Flourescence reader

• US 20070231883A1
• Filed: 03/22/2007
• Published: 10/04/2007
• Est. Priority Date: 03/22/2006
• Status: Active Grant

• Current Assignee: Crimson International Assets LLC (The Carlyle Group LP (Corporate Private Equity))
• Original Assignee: Åmic AB (Johnson & Johnson)
• Inventors: Mendel-Hartvig, Ib, Vilhelmsson, Kennet, Ohman, Ove, Lindstrom, Tomas, Backlund, Johan
• Granted Patent: US 8,691,153 B2
• Time in Patent Office: Days
• US Class Current: 435/287.200
• CPC Class Codes: G01N 2021/6465 Angular discrimination
• 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

Abstract

Fluorescence reader (10) for an optical assay arrangement comprising a polymeric sample substrate (1) having a reaction site-surface and a substrate surface (3), the fluorescence reader comprising a light source (5) arranged to illuminate the reaction site-surface through the substrate surface, and a detector device (6) arranged to detect fluorescent light emitted from said reaction site-surface and transmitted through the substrate surface, the substrate surface provided with total-internal-reflection suppressing means (15).

Claims

1. A fluorescence reader (10) for a sample substrate (1) comprising:

a reaction site-surface (2);

a substrate surface (3) opposed to said reaction site-surface (2);

a light source (5) positioned to inject exciting light rays into the substrate surface (3); and

a detector device (6) positioned to detect a fluorescent light emitted from the reaction site-surface (2) and transmitted through the substrate surface (3), wherein said substrate surface (3) is provided with a total-internal-reflection-suppressing means (12, 15, 16) located in an optical path of the emitted fluorescent light to increase transmission through the substrate surface (3) for detection by the detector device (6).

2. A fluorescence reader according to claim 1, wherein said total-internal-reflection-suppressing means (12, 15, 16) is arranged to release emitted fluorescent light trapped within the sample substrate (1) for detection by the detector device (6).

3. A fluorescence reader according to claim 1 wherein said light source (5) is arranged to inject exciting light rays into the substrate surface (3) with an incidence angle (7) substantially coinciding with the maximum emission angle (8) of the emitted fluorescent light wherein the incident angle is measured relative to the normal of the reaction site surface.

4. A fluorescence reader according to claim 3, wherein said substrate surface (3) is provided with an incidence-angle-controlling means (11) located in the optical path of the exciting light rays.

5. A fluorescence reader according to claim 4, wherein said incidence-angle-controlling means (11) comprises a surface relief structured entry-section (9) designed to admit exciting light into the sample substrate (1) with the incidence angle (7), wherein said incidence angle is enlarged and wherein the incident angle is measured relative to the normal of the reaction site surface.

6. A fluorescence reader according to claim 5, wherein said surface relief structured entry-section (11) comprises a diffractive structure.

7. A fluorescence reader according to claim 5, wherein said surface relief structured entry-section (11) comprises a refractive structure.

8. A fluorescence reader according to claim 2 wherein said total-internal-reflection-suppressing means (12, 15, 16) comprises a surface relief structured exit-section (12) designed to diffract or refract the emitted fluorescent light out of the substrate.

9. A fluorescence reader according to claim 8, wherein said surface relief structured exit-section (12) is designed to focus the emitted fluorescent light.

10. A fluorescence reader according to claim 8 wherein said surface relief structured-exit section (12) comprises a diffractive structure.

11. A fluorescence reader according to claim 8 said surface relief-structured exit section (12) comprises a refractive structure.

12. A fluorescence reader according to claim 8, wherein the design of a surface relief structure is arranged to vary over a surface of the surface relief structured exit-section (12) in correspondence with the varying emission angle (8) of impinging fluorescent light.

13. A fluorescence reader according to claim 8, wherein the position and extension of the surface relief structured exit-section (12) determines the position and extension of an analyzed reaction site area.

14. A fluorescence reader according to claim 1, wherein a light collecting lens device (13) is positioned to receive the emitted fluorescent light transmitted through the substrate surface (3).

15. A fluorescence reader according to claim 8, wherein a light-collecting body (15) is located in close proximity to the surface relief structured exit-section (12) provided on said substrate surface (3).

16. A fluorescence reader according to claim 1, wherein said total-internal-reflection-suppressing means comprises a light-collecting body (15) positioned in optically wetting contact with said substrate surface (3).

17. A fluorescence reader according to claim 16, wherein said light-collecting (15) body is designed to collect and transmit light by means of total internal-reflection.

18. A fluorescence reader according to claim 16, wherein said light-collecting body (15) is substantially ellipsoid-shaped.

19. A fluorescence reader according to claim 16, wherein said light-collecting body (15) is provided with an input port (14) for the exciting light rays.

20. A fluorescence reader according to claim 16, wherein said light-collecting body (15) is provided with at least one output port.

21. A fluorescence reader according to claim 16, wherein said light-collecting body (15) has a refractive index that substantially corresponds to, or is larger than, the refractive index of the sample substrate (1).

22. A fluorescence reader according to claim 1, wherein said total-internal-reflection-suppressing means (12, 15, 16) comprises an optically wetting layer (16) having a suitable refractive index.

23. A fluorescence reader according to claim 22, wherein at least a portion of said optically wetting layer (16) is attached to a lens device (17) arranged to focus the emitted fluorescent light on the detector device (6).

24. A fluorescence reader according claim 23, wherein said optically wetting layer (16) has a refractive index that is higher than the refractive index of the sample substrate (1), and lower than the refractive index of the lens device (17).

25. A fluorescence reader according to claim 22, wherein at least a portion of said optically wetting layer (16) is attached to the detector device (6).

26. A fluorescence reader according to claim 4, wherein said incidence-angle-controlling means (11) comprises an optically wetting layer (16) having a suitable refractive index.

27. A fluorescence reader according to claim 26, wherein said optically wetting layer (16) comprises a soft polymeric material.

28. A fluorescence reader according to claim 1, wherein said detector device (6) is provided with spectral filtering means arranged to prevent detection of the wavelengths of the exciting light rays.

29. A fluorescence reader according to claim 1, wherein said detector device (6) is provided with polarization filtering means.

30. A fluorescence reader according to claim 1, wherein said light source (5) is provided with spectral filtering means arranged to prevent transmission of the wavelengths coinciding with the emitted fluorescent light.

31. A fluorescence reader according to claim 1, wherein excitation and measurement of the emitted fluorescent light is performed in different geometrical planes.

33. A sample substrate (1) comprising:

a reaction site-surface (2); and

a substrate surface (3) opposed to said reaction site-surface (2), wherein said sample substrate (1) is adapted for the fluorescence reader of claim 1.

34. A sample substrate according to claim 33, wherein said sample substrate (1) is made of a polymeric material.

35. A sample substrate according to claim 33, wherein said reaction site-surface (2) is arranged to form lines and/or spots of fluorophores.

36. A sample substrate according to claim 33, wherein the reaction site surface (2) is provided with a pattern of protruding microstructures.

37. A sample substrate according to claim 36, wherein said protruding microstructures comprise micro posts enabling a capillary flow.

38. A sample substrate according to claim 33, wherein the substrate surface (3) is provided with a surface relief structured exit section (12) configured to suppress the total internal-reflection of emitted fluorescence light rays.

39. A sample substrate according to claim 33, wherein the substrate surface (3) is provided with a surface relief structured entry section (11) configured to enlarge an incidence angle (7) of the exciting light rays.

40. A sample substrate according to claim 39, wherein the surface relief structured entry-section (11) is a diffractive structure.

41. A sample substrate according to claim 39, wherein the surface relief structured entry-section (11) is a refractive structure.

32. A method of detecting the fluorescence of a sample substrate using a fluorescence reader comprising:

a) injecting exciting light rays into a substrate site surface with a light source positioned to inject exciting light rays into the substrate surface wherein said substrate surface is opposed to a reaction site-surface and

b) detecting fluorescent light emitted from the reaction-site surface with a detector device positioned to detect the fluorescent light emitted from the reaction site-surface and transmitted through the substrate surface wherein said substrate surface is provided with a total-internal-reflection-suppressing means located in an optical path of the fluorescent light to increase transmission through the substrate surface for detection by the detector device wherein the fluorescence reader is positioned in an optical assay arrangement.