Optical fluorescence dual sensors and methods of preparing and using them

• US 8,748,192 B2
• Filed: 10/24/2012
• Issued: 06/10/2014
• Est. Priority Date: 10/25/2011
• Status: Active Grant

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Tian, Yanqing, Zhang, Weiwen, Meldrum, Deirdre, Lu, Hongguang, Jin, Yuguang, Holl, Mark
• Primary Examiner(s): Hartley, Michael G
• Assistant Examiner(s): SAMALA, JAGADISHWAR RAO
• Application Number:
  US13/659,821
• Publication Number:
  US 20130102024A1
• Time in Patent Office: 594 Days
• Field of Search: 436/81, 436/136, 436/164
• US Class Current: 436/136
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/77 by observing the effect on a chemical indicator
• CPC Class Codes: G01N 31/221 for investigating pH value
• CPC Class Codes: G01N 31/225 for oxygen, e.g. including dissolved oxygen
• CPC Class Codes: Y10T 436/207497 Molecular oxygen

Abstract

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

Claims

1. An optical fluorescence dual sensor comprising:

(a) a probe for sensing pH;

(b) a probe for sensing oxygen;

(c) an intra-reference probe; and

(d) a matrix;

wherein the probe for sensing pH has formula I;

2. The optical fluorescence dual sensor of claim 1, wherein the probe for sensing pH is:

3. The optical fluorescence dual sensor according to claim 1, wherein the probe for sensing oxygen is:

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

5. A method of determining pH of a sample comprising

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

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

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

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

(e) ratiometrically determining the pH of the sample wherein the sample is obtained from a cell culture, blood, urine, tear, industry fermentor, photobioreactor, pond, river, lake or ocean.

6. A method of determining oxygen concentration in a sample comprising

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

(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 wherein the sample is obtained from a cell culture, blood, urine, tear, industry fermentor, photobioreactor, pond, river, lake or ocean.

7. The method according to claim 5 or 6, wherein the sample comprises a microorganism.

8. The method according to claim 7, wherein the microorganism is selected from the group consisting of photosynthetic algae, cyanobacteria, Escherichia coli, Bacillus subtilis, and yeast.

9. The method according to claim 5 or 6, wherein more than one sample is used.

10. A method of simultaneously determining pH and oxygen concentration in a sample

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

(b) irradiating the sensor at a first wavelength to produce a pH 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 pH 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 pH of the sample using the measurements obtained in steps (c) and (e); and

(g) ratiometrically determining the oxygen concentration of the sample using the measurements obtained in steps (d) and (e) wherein the sample is obtained from a cell culture, blood, urine, tear, industry fermentor, photobioreactor, pond, river, lake or ocean.