Diagnostic fluorescence and reflectance

• US 20040064053A1
• Filed: 09/30/2002
• Published: 04/01/2004
• Est. Priority Date: 09/30/2002
• Status: Abandoned Application

• Current Assignee: BC Cancer Agency, Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: BC Cancer Agency
• Inventors: Cox, Dennis, Malpica, Anais, Chang, Sung K., Staerkel, Gregg, Mirabal, Yvette, Atkinson, E. Neely, Richards-Kortum, Rebecca, Follen, Michele, Utzinger, Urs, MacAulay, Calum
• Application Number:
  US10/262,610
• Publication Number:
  US 20040064053A1
• Time in Patent Office: Days
• US Class Current: 600/478
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: A61B 5/0075 by spectroscopy, i.e. measuring spectra, e.g. Raman spectroscopy, infrared absorption spectroscopy A61B5/0071 takes precedence
• CPC Class Codes: A61B 5/0084 for introduction into the body, e.g. by catheters A61B5/1459 takes precedence
• CPC Class Codes: A61B 5/4331 of the cervix A61B5/435 takes precedence
• CPC Class Codes: G01N 2021/1734 Sequential different kinds of measurements; Combining two or more methods
• CPC Class Codes: G01N 2021/174 either absorption-reflection or emission-fluorescence
• CPC Class Codes: G01N 2021/4745 Fused bundle, i.e. for backscatter
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6423 Spectral mapping, video display
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 21/474 Details of optical heads therefor, e.g. using optical fibres
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

Systems and methods are described for improved diagnostic fluorescence and reflectance. A method of detecting tissue abnormality in a tissue sample in vivo detects a set of reflectance spectra emitted from a tissue sample as a result of illumination with an excitation light from a fiber optic probe that has at least one collection fiber positioned at a source-detector separation, and determining if the tissue sample is normal or abnormal based on the resulting reflectance spectra. Another method of detecting tissue abnormality in a tissue sample in vivo includes illuminating the tissue sample in vivo with at least one electromagnetic radiation wavelength selected to cause the tissue sample to produce a set of fluorescence intensity spectra indicative of tissue abnormality, detecting the resulting fluorescence intensity spectra, and determining if the tissue sample is normal or abnormal based on the resulting fluorescence intensity spectra. Yet another method of detecting tissue abnormality in a tissue sample in vivo includes combining the two methods described above.

Claims

1. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with excitation light from a fiber optic probe;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation; and

determining from the set of reflectance spectra whether the tissue sample is normal or abnormal.

2. The method of claim 1, wherein the calculating step comprises pre-processing the set of reflectance spectra to reduce patient-to-patient variation.

3. The method of claim 1, wherein the calculating step comprises conducting principal component analysis of the reflectance spectra.

4. The method of claim 1, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

5. The method of claim 4, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

6. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with a first and second electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-350 nm and the second electromagnetic wavelength being selected from the range 370-450 nm;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra whether the tissue sample is normal or abnormal.

7. The method of claim 6, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

8. The method of claim 6, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra to reduce patient-to-patient variations.

9. The method of claim 6, wherein the calculating step comprises conducting principal component analysis of the fluorescence spectra.

10. The method of claim 6, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

11. The method of claim 10, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

12. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with a first and second electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-340 nm and the second electromagnetic wavelength being selected from the range 410-420 nm;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

de from the set of fluorescence intensity spectra whether the tissue sample is normal or abnormal.

13. The method of claim 12, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

14. The method of claim 12, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra to reduce patient-to-patient variations.

15. The method of claim 12, wherein the calculating step comprises conducting principal component analysis of the fluorescence spectra.

16. The method of claim 12, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

17. The method of claim 16, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

18. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with a first and second electromagnetic wavelenth, the first electromagnetic wavelength being selected from the range 330-350 nm and the second electromagnetic wavelength being selected from the range 400-450 nm;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra whether the tissue sample is normal or abnormal.

19. The method of claim 18, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

20. The method of claim 18, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra to reduce patient-to-patient variations.

21. The method of claim 18, wherein the calculating step comprises conducting principal component analysis of the fluorescence spectra.

22. The method of claim 18, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

23. The method of claim 22, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

24. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with a single electromagnetic wavelength, the single electromagnetic wavelength being selected from the range 370-400 nm;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra whether the tissue sample is normal or abnormal.

25. The method of claim 24, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

26. The method of claim 24, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra to reduce patient-to-patient variations.

27. The method of claim 24, wherein the calculating step comprises conducting principal component analysis of the fluorescence spectra.

28. The method of claim 24, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

29. The method of claim 28, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

30. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with a first, second, and third electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-340 nm, the second electromagnetic wavelength being selected from the range 350-380 nm, and the third electromagnetic wavelength being selected from the range 400-450 nm;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra whether the tissue sample is normal or abnormal.

31. The method of claim 30, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

32. The method of claim 30, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra to reduce patient-to-patient variations.

33. The method of claim 30, wherein the calculating step comprises conducting principal component analysis of the fluorescence spectra.

34. The method of claim 30, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

35. The method of claim 34, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

36. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first, second, and third electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-360 nm, the second electromagnetic wavelength being selected from the range 420-430 nm, and the third electromagnetic wavelength being selected from the range 460-470 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

37. The method of claim 36, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

38. The method of claim 36, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

39. The method of claim 36, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

40. The method of claim 36, wherein the calculating step comprises conducting principal component analysis of the set of fluorescence intensity spectra and the set of reflectance spectra.

41. The method of claim 36, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

42. The method of claim 41, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

43. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first and second electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-360 nm, and the second electromagnetic wavelength being 460 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

44. The method of claim 43, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

45. The method of claim 43, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

46. The method of claim 43, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

47. The method of claim 43, wherein the calculating step comprises conducting principal component analysis of the fluorescence intensity spectra and the set of reflectance spectra.

48. The method of claim 43, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

49. The method of claim 48, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

50. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first and second set of electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-350 nm and the second electromagnetic wavelength being 470 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

51. The method of claim 50, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

52. The method of claim 50, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

53. The method of claim 50, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

54. The method of claim 50, wherein the calculating step comprises conducting principal component analysis of the fluorescence intensity spectra and the set of reflectance spectra.

55. The method of claim 50, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

56. The method of claim 55, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

57. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first and second electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-350 nm and the second electromagnetic wavelength being selected from the range 470-480 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

58. The method of claim 57, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

59. The method of claim 57, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

60. The method of claim 57, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

61. The method of claim 57, wherein the calculating step comprises conducting principal component analysis of the fluorescence intensity spectra and the set of reflectance spectra.

62. The method of claim 57, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

63. The method of claim 62, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

64. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first, second, and third electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 350-360 nm, the second electromagnetic wavelength being selected from the range 420-430 nm, and the third electromagnetic wavelength being 460 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least one collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the-tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

65. The method of claim 64, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

66. The method of claim 64, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

67. The method of claim 64, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

68. The method of claim 64, wherein the calculating step comprises conducting principal component analysis of the fluorescence intensity spectra and the set of reflectance spectra.

69. The method of claim 64, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

70. The method of claim 69, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.

71. A method of detecting tissue abnormality in a tissue sample in vivo comprising:

providing a tissue sample;

sequentially illuminating the tissue sample in vivo with an excitation light and a first and second electromagnetic wavelength, the first electromagnetic wavelength being selected from the range 330-350 nm, the second electromagnetic wavelength being selected from the range 460-470 nm;

detecting, with the fiber optic probe, a set of reflectance spectra emitted from the tissue sample as a result of illumination with the excitation light, the fiber optic probe comprising at least two collection fiber positioned at at least one source-detector separation selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation;

detecting the set of fluorescence intensity spectra emitted from the tissue sample as a result of illumination; and

determining from the set of fluorescence intensity spectra or the set of reflectance spectra, or a combination of the set of fluorescence intensity spectra and the set of reflectance spectra whether the tissue sample is normal or abnormal.

72. The method of claim 71, wherein the at least one source-detector separation is selected from the group consisting of 250 μ

m separation, 1.1 mm separation, 2.1 mm separation, and 3.0 mm separation.

73. The method of claim 71, wherein the calculating step comprises truncating the set of fluorescence intensity spectra at 700 nm.

74. The method of claim 71, wherein the calculating step comprises pre-processing the set of fluorescence intensity spectra and the set of reflectance spectra to reduce patient-to-patient variations.

75. The method of claim 71, wherein the calculating step comprises conducting principal component analysis of the fluorescence intensity spectra and the set of reflectance spectra.

76. The method of claim 71, wherein the calculating step comprises selecting and classifying the tissue sample using Mahalanobis distance.

77. The method of claim 76, wherein the calculating step further comprises cross-validating results from selecting and classifying the tissue sample using Mahalanobis distance.