Fluorometric detection using visible light

• US 6,914,250 B2
• Filed: 12/06/2002
• Issued: 07/05/2005
• Est. Priority Date: 03/07/1997
• Status: Expired due to Term

• Current Assignee: Clare Chemical Research Incorporated
• Original Assignee: Clare Chemical Research Incorporated
• Inventors: Seville, Mark
• Primary Examiner(s): Hannaher, Constantine
• Application Number:
  US10/313,892
• Publication Number:
  US 20030173525A1
• Time in Patent Office: 942 Days
• Field of Search: 250/458.1, 250/459.1
• US Class Current: 250/458.1
• 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 21/6447 by visual observation

Abstract

Systems, devices and methods of the present invention are for viewing a pattern of fluorophors capable of fluorescing when exposed to visible light, e.g., fluorescently stained DNA, protein or other biological material. The system includes a light source emitting light in the visible spectrum, such as a fluorescent lamp used in domestic lighting, a first optical filter capable of transmitting light from the source at wavelengths capable of exciting the fluorophors and of absorbing light of other wavelengths, and a second optical filter capable of blocking substantially all the light from the source not blocked by the first filter, so that the only light reaching the viewer is light produced by fluorescence of the fluorophors.

Claims

1. A visible light photoluminescent imaging system for viewing one or more patterns of fluorescence emitted by fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said system comprising:

a) an electroluminescent light-producing element having a luminous surface that produces light;

b) a first optical filter positioned between said the luminous surface of the electroluminescent light-producing element and said fluorophors, wherein the first optical filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type; and

c) a second optical filter positioned in optical communication with said fluorophors, wherein said second optical filter is capable of transmitting light of said emitted type from said fluorophors and of substantially preventing transmission of said excitation light;

said system being constructed and arranged such that patterns of emission from the fluorophors are viewable.

2. The visible light photoluminescent imaging system of claim 1 wherein the electroluminescent light-producing element comprises one or more liquid crystals.

3. The visible light photoluminescent imaging system of claim 2 wherein the electroluminescent light-producing element comprises a liquid crystal display panel.

4. The visible light photoluminescent imaging system of claim 3 wherein the electroluminescent light-producing element comprises a computer monitor.

5. The visible light photoluminescent imaging system of claim 3 wherein the liquid crystal display panel has a white color setting.

6. The visible light photoluminescent imaging system of claim 3 wherein the liquid crystal display panel has a blue color setting.

7. The visible light photoluminescent imaging system of claim 3 wherein the liquid crystal display panel has a blue-green color setting.

8. The visible light photoluminescent imaging system of claim 3 wherein the liquid crystal display panel has a green color setting.

9. The visible light photoluminescent imaging system of claim 1 wherein the electroluminescent light-producing element has a power consumption of less than or equal to about 24 W.

10. The visible light photoluminescent imaging system of claim 1 comprising at least one additional optical filter positioned in optical communication with said second optical filter, wherein the additional optical filter is capable of transmitting light of said emitted type form said fluorophors and of substantially preventing transmission of infrared light.

11. The visible light photoluminescent imaging system of claim 1 comprising a detector in optical communication with the second optical filter.

12. The visible light photoluminescent imaging system of claim 11 wherein the detector is positioned at an angle 45°

or more from normal incidence with respect to a plane containing the second filter.

13. The visible light photoluminescent imaging system of claim 1 wherein the electroluminescent light-producing element has an illumination surface that has a non-uniform luminous flux across the illumination surface.

14. The visible light photoluminescent imaging system of claim 1 wherein the electroluminescent light-producing element has an illumination surface that has a substantially uniform luminous flux across the illumination surface.

15. The visible light photoluminescent imaging system of claim 1 wherein the wavelength of light produced by the electroluminescent imaging system is uniform across the illumination surface.

16. The visible light photoluminescent imaging system of claim 1 wherein the wavelength of light produced by the electroluminescent imaging system is non-uniform across the illumination surface.

17. The visible light photoluminescent imaging system of claim 1 comprising a computer operationally connected to the electroluminescent light-producing element, wherein the computer controls the light produced by the electroluminescent light-producing element.

18. The visible light photoluminescent imaging system of claim 17 wherein the computer selects the wavelengths of the light produced by the electroluminescent light-producing element.

19. The visible light photoluminescent imaging system of claim 17 wherein the computer selects the spatial distribution of light across the illumination surface.

20. The visible light photoluminescent imaging system of claim 17 wherein the computer controls the intensity of the light produced by the electroluminescent light-producing element.

21. The visible light photoluminescent imaging system of claim 1 wherein the electroluminescent light-producing element comprises one or more light emitting diodes.

22. The visible light photoluminescent imaging system of claim 1 wherein said first optical filter and said second optical filter are integrated into a gel cassette.

23. The visible light photoluminescent imaging system of claim 1 wherein said fluorophors are provided in a gel cassette.

24. A field monitoring device for viewing one or more patterns of emission from fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said system comprising:

a) an excitation source comprising at least one light-producing element that produces light, at least a portion of which is capable of exciting said fluorophors, and a first optical filter, positioned between said light-producing element and said fluorophors that is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type, wherein the excitation source produces light substantially free of light in the ultraviolet region; and

b) a second filter positioned in optical communication with said fluorophors, wherein said second optical filter is capable of transmitting light of said emitted type from said fluorophors and of substantially preventing transmission of light from said excitation source;

said system being constructed and arranged such that patterns of emission from said fluorophors are viewable.

25. The photoluminescent field monitoring device of claim 24 comprising a detector.

26. The photoluminescent field monitoring device of claim 24 wherein the excitation source is handheld.

27. The photoluminescent field monitoring device of claim 24 wherein the excitation source is stationary.

28. The photoluminescent field monitoring device of claim 24 comprising an underwater photoluminescent imaging system.

29. The photoluminescent field monitoring device of claim 24 wherein the excitation source has a power consumption less than about 9 W.

30. The photoluminescent field monitoring device of claim 24 operationally coupled to a microfluidic or micro-array device.

31. A method of generating a pattern of radiant emission from fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said method comprising the steps:

a) passing light of said excitation type and substantially free of ultraviolet light from an excitation source on to the fluorophors whereby the fluorophors emit light of the emitted type, wherein the excitation source comprises a light-producing element and a first optical filter positioned between said light-producing element and the fluorophors, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type; and

b) passing said emitted light through a second optical filter, which is capable of transmitting light of said emitted type from said fluorophors and capable of substantially prevent transmission of light of said excitation type.

32. The method of claim 31 wherein the fluorophors are trace contaminants in a field sample.

33. The method of claim 31 wherein the fluorophors are comprised in a genetically modified organism.

34. The method of claim 33 comprising a method wherein said fluorophors are contained in genetically-modified plants.

35. The method of claim 31 wherein the fluorophors are comprised in a tissue sample or sample of bodily fluid.

54. The method of claim 35 wherein said first optical filter and said second optical filter are integrated into a gel cassette.

36. A method of viewing a pattern of radiant emission from fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said method comprising the steps:

a) passing light of said excitation type and substantially free of ultraviolet light from an excitation source on to the fluorophors, whereby the fluorophors emit light of the emitted type, wherein the excitation source comprises a light-producing element and a first optical filter positioned between said light-producing element and the fluorophors, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type;

b) passing said emitted light through a second optical filter, which is capable of transmitting light of said emitted type from said fluorophors and capable of substantially prevent transmission of light of said excitation type; and

c) detecting said emitted light with a detector.

37. The method of claim 36 wherein the detector is a human eye.

38. The method of claim 36 wherein the detector is a charged coupled device.

39. The method of claim 36 wherein the detector is a CMOS detector.

40. The method of claim 36 wherein the detector is a camera.

41. A vessel for viewing one or more patterns of fluorescence emitted by fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said vessel comprising:

a) an excitation source comprising at least one light-producing element that produces light, at least a portion of which is capable of exciting said fluorophors, and a excitation filter positioned between said light-producing element and said fluorophors, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type;

b) a bowl capable of positioning the fluorophors in optical communication with the light source; and

c) at least one emission filter positioned in optical communication with said fluorophors, wherein said second optical filter is capable of transmitting light of said emitted type from said fluorophors and of substantially preventing transmission of light from said excitation source;

said vessel being constructed and arranged such that patterns of emission from said fluorophors are viewable.

42. The vessel of claim 41 which is a photoluminescent fish tank.

43. The vessel of claim 41 which is a photoluminescent drinking glass.

44. The vessel of claim 41 which is a photoluminescent display cabinet.

45. A method of generating one or more radiant images corresponding to images printed with ink containing fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, comprising the steps of:

a) passing light of said excitation type from an excitation source on to the fluorophors, whereby ink containing fluorophors emits light of the emitted type, wherein the excitation source comprises at least one light-producing element and a first optical filter positioned between said light-producing element and the images printed with ink containing fluorophors, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type, wherein the excitation source produces light substantially free of light in the ultraviolet region;

b) passing said emitted light through a second optical filter in optical communication with the images printed with ink containing fluorophors, wherein the second filter is capable of transmitting light of said emitted type from said fluorophors and capable of substantially preventing transmission of light of said excitation type;

whereby one or more radiant images corresponding to images printed with the ink containing fluorophors are viewable.

46. A room installation for generating radiant images, patterns, or both from fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said room installation comprising:

a) an excitation source comprising at least one light-producing element that produces light and a first optical filter positioned between said light-producing element and said fluorophors, wherein the first optical filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type, wherein the excitation source produces light substantially free of light in the ultraviolet region; and

b) a second filter positioned in optical communication with said fluorophors, wherein said second optical filter is capable of transmitting light of said emitted type from said fluorophors and of substantially preventing transmission of said excitation light.

47. The photoluminescent room installation of claim 46 wherein said excitation source is stationary.

48. The photoluminescent room installation of claim 46 wherein said excitation source is mobile.

49. The photoluminescent room installation of claim 46 wherein said second filter is incorporated into eye glasses or contact lenses.

50. A method of generating a pattern of radiant emission from an oligonucleotide sample, polynucleotide sample, or both containing fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said method comprising the steps:

a) passing light of said excitation type generated by an excitation source onto the oligonucleotide sample, polynucleotide sample, or bpth, whereby the fluorophors emit light of the emitted type, wherein the excitation source comprises a light-producing element and a first optical filter positioned between said light-producing element and the oligonucleotide sample, polynucleotide sample, or both, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type; and

b) passing said emitted light through a second optical filter, which is capable of transmitting light of said emitted type from said fluorophors and capable of substantially prevent transmission of light of said excitation type;

wherein the excitation source is configured and arranged such that the oligonucleotide sample, polynucleotide sample, or both, undergo no substantial damage over an illumination period of about five minutes.

51. The method of claim 50 wherein the oligonucleotide sample, polynucleotide sample or both contain DNA.

52. The method of claim 50 wherein said first optical filter and said second optical filter are integrated into a gel cassette.

53. A method of generating a pattern of radiant emission from fluorophors capable of being excited by light of an excitation type and capable of emitting light of an emitted type, said method comprising the steps:

a) passing light of said excitation type generated by an excitation source onto the fluorophors, whereby the fluorophors emit light of the emitted type, wherein the excitation source comprises a light-producing element and a first optical filter positioned between said light-producing element and the fluorophors, wherein the first filter is capable of transmitting light of said excitation type and of substantially preventing transmission of light of said emitted type; and

b) passing said emitted light through a second optical filter, which is capable of transmitting light of said emitted type from said fluorophors and capable of substantially prevent transmission of light of said excitation type;

wherein the excitation source is configured and arranged such that the fluorophors do not undergo substantial photo-bleaching over an illumination period of about eight minutes.