at the second end, the window being transparent to the excitation and the emission light, (ii) a tip member reversibly connectable to the housing'"'"'s second end, wherein the tip member comprises a chamber for receiving a solution having pH responsiveness to carbon dioxide present in a liquid sample, wherein the solution having pH responsiveness to carbon dioxide is in liquid communication with the liquid sample through a selectively permeable membrane, and (iii) a fluorescent species immobilized on a substrate intermediate the tip member and the window, wherein the fluorescent species immobilized on the substrate is in liquid communication with the solution having pH responsiveness to carbon dioxide during the measurement, and wherein the window physically isolates the probe member from the fluorescent species immobilized on the substrate.
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Specification