Device for measuring light-activated fluorescence and its use

• US 20060175555A1
• Filed: 06/02/2005
• Published: 08/10/2006
• Est. Priority Date: 12/14/1999
• Status: Active Grant

• Current Assignee: Uwe Kirschner
• Original Assignee: Uwe Kirschner
• Inventors: Lau, Matthias
• Granted Patent: US 7,417,238 B2
• Time in Patent Office: Days
• US Class Current: 250/486.100
• CPC Class Codes: G01J 1/0425 using optical fibers
• CPC Class Codes: G01J 1/58 using luminescence generated by light
• CPC Class Codes: G01J 3/02 Details
• CPC Class Codes: G01J 3/0256 Compact construction
• CPC Class Codes: G01J 3/0272 Handheld
• CPC Class Codes: G01J 3/0286 Constructional arrangements for compensating for fluctuations caused by temperature, humidity or pressure, or using cooling or temperature stabilization of parts of the device; Controlling the atmosphere inside a spectrometer, e.g. vacuum
• CPC Class Codes: G01J 3/0291 Housings; Spectrometer accessories; Spatial arrangement of elements, e.g. folded path arrangements
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01N 2021/6432 Quenching
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 2201/08 Optical fibres; light guides

Abstract

The invention relates to a device for measuring light-activated fluorescence of at least one coating that contains a fluorescent material, and its use for measuring fluid materials which cause fluorescence-quenching in at least one of the fluorescent coatings. To activate the fluorescence, at least one first light wave-guide is directed onto at least one coating applied to a support and the fluorescent light is directed at a detector by means of at least one-second light wave-guide, in order to determine the intensity of the fluorescent light. The end faces of the different fluorescent light wave guides are then arranged to have overlapping entry and/or exit cones and/or be of a shape substantially identical to the at least one coating containing a fluorescent material, in such a way that an accurate measurement of the fluorescence intensity can be attained, and that the light source(s), light wave guides and the detector(s) are lodged in a measuring head.

Claims

1. A device for measuring fluorescence excited by light comprising a measuring head, the measuring head comprising:

a) an upper measuring head region having a face designed to have a fluorescing material applied thereto, or be in contact with a fluorescing material, or with a support having fluorescing material applied thereto;

b) at least one light source which emits light of at least one wavelength which excites at least one fluorescence, and thus, fluorescent light, in the at least one layer;

c) at least one first optical conductor which directs the light from the at least one light source directly into or in the direction of the fluorescing material;

d) at least one detector for measuring the intensity of the fluorescent light; and

e) at least one second optical conductor receiving the fluorescent light generated in the fluorescing layer, the at least one second optical conductor directing the fluorescent light onto the at least one detector;

wherein the end faces of the at least one first optical conductor and the at least one second optical conductor are arranged relative to one another as a function of their numerical apertures to achieve accurate measured values of the intensity of the fluorescent light by having the entry and/or exit cone of the said at least one optical conductor overlap the entry and/or exit cone of the at least one second optical conductor.

2. The device defined in claim 1, comprising the at least one first optical conductor and the at least one second optical conductor arranged as a bundle in the shape of a ring with the at least one first optical conductor arranged in the interior of the ring.

3. The device defined in claim 1, comprising the at least one first optical conductor and the at least one second optical conductor arranged along a linearly extending axis.

4. The device defined in claim 1, comprising:

a) at least one third optical conductor receiving the reflected light generated by the fluorescing layer; and

b) at least one detector for measuring the intensity of the reflected light.

5. Device according to claim 1, wherein a part of the measuring head holds the outer ends of the first and the second optical conductors, and at least the part of the measuring head which holds the outer ends of the first and the second optical conductors is of flexible construction.

6. Device according to claim 1, wherein the measuring head has an upper region, which is at least partially bent.

7. Device according to claim 1, wherein at least one of a filter, a system of exchangeable filters, or a launching optical system is arranged between the light source and at least one first optical conductor.

8. Device according to claim 1, wherein at least one of the second optical conductors receiving the fluorescent light and the first optical conductors transmitting the exciting light from the light source are arranged in the shape of a ring, a circular arc or a star on an end of the measuring head, wherein at least some of the first optical conductors have an axis which is not parallel to the axis of at least some of the second optical conductors, with all of the first and the second optical conductors pointing towards the at least one layer containing fluorescing material.

9. Device according to claim 6, wherein at least one of the second optical conductors is arranged in an inner ring or first and second optical conductors are arranged in form of a ring with an alternating arrangement.

10. Device according to claim 1, wherein the at least one first and the at least one second optical conductors are inclined at different angles with their ends pointing towards the fluorescing layer.

11. Device according to claim 1, wherein there is arranged on an upper measuring head region a heater having a temperature sensor and a controller or regulator which is arranged in the measuring head and maintains a prescribable temperature at the fluorescing layer(s).

12. Device according to claim 1, wherein the support, which is transparent to exciting light and fluorescent light, has a surface which contains partially polished or reflecting surface regions or is surrounded by a medium of lower refractive index, and is mounted in an exchangeable fashion on the measuring head.

13. Device according claim 10, wherein exciting light is launched into the support with the aid of at least one optical conductor such that the exciting light is totally reflected at least in the region of the layer, and attenuated total reflection occurs.

14. Device according to claim 10, wherein the support is constructed in an elongated fashion in a plane.

15. Device according to claim 10, wherein the support is subdivided along its longitudinal axis into a plurality of regions.

16. Device according to claim 10, wherein, on an end face opposite and end face into which the exciting light can be launched, the support has an angular surface and a layer of the at least one layer which contains fluorescing material and at which the exciting and fluorescing light is reflected in the direction of a planar optical conductor constructed symmetrically relative to the support, and the light from the angular surface thereof is directed onto an end face arranged at the other end of an optical conductor, and from there at least fluorescent light is directed onto a detector via at least one of the optical conductors, the support and planar optical conductor being arranged at a spacing from one another or being optically separated into the region of the angular surface.

17. Device according to claim 10, wherein the support is of unshaped construction comprising two limbs, the two limbs are optically separated from one another, and the exciting light can be launched into an end face of a limb via at least one additional optical conductor, and at least fluorescent light can be coupled out via the end face of the other limb into at least one further optical conductor, which at least one additional optical conductor and at least one further optical conductor are in addition to the at least one first and at least one second optical conductors.

18. Device according to claim 15, wherein the two limbs of the u-shaped support are connected in the shape of a bow, a wedge or a cone, or by means of an angular web.

19. Device according to claim 1, wherein at least one of the heating elements or temperature sensor elements are integrated into the support.

20. Device according to claim 1, wherein between one of the optical conductors and one of the at least one layers containing the fluorescing material, a transparent body made from optically scattering material is arranged or a body comprising a diffusely scattering surface is positioned facing the layer.

21. Device according to claim 18, wherein the body is formed from optically transparent material which contains light-scattering particles or a material in which a wavelength specific absorption occurs.

22. Device according to claim 1, wherein at least one further optical conductor directs reference light onto a further detector for detecting a reference signal.

23. Device according to claim 1, wherein an upper heated region is thermally insulated with respect to a lower region, in which lower region the light source(s) and the detector(s) are held.

24. Device according to claim 1, wherein said device is configured to detect fluorescence-quenching, fluid materials.

25. Device according to claim 1, wherein the support is configured to receive heating elements.

26. A device for measuring fluorescence excited by light comprising:

a) a support;

b) at least one layer of a material applied to the support, the at least one layer of material having a planar fluorescing layer containing a fluorescing material;

c) at least one light source which emits light of at least one wavelength which excites at least one fluorescence, and thus, fluorescent light, in the at least one layer;

d) at least one first optical conductor which directs the light from the at least one light source directly into or in the direction of the at least one planar layer of material;

e) at least one detector for measuring the intensity of the fluorescent light;

f) at least one second optical conductor receiving the fluorescent light generated in the at least one layer, the at least one second optical conductor directing the fluorescent light onto the at least one detector; and

g) a measuring head holding the at least one first optical conductor, the at least one second optical conductor, and the at least one light source;

wherein the end faces of the at least one first optical conductor and the at least one second optical conductor are arranged relative to one another as a function of their numerical apertures and as a function of the at least one planar layer of material to achieve accurate local assignment of the measured values of the intensity of the fluorescent light, and wherein the at least one first optical conductor and the at least one second optical conductor are arranged as a bundle in the shape of a ring with the at least one first optical conductor arranged in the interior of the ring.

27. Device according to claim 26, wherein a part of the measuring head holds the outer ends of the optical conductors, and at least the part of the measuring head which holds the outer ends of the optical conductors is of flexible construction.

28. Device according to claim 26, wherein the measuring head has an upper region, which is at least partially bent.

29. Device according to claim 26, wherein at least one of a filter, a system of exchangeable filters or a launching optical system is arranged between the light source and at least one first optical conductor.

30. Device according to claim 26, wherein at least one of the optical conductors consist of conductors receiving the fluorescent light and optical conductors transmitting the exciting light from the light source are arranged in the shape of a ring, a circular arc or a star on an end of the measuring head pointing independent to each other towards the at least one layer containing fluorescing material.

31. Device according to claim 26, wherein at least one of the second optical conductors is arranged in an inner ring or first and second optical conductors are arranged in form of a ring with an alternating arrangement.

32. Device according to claim 26, wherein the at least one first and the at least one second optical conductors are inclined at different angles with their ends pointing towards the fluorescing layer.

33. Device according to claim 26, wherein there is arranged on an upper measuring head region a heater having a temperature sensor and a controller or regulator which is arranged in the measuring head and maintains a prescribable temperature at the fluorescing layer(s).

34. Device according to claim 26, wherein the support, which is transparent to exciting light and fluorescent light, has a surface which contains partially polished or reflecting surface regions or is surrounded by a medium of lower refractive index, and is mounted in an exchangeable fashion on the measuring head.

35. Device according claim 32, wherein exciting light is launched into the support with the aid of at least one optical conductor such that the exciting light is totally reflected at least in the region of the layer, and attenuated total reflection occurs.

36. Device according to claim 32, wherein the support is constructed in an elongated fashion in a plane.

37. Device according to claim 32, wherein the support is subdivided along its longitudinal axis into a plurality of regions.

38. Device according to claim 32, wherein, on an end face opposite an end face into which the exciting light can be launched, the support has an angular surface and a layer of the at least one layer which contains fluorescing material and at which the exciting and fluorescing light is reflected in the direction of a planar optical conductor constructed symmetrically relative to the support, and the light from the angular surface thereof is directed onto an end face arranged at the other end of an optical conductor, and from there at least fluorescent light is directed onto a detector via at least one of the optical conductors, the support and planar optical conductor being arranged at a spacing from one another or being optically separated into the region of the angular surface.

39. Device according to claim 32, wherein the support is of u-shaped construction comprising two limbs, the two limbs are optically separated from one another, and the exciting light can be launched into an end face of a limb via at least one additional optical conductor, and at least fluorescent light can be coupled out via the end face of the other limb into at least one further optical conductor, which at least one additional optical conductor and at least one further optical conductor are in addition to the at least one first and at least one second optical conductors.

40. Device according to claim 37, wherein the two limbs of the u-shaped support are connected in the shape of a bow, a wedge or a cone, or by means of an angular web.

41. Device according to claim 32, wherein at least one of heating elements or temperature sensor elements are integrated into the support.

42. Device according to claim 32, wherein between one of the optical conductors and one of the at least one layers containing the fluorescing material, a transparent body made from optically scattering material is arranged or a body comprising a diffusely scattering surface is positioned facing the layer.

43. Device according to claim 40, wherein the body is formed from optically transparent material, which contains light-scattering particles, or a material in which a wavelength specific absorption occurs.

44. Device according to claim 32, wherein at least one further optical conductor directs reference light onto a further detector for detecting a reference signal.

45. Device according to claim 32, wherein an upper heated region is thermally insulated with respect to a lower region, in which lower region the light source(s) and the detector(s) are held.

46. Device according to claim 32, wherein said device is configured to detect fluorescence-quenching, fluid materials.

47. Device according to claim 32, wherein the support is configured to receive heating elements.