Ratiometric fluorescence method to measure oxygen

Claim 1

A method for measuring oxygen in a fluid sample, comprising: (a) providing an optical sensor comprising an optical waveguide having a distal end portion for contacting the fluid sample, and a proximal end portion for communication with means for receiving a signal from the distal end portion, and wherein the distal end portion has an oxygen sensor means comprising a cross-linked oxygen-permeable membrane of a cured perfluorinated urethane polymer and, physically entrapped therein, an indicator composition of an oxygen indicator selected from the group consisting of complexes of ruthenium (II), osmium (II), iridium (III), rhodium, rhenium, and chromium (III) with 2,2'-bipyridine, 1,10-phenanthrolene, 4,7-diphenyl-1,10-phenanthrolene, 4,7-dimethyl-1,10-phenanthrolene, 4,7-disulfonated-diphenyl-1,10-phenanthrolene, 2,2'-bi-2-thiazoline, 2,2'-bithiazole, 5-bromo-1,10-phenanthrolene, and 5-chloro-1,10-phenanthrolene, and complexes of VO (II), Cu (II), platinum (II), and zinc (II) with porphyrin, etioporphyrin, tetraphenylporphyrin, mesoporphyrin IX dimethylester, protoporphyrin IX dimethylester and octaethylporphyrin, and wherein the oxygen indicator provides for an oxygen-sensitive fluorescence emission signal, and a reference dye comprising a fluorescent organic compound selected from the group consisting of fluorescein, carboxyfluorescein, hydroxypyrene trisulfonic acid, dichlorofluorescein, N,N'-bis(2,5-di-tert-butylphenyl)-3,4,9,10-perylenecarboximide, and N,N'-bis(2,6-xylidyl)-3,4,9,10-perylenecarboximide, and wherein the reference dye provides for an oxygen-insensitive fluorescence emission signal; (b) contacting the fluid sample with the distal end portion of the optical sensor; (c) exciting the indicator composition at using radiation of a first wavelength, to produce an oxygen indicator emission signal at a second wavelength and a reference dye emission signal at a third wavelength; (d) calculating the apparent quantity of oxygen present in the fluid sample from the oxygen indicator emission signal; and (e) correcting the apparent quantity of oxygen present for variations resulting from external factors, by determining the ratio of the oxygen indicator emission signal to the reference dye emission signal.

Claim 2

A method for measuring oxygen in a fluid sample, comprising: (a) providing an optical sensor comprising an optical waveguide having a distal end portion for contacting the fluid sample, and a proximal end portion for communication with means for receiving a signal from the distal end portion, and wherein the distal end portion has an oxygen sensor means comprising a cross-linked oxygen-permeable membrane of a cured perfluorinated urethane polymer and, physically entrapped therein, an indicator composition of an oxygen indicator selected from the group consisting of complexes of ruthenium (II), osmium (II), iridium (III), rhodium, rhenium, and chromium (III) with 2,2'-bipyridine, 1,10-phenanthrolene, 4,7-diphenyl-1,10-phenanthrolene, 4,7-dimethyl-1,10-phenanthrolene, 4,7-disulfonated-diphenyl-1,10-phenanthrolene, 2,2'-bi-2-thiazoline, 2,2'-bithiazole, 5-bromo-1,10-phenanthrolene, and 5-chloro-1,10-phenanthrolene, and complexes of VO (II), Cu (II), platinum (II), and zinc (II) with porphyrin, etioporphyrin, tetraphenylporphyrin, mesoporphyrin IX dimethylester, protoporphyrin IX dimethylester and octaethylporphyrin, and wherein the oxygen indicator provides for an oxygen-sensitive fluorescence emission signal, and a reference dye comprising a fluorescent organic compound selected from the group consisting of fluorescein, carboxyfluorescein, hydroxypyrene trisulfonic acid, dichlorofluorescein, N,N'-bis(2,5-di-tert-butylphenyl)-3,4,9,10-perylenecarboximide, and N,N'-bis(2,6-xylidyl)-3,4,9,10-perylenecarboximide, and wherein the reference dye provides for an oxygen-insensitive fluorescence emission signal; (b) contacting the fluid sample with the distal end portion of the optical sensor; (c) exciting the indicator composition at using radiation of a first wavelength, to produce an oxygen indicator emission signal at a second wavelength and a reference dye emission signal at a third wavelength; (d) calculating the apparent quantity of oxygen present in the fluid sample from the oxygen indicator emission signal; and (e) correcting the apparent quantity of oxygen present for variations resulting from external factors, by determining the ratio of the oxygen indicator emission signal to the reference dye emission signal.

Claim 11

A method for measuring oxygen in a fluid sample, comprising: (a) providing an optical sensor comprising an optical waveguide having a distal end portion for contacting the fluid sample, and a proximal end portion for communication with means for receiving a signal from the distal end portion, and wherein the distal end portion has an oxygen sensor means comprising a cross-linked oxygen-permeable membrane of a cured perfluorinated urethane polymer, wherein the cured perfluorinated urethane polymer is selected from the group consisting of (i) a cured perfluorinated polymer comprising a perfluorinated urethane polymer precursor, having the structural formula ##STR8## wherein Ar is a monocyclic aromatic moiety and X is a perfluorinated polyether linkage containing approximately 2 to 100 recurring perfluorinated monomer units having the structure (--CF 2 O--), (--CF 2 CF 2 O--), or combinations thereof, cross-linked with a cross-linking agent, (ii) a perfluorinated urethane polymer precursor having the structural formula ##STR9## which has been cured with a photoinitiator in the presence of a radiation, wherein n is an integer in the range of 0 to 6 inclusive, and (iii) an epoxy-terminated precursor having the structural formula ##STR10## which has been cured with a photoinitiator in the presence of radiation, and, physically entrapped therein, an indicator composition of an oxygen indicator and a reference dye, wherein the oxygen indicator provides for an oxygen-sensitive fluorescence emission signal, and wherein the reference dye provides for an oxygen-insensitive fluorescence emission signal; (b) contacting the fluid sample with the distal end portion of the optical sensor; (c) exciting the indicator composition at using radiation of a first wavelength, to produce an oxygen indicator emission signal at a second wavelength and a reference dye emission signal at a third wavelength; (d) calculating the apparent quantity of oxygen present in the fluid sample from the oxygen indicator emission signal; and (e) correcting the apparent quantity of oxygen present for variations resulting from external factors, by determining the ratio of the oxygen indicator emission signal to the reference dye emission signal.

Claim 12

A method for measuring oxygen in a fluid sample, comprising: (a) providing an optical sensor comprising an optical waveguide having a distal end portion for contacting the fluid sample, and a proximal end portion for communication with means for receiving a signal from the distal end portion, and wherein the distal end portion has an oxygen sensor means comprising a cross-linked oxygen-permeable membrane of a cured perfluorinated urethane polymer, wherein the cured perfluorinated urethane polymer is selected from the group consisting of (i) a cured perfluorinated polymer comprising a perfluorinated urethane polymer precursor, having the structural formula ##STR8## wherein Ar is a monocyclic aromatic moiety and X is a perfluorinated polyether linkage containing approximately 2 to 100 recurring perfluorinated monomer units having the structure (--CF 2 O--), (--CF 2 CF 2 O--), or combinations thereof, cross-linked with a cross-linking agent, (ii) a perfluorinated urethane polymer precursor having the structural formula ##STR9## which has been cured with a photoinitiator in the presence of a radiation, wherein n is an integer in the range of 0 to 6 inclusive, and (iii) an epoxy-terminated precursor having the structural formula ##STR10## which has been cured with a photoinitiator in the presence of radiation, and, physically entrapped therein, an indicator composition of an oxygen indicator and a reference dye, wherein the oxygen indicator provides for an oxygen-sensitive fluorescence emission signal, and wherein the reference dye provides for an oxygen-insensitive fluorescence emission signal; (b) contacting the fluid sample with the distal end portion of the optical sensor; (c) exciting the indicator composition at using radiation of a first wavelength, to produce an oxygen indicator emission signal at a second wavelength and a reference dye emission signal at a third wavelength; (d) calculating the apparent quantity of oxygen present in the fluid sample from the oxygen indicator emission signal; and (e) correcting the apparent quantity of oxygen present for variations resulting from external factors, by determining the ratio of the oxygen indicator emission signal to the reference dye emission signal.