TRI-COLOR DUAL GLUCOSE AND OXYGEN SENSORS AND METHODS OF PREPARING AND USING THEM

• US 20140273043A1
• Filed: 03/13/2014
• Published: 09/18/2014
• Est. Priority Date: 03/14/2013
• Status: Active Grant

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Meldrum, Deirdre, Tian, Yanqing, Zhang, Liqiang, Su, Fengyu, Buizer, Sean
• Granted Patent: US 10,156,573 B2
• Time in Patent Office: Days
• US Class Current: 435/14
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/66 involving blood sugars, e.g. galactose
• CPC Class Codes: Y10T 436/144444 Glucose
• CPC Class Codes: Y10T 436/207497 Molecular oxygen

Abstract

The present disclosure relates to an optical fluorescence sensor comprising a probe for sensing glucose, an intra-reference probe, and a matrix. The present disclosure also relates to an optical fluorescence dual sensor comprising a probe for sensing glucose, a probe for sensing oxygen, an intra-reference probe, and a matrix. The present disclosure additionally relates to methods of preparing these sensors and methods of using them.

Claims

1. An optical fluorescence sensor comprising:

(a) a probe for sensing glucose having formula I;

2. The optical fluorescence sensor according to claim 1, wherein the probe for sensing glucose is:

3. The optical fluorescence sensor according to claim 1, wherein the intra-reference probe is:

8. A method of preparing an optical fluorescence sensor comprising the steps of:

(a) copolymerizing an intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide onto a substrate; and

(b) attaching or immobilizing a probe for sensing glucose onto the substrate;

wherein the probe for sensing glucose and the intra-reference probe are as defined in claim 1.

9. A method of preparing an optical fluorescence sensor comprising the step of:

copolymerizing a probe for sensing glucose and an intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide onto a substrate;

wherein the probe for sensing glucose is as defined in claim 1; and

the intra-reference probe is defined in claim 1.

12. The method according to claim 8, wherein copolymerizing the intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide forms poly(2-hydroxyethyl methacrylate), poly(N-(6-aminohexyl)methacrylamide), poly(acrylamide), poly(2-hydroxyethyl methacrylate)-co-polyacrylamide-co-poly(N-(6-aminohexyl)methacrylamide) and their composites with the polymerized probes.

13. The method according to claim 9, wherein copolymerizing the probe for sensing glucose and the intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide forms poly(2-hydroxyethyl methacrylate), poly(N-(6-aminohexyl)methacrylamide), poly(acrylamide), poly(2-hydroxyethyl methacrylate)-co-polyacrylamide-co-poly(N-(6-aminohexyl)methacrylamide) and their composites with the polymerized probes.

18. A method of determining the concentration of glucose in a sample comprising:

(a) exposing the sample to an optical fluorescence sensor according to claim 1 or an optical fluorescence dual sensor according to claim 4;

(b) irradiating the sensor at a first wavelength to produce a glucose indicator emission signal at a second wavelength and an intra-reference emission signal at a third wavelength;

(c) measuring the glucose indicator emission signal at the second wavelength;

(d) measuring the intra-reference emission signal at the third emission wavelength; and

(e) ratiometrically determining the concentration of glucose in the sample.

21. The method according to claim 18, wherein more than one sample is used.

22. The method according to claim 18, wherein the sample comprises a bodily fluid.

23. The method according to claim 22, wherein the bodily fluid comprises blood.

24. The method according claim 18, wherein the sample is obtained from a cell culture or a subject.

25. The method according to claim 18, wherein the sample is selected from the group consisting of live single cells, live several cells, live cell clusters, and live tissue.

4. An optical fluorescence dual sensor comprising:

(a) a probe for sensing glucose having formula I;

5. The optical fluorescence dual sensor according to claim 4, wherein the probe for sensing glucose is:

6. The optical fluorescence dual sensor according to claim 4, wherein the probe for sensing oxygen is:

7. The optical fluorescence dual sensor according to claim 4, wherein the intra-reference probe is:

10. A method of preparing an optical fluorescence dual sensor comprising the steps of:

(a) copolymerizing a probe for sensing oxygen and an intra-reference probe, with poly(2-hydroxyethyl methacrylate), polyacrylamide, and poly(2-hydroxyethyl methacrylate)-co-polyacrylamide onto a substrate; and

(b) attaching or immobilizing a probe for sensing glucose onto the substrate;

wherein the probe for sensing glucose, the probe for sensing oxygen and the intra-reference probe are defined in claim 4.

11. A method of preparing an optical fluorescence dual sensor comprising the step of:

copolymerizing a probe for sensing glucose, a probe for sensing oxygen and an intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide onto a substrate;

wherein the probe for sensing glucose is as defined in claim 4; and

the probe for sensing oxygen and the intra-reference probe are as defined in claim 4.

14. The method according to claim 11, wherein copolymerizing the probe for sensing glucose, the probe for sensing oxygen and the intra-reference probe, with 2-hydroxyethyl methacrylate, N-(6-aminohexyl)methacrylamide, and acrylamide forms poly(2-hydroxyethyl methacrylate), poly(N-(6-aminohexyl)methacrylamide), poly(acrylamide), poly(2-hydroxyethyl methacrylate)-co-polyacrylamide-co-poly(N-(6-aminohexyl)methacrylamide) and their composites with the polymerized probes.

15. The method according to claim 8 or 10, wherein the probe for sensing glucose is:

16. The method according to claim 10 or 11, wherein the probe for sensing oxygen is:

17. The method according to any one of claims 8 to 11, wherein the intra-reference probe is:

19. A method of determining the concentration of oxygen in a sample comprising:

(a) exposing the sample to an optical fluorescence dual sensor according to claim 4;

(b) irradiating the sensor at a first wavelength to produce an oxygen indicator emission signal at a second wavelength and an intra-reference emission signal at a third wavelength;

(c) measuring the oxygen indicator emission signal at the second wavelength;

(d) measuring the intra-reference emission signal at the third wavelength; and

(e) ratiometrically determining the oxygen concentration in the sample.

20. A method of simultaneously determining the concentration of glucose and the concentration of oxygen in a sample comprising

(a) exposing the sample to an optical fluorescence dual sensor according to claim 4;

(b) irradiating the sensor at a first wavelength to produce a glucose indicator emission signal at a second wavelength, an oxygen indicator emission signal at a third wavelength and an intra-reference emission signal at a fourth wavelength;

(c) measuring the glucose indicator emission signal at the second wavelength;

(d) measuring the oxygen indicator emission signal at the third wavelength;

(e) measuring the intra-reference emission signal at the fourth wavelength;

(f) ratiometrically determining the concentration of glucose in the sample using the measurements obtained in steps (c) and (e); and

(g) ratiometrically determining the oxygen concentration in the sample using the measurements obtained in steps (d) and (e).