Method for quantitative analysis using optical fibers

• US 4,321,057 A
• Filed: 09/20/1979
• Issued: 03/23/1982
• Est. Priority Date: 09/20/1979
• Status: Expired due to Term

• Current Assignee: Puritan Bennett Corp. (Medtronic Plc)
• Original Assignee: Richard G. Buckles
• Inventors: Buckles, Richard G.
• Primary Examiner(s): Richman, Barry
• Application Number:
  US06/076,984
• Time in Patent Office: 915 Days
• Field of Search: 422/73, 422/68, 422/55-60, 23/230 B, 23/230 R, 356/445, 356/448, 356/435, 435/7
• US Class Current: 435/7.1
• CPC Class Codes: A61B 5/1459 invasive, e.g. introduced into the body by a catheter A61B5/1464 takes precedence
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 2021/7716 in cladding
• CPC Class Codes: G01N 2021/773 Porous polymer jacket; Polymer matrix with indicator
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/643 non-biological material
• CPC Class Codes: G01N 21/7703 using reagent-clad optical fibres or optical waveguides using measurement of total internal reflection or attenuated total reflection G01N21/552; optical fibres or waveguides per se G02B
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

A method and apparatus for chemical and biochemical analyses which employ an energy-transmissive core and may employ one or more sheaths which selectively absorb, react with, and/or filter an analyte or a product of an analyte. The core is transmissive to a chosen energy carrier and it has an inlet end and an outlet end. Between these ends it has an extended length. The passage of energy through the core is modified by reason of events which occur in one or more of the sheaths or in the case where no sheath is employed, by reason of events which occur in an ambient fluid. The resulting modification of the transmitted energy is a measure of such events which in turn are a measure of the analyte. The energy may be any of several types of energy which can be transmitted through the core from end to end and which is susceptible to modification by reactions in the sheath or sheaths or ambient fluid. The energy may be electromagnetic, electrical or sonic. In the method aspect of the invention a permeable core may be used which is bare, i.e. without a sheath, and exposed directly to an environment, e.g. the air or an industrial fluid.

Claims

1. A method for the quantitative analysis of oxygen in a fluid, said method comprising:

(a) providing a fiber-sheath element including a fiber capable of transmitting electromagnetic energy in the visible and ultraviolet ranges and defining a path for the transmission of said energy, a first oxygen-permeable sheath surrounding said fiber, an oxygen-quenchable dye incorporated into the material forming said fiber or said sheath and means for reflecting said electromagnetic radiation from said sheath into said fiber;

(b) introducing ultraviolet light into one end of said fiber whereby a portion of said ultraviolet light reacts with said fluorescent dye to produce fluorescence and emission of visible light, said emission of visible light being cumulative along a length of said path; and

(c) contacting said fiber-sheath element with said fluid whereby oxygen permeates said element and quenches said fluorescence by reaction with said dye to reduce the visible light emitted from said fiber-sheath element in proportion to the amount of oxygen contained in said fluid.

2. The method of claim 1 wherein a multiplicity of said fiber-sheath elements are employed, said elements containing said oxygen-quenchable fluorescent dye in different concentrations.

3. The method of claim 1 wherein said fluorescent dye is fluoranthrene.

4. The method of claim 1 wherein said fluorescent dye is contained in said sheath.

5. The method of claim 1 wherein said fluid is a liquid.

6. The method of claim 5 wherein said liquid is blood.

11. A method, in accordance with claim 1 or 7, wherein the fluid is a liquid and wherein one end only of said fiber-sheath element is contacted with a mass of hydrophilic material containing said liquid to be analyzed in an amount in excess of that necessary to wet said element whereby capillary action carries said liquid through the sheath to uniformly wet said element.

7. A method for the quantitative analysis of an analyte in a fluid containing said analyte species and a second species which reacts with said analyte species in the presence of a catalyst, said method comprising:

(a) providing a fiber-sheath element including;

a fiber which transmits electromagnetic radiation;

a first sheath element covering said fiber and formed of a material which is permeable to at least one of said analyte species and said second species and which contains a reagent which reacts with one of said analyte species and said second species to modify an electromagnetic energy transmitting characteristic of the fiber; and

a second sheath element surrounding said first sheath element, said second sheath element being formed of a material which is permeable to both said analyte species and said second species and which contains a catalyst for causing a reaction between said analyte species and said second species;

(b) contacting said element with the fluid to be analyzed thus allowing said analyte and said second species to permeate said second sheath element and to react in the presence of said catalyst, thereby diminishing the amount of the species reacting with said reagent;

(c) transmitting electromagnetic radiation through said fiber with cumulative modification of said radiation along a length of said fiber due to the influence of said reagent; and

(d) measuring the quantity of electromagnetic radiation of at least one wavelength emitted from said fiber.

8. The method of claim 7 further comprising:

providing an additional fiber-sheath element without said catalyst in said second sheath, and

correlating the difference between the amount of said electromagnetic radiation emitted by the two fibers to the amount of analyte present in the fluid.

9. The method of claim 8 wherein ultraviolet light is introduced from the same light source into one end of each of said fibers and wherein said reagent is a fluorescent material and the emitted electromagnetic radiation which is measured is light in the visible range.

10. The method of claim 9 wherein said analyte is glucose, said second species is oxygen, said catalyst is glucose oxidase and said reagent is an oxygen-quenchable fluorescent dye.

12. A method for quantitative analysis of an antigen A1 contained in a liquid, said method comprising:

(a) providing a fiber element capable of transmitting electromagnetic energy and defining a path for the transmission of said energy, a first sheath surrounding said fiber element and containing a first complex A2 of an antigen and an energy-modifying compound, a second sheath surrounding said first sheath, said second sheath reflecting said electromagnetic radiation and being selectively permeable to said first complex when wet, and a third sheath, surrounding said second sheath, and an antibody Ab to A1 and A2 incorporated into the material forming said third sheath;

(b) contacting said fiber element with said liquid whereby said sheaths are wetted and antigen A1, if present, diffuses from the liquid into said third sheath to complex said antibody and said first complex diffuses reversibly across said second sheath into said third sheath and likewise complexes with said antibody to produce the following competing reversible reactions in said third sheath;

space="preserve" listing-type="equation">A1+Ab⃡

A1-Ab (1)

space="preserve" listing-type="equation">A2+Ab⃡

A2-Ab (2)

(c) introducing the electromagnetic radiation into one end of said fiber whereby a portion refracts into said first sheath wherein said refracted portion reacts with said energy-modifying compound to produce a modification of the transmitted radiation which is cumulative along a length of said path and which is proportional to the amount of A1 diffusing into said fiber element.

13. The method of claim 12 wherein a multiplicity of said fiber elements are employed, said fiber elements containing said complex A2 of an antigen and an energy-modifying compound in different concentrations.

14. The method of claim 12 wherein said energy-modifying compound is a fluorescent compound, wherein said electromagnetic radiation introduced into the fiber is ultraviolet light, and said modification is conversion to visible light by fluorescence.

21. A method, in accordance with claim 12, 15, 17 or 18 wherein one end only of said fiber-sheath element is contacted with a mass of a hydrophilic material containing the liquid to be analyzed in an amount in excess of that necessary to wet said element whereby capillary action carries said liquid through said coating to uniformly wet said fiber-sheath element.

15. A method for the quantitative analysis of an antigen A1 contained in a liquid, said method comprising:

(a) providing a fiber-sheath element including a fiber which transmits electromagnetic radiation, said fiber having an index of refraction n_c and being permeable to the product Y of a reaction;

##EQU3## wherein E is an enzyme, X is a high molecular weight compound and Y is a low molecular weight compound;

a first sheath surrounding said fiber and being permeable to Y but not to X;

a second sheath surrounding said first sheath and having an index of refraction n_s lower than n_c, said second sheath being formed of a material containing an antibody Ab immunobonded to a complex of an enzyme E and an antigen A2, and an excess of X, said second sheath being permeable to A1;

(b) transmitting electromagnetic radiation selected for maximum modification by Y through said fiber;

(c) wetting said second sheath with the liquid to be analyzed, whereby any antigen A1 present in the liquid diffuses into second sheath and reacts with Ab thereby displacing A2-E in proportion to the concentration of A1 in the liquid, the displaced complex A2-E catalyzing the conversion of X into Y which then diffuses into the fiber and modifies said electromagnetic radiation cumulatively along a length of said fiber.

16. The method of claim 15 wherein a plurality of said fiber-sheath elements are provided, said elements containing different amounts of X.

19. A process in accordance with claim 15, 17 or 18 wherein Y is a dye which produces said modification of said transmitted energy by absorption.

17. A method for the quantitative analysis of an antigen contained in a liquid, said method comprising:

(a) providing a fiber-sheath element including a fiber which transmits electromagnetic radiation and which is formed of a material containing an antibody Ab, for the antigen, bonded to a compound Y in the form of a complex Ab-Y, said compound Y having an energy-modifying characteristic which changes upon immunobonding of the antigen to the antibody Ab; and

a sheath, permeable to the antigen, surrounding said fiber;

(b) transmitting electromagnetic radiation selected for maximum modification by Y through said fiber;

(c) wetting said sheath with the liquid to be analyzed, whereby any antigen present in the liquid diffuses through the sheath and into the fiber to immunobond with Ab thus modifying said characteristic of Y and said electromagnetic radiation cumulatively, along a length of said fiber.

20. The method of claim 17 or 18 wherein a plurality of said fiber-sheath elements are provided, said elements containing different amounts of Y.

18. A method for the quantitative analysis of an enzyme in a liquid sample, said method comprising:

(a) providing a fiber-sheath element including a fiber which transmits electromagnetic radiation, a sheath surrounding said fiber and formed of a hydrophilic polymer and an energy modifying compound Y incorporated into and chemically bound to said polymer, said chemical bond being selectively degradable by the enzyme to be detected, said fiber being permeable to Y;

(b) transmitting electromagnetic radiation selected for maximum modification by Y through said fiber; and

(c) wetting said sheath with the liquid to be analyzed whereby any enzyme in the liquid breaks down the chemical bond between Y and the polymer, thus releasing Y for diffusion into the fiber wherein Y modifies said radiation, cumulatively, along a length of said fiber.

22. A method for the quantitative analysis of an analyte contained in a fluid, said method comprising:

(a) providing a plurality of fiber-sheath elements, each of said elements including;

(1) an electromagnetic energy transmissive fiber defining a path for transmission of electromagnetic energy, at least a portion of said fiber being formed of a material having an electromagnetic energy transmitting characteristic capable of modification by the presence of an analyte contained in the fluid;

(2) a semipermeable sheath surrounding said fiber; and

(3) a reagent which modifies said transmitted energy responsive to the presence of an analyte or which reacts with the analyte to form a product which modifies said transmitted energy, said elements containing said reagent in different amounts;

(b) contacting said fiber-sheath elements with the fluid to be analyzed;

(c) transmitting electromagnetic radiation through said fibers with cumulative modification of said radiation along a length of each of said fibers due to the influence of said reagent; and

(d) taking the output of one of said fibers as a measurement of the concentration of the analyte and rejecting the output of the other fibers.