Optical measurement apparatus and method therefor

• US 5,811,312 A
• Filed: 08/30/1995
• Issued: 09/22/1998
• Est. Priority Date: 08/31/1993
• Status: Expired due to Fees

• Current Assignee: Nissui Pharmaceutical Co. Ltd. (Shimadzu Corporation)
• Original Assignee: Daikin Industries Ltd
• Inventors: Ueda, Tomoaki, Yoshida, Masakazu, Masuda, Kenji, Sakamoto, Tomomi, Hasegawa, Masanori, Shigemori, Kazuhisa
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/428,120
• Time in Patent Office: 1,119 Days
• Field of Search: 385/12, 385/129, 385/130, 356/317, 356/318, 422/55, 422/58, 422/82.05, 422/82.08, 422/82.11, 435/287.1, 435/287.2, 435/288.7, 435/808, 436/527, 436/535, 436/164, 436/165, 436/172, 436/805, 436/807
• US Class Current: 436/527
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• 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 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures

Abstract

An optical measurement apparatus includes a reaction vessel 2 which is formed in one body with a slab-type optical waveguide 1, and the apparatus adds fine-grains or water soluble dye for absorbing a fluorescent light which is radiated from fluorescent dye, and/or an exciting light so that stray light due to reagent is reduced and the S/N ratio of the optical measurement is improved.

Claims

1. An optical measurement apparatus comprising:

a reaction vessel for holding liquids;

an optical waveguide for propagating light therewithin, the optical waveguide forming at least one surface of the reaction vessel wherein a substance is fixed on the surface for binding to a substance to be measured;

means for radiating an exciting light to the optical waveguide at a first predetermined angle relative to the optical waveguide;

a fluorescent substance which produces fluorescent light when excited by the exciting light contained within the reaction vessel for labeling at least a portion of a substance to be measured which is bound to the substance fixed on the surface of the waveguide;

a light absorbing material contained within the reaction vessel, the light absorbing material absorbing light at a wavelength of the exciting light or at a wavelength of the fluorescent light; and

means for measuring fluorescent light which is emitted from the fluorescent substance labeling a portion of a substance to be measured which is bound to the substance fixed on the surface of the waveguide at a second predetermined angle relative to the optical waveguide.

2. An optical measurement apparatus as recited in claim 1 wherein the first predetermined angle and the second predetermined angle are the same.

3. An optical measurement apparatus as recited in claim 1, wherein the light absorbing material is a water soluble dye.

4. An optical measurement apparatus as recited in claim 1, wherein the light absorbing material is fine grains of material.

5. An optical measurement apparatus as recited in claim 4, wherein the light absorbing material is fine grains of material selected from the group consisting of carbon black, metal, polystyrene and polyethylene.

6. An optical measurement apparatus as recited in claim 5, wherein the light absorbing material is fine grains of platinum or gold.

7. An optical measurement apparatus as recited in claim 4, wherein the fine grains of material have a diameter of about 200 nm.

8. An optical measurement apparatus as recited in claim 4, wherein a concentration of the fine grains of material in the reaction vessel ranges from 0.004%-0.3%.

9. An optical measurement apparatus as recited in claim 1, wherein the fluorescent light propagates within the waveguide in a totally reflective manner.

10. An optical measurement apparatus as recited in claim 1, wherein each of the fluorescently labeled substance, the substance to be measured and the substance fixed to the surface of the optical waveguide is a ligand.

11. The optical measurement method as recited in claim 10, wherein each of the fluorescently labeled substance, the substance to be measured and the substance fixed to the surface of the optical waveguide is an antigen or an antibody.

12. An optical measurement apparatus as recited in claim 1, wherein the light absorbing material absorbs light at a wavelength of the exciting light and at a wavelength of the fluorescent light.

13. An optical measurement apparatus as recited in claim 1, wherein the first and second predetermined angles are parallel to a length of the waveguide.

14. An optical measurement apparatus as recited in claim 1, wherein the first predetermined angle is perpendicular to a length of the waveguide, and the second predetermined angle is parallel to the length of the waveguide.

15. An optical measurement apparatus as recited in claim 1, wherein the exciting light is transmitted to the waveguide through a collimator and a neutral density filter.

16. An optical concentration measurement method comprising;

pouring a test liquid containing a substance to be measured into a reaction vessel and pouring a reagent liquid containing a fluorescent substance, which fluorescent substance produces fluorescent light when excited by an exciting light, into the reaction vessel, such that at least a portion of the substance to be measured binds to a substance fixed to a surface of an optical waveguide which forms a wall of the reaction vessel, and at least a portion of the fluorescent substance binds to the at least a portion of the substance to be measured bound to the substance fixed to the surface of the optical waveguide, so as to constrain the at least a portion of the substance to be measured and the at least a portion of the fluorescent substance in a vicinity of the optical waveguide,

adding a light absorbing material to the reaction vessel, which light absorbing material absorbs light at an exciting wavelength of the fluorescent substance or at an emitting wavelength of the fluorescent substance,

introducing an exciting light into the optical waveguide at a first predetermined angle relative to the optical waveguide for exciting the at least a portion of the fluorescent substance constrained in the vicinity of the surface of the optical waveguide, whereby the at least a portion of the fluorescent substance constrained in the vicinity of the surface of the optical waveguide is excited by the exciting light into emitting fluorescent light,

measuring the emitted fluorescent light which is output from the waveguide at a second predetermined angle relative to the optical waveguide,

determining the concentration of the substance to be measured from the measurement of the emitted fluorescent light.

17. An optical measurement methods as recited in claim 16, wherein the first predetermined angle and the second predetermined angle are the same.

18. The optical measurement method as recited in claim 17, wherein the first and second predetermined angles are parallel to a length of the waveguide.

19. The optical measurement method as recited in claim 16, wherein the light absorbing material is a water soluble dye.

20. The optical measurement method as recited in claim 16, wherein the light absorbing material is fine grains of material.

21. The optical measurement method as recited in claim 20, wherein the light absorbing material is fine grains of material selected from the group consisting of carbon black, metal, polystyrene and polyethylene.

22. The optical measurement method as recited in claim 21, wherein the light absorbing material is fine grains of platinum or gold.

23. The optical measurement method as recited in claim 20, wherein the fine grains of material have a diameter of about 200 nm.

24. The optical measurement method as recited in claim 20, wherein a concentration of the fine grains of material in the reaction vessel ranges from 0.004%-0.3%.

25. The optical measurement method as recited in claim 16, wherein the fluorescent light propagates within the waveguide in a totally reflective manner.

26. The optical measurement method as recited in claim 16, wherein the light absorbing material absorbs light at a wavelength of the exciting light and at a wavelength of the fluorescent light.

27. The optical measurement method as recited in claim 16, wherein the exciting light is transmitted to the waveguide through a collimator and a neutral density filter.

28. The optical measurement method as recited in claim 16, wherein the first predetermined angle is perpendicular to a length of the waveguide, and the second predetermined angle is parallel to the length of the waveguide.

29. The optical measurement method as recited in claim 16, wherein each of the fluorescently labeled substance, the substance to be measured and the substance fixed to the surface of the optical waveguide is a ligand.

30. The optical measurement method as recited in claim 29, wherein each of the fluorescently labeled substance, the substance to be measured and the substance fixed to the surface of the optical waveguide is an antigen or an antibody.

31. An optical concentration measurement method comprising:

providing a substance fixed to a surface of an optical waveguide which forms a wall of a reaction vessel,

pouring a test liquid containing a substance to be measured into the reaction vessel, the substance to be measured being labeled with a fluorescent label which produces fluorescent light when excited by an exciting light, such that at least a portion of the substance to be measured binds to the substance fixed to the surface of the optical waveguide so as to constrain the at least a portion of the substance to be measured to the in a vicinity of the optical waveguide,

adding a light absorbing material to the reaction vessel, which light absorbing material absorbs light at an exciting wavelength of the fluorescent label or at an emitting wavelength of the fluorescent label,

introducing an exciting light into the optical waveguide at a first predetermined angle relative to the optical waveguide, for exciting the fluorescent label of the at least a portion of the substance to be measured constrained in the vicinity of the surface of the optical waveguide, whereby the fluorescent label of the at least a portion of the substance to be measured constrained at the surface of the optical waveguide is excited by the exciting light into emitting fluorescent light,

measuring the emitted fluorescent light which is output from the waveguide at a second predetermined angle relative to the optical waveguide,

determining the concentration of the substance to be measured from the measurement of the emitted fluorescent light.

32. The optical measurement method as recited in claim 31, wherein the light absorbing material is a water soluble dye.

33. The optical measurement method as recited in claim 31, wherein the light absorbing material is fine grains of material.

34. The optical measurement method as recited in claim 33, wherein the fine grains of material have a diameter of about 200 nm.

35. The optical measurement method as recited in claim 33, wherein a concentration of the fine grains of material in the reaction vessel ranges from 0.004%-0.3%.

36. The optical measurement method as recited in claim 31, wherein the light absorbing material is fine grains of material selected from the group consisting of carbon black, metal, polystyrene and polyethylene.

37. The optical measurement method as recited in claim 36, wherein the light absorbing material is fine grains of platinum or gold.

38. The optical measurement method as recited in claim 31, wherein the light absorbing material absorbs light at a wavelength of the exciting light and at a wavelength of the fluorescent light.

39. The optical measurement method as recited in claim 31, wherein the fluorescent light propagates within the waveguide in a totally reflective manner.

40. An optical measurement method as recited in claim 31, wherein the first predetermined angle and the second predetermined angle are the same.

41. The optical measurement method as recited in claim 40, wherein the first and second predetermined angles are parallel to a length of the waveguide.

42. The optical measurement method as recited in claim 31, wherein the first predetermined angle is perpendicular to a length of the waveguide, and the second predetermined angle is parallel to the length of the waveguide.

43. The optical measurement method as recited in claim 31, wherein the exciting light is transmitted to the waveguide through a collimator and a neutral density filter.

44. The optical measurement method as recited in claim 31, wherein each of the substance to be measured and the substance fixed to the surface of the optical waveguide is a ligand.

45. The optical measurement method as recited in claim 44, wherein each of the substance to be measured and the substance fixed to the surface of the optical waveguide is an antigen or an antibody.