System and methods of fluorescence, reflectance and light scattering spectroscopy for measuring tissue characteristics

• US 8,380,268 B2
• Filed: 06/24/2005
• Issued: 02/19/2013
• Est. Priority Date: 01/19/2001
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Georgakoudi, Irene, Feld, Michael S., Zhang, Qingguo, Mueller, Markus G.
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Laryea, Lawrence N
• Application Number:
  US11/166,798
• Publication Number:
  US 20050288593A1
• Time in Patent Office: 2,797 Days
• Field of Search: 128/634, 128/665, 356/432, 356/369, 356/342, 356/364, 356/367, 356/340, 600/310, 600/473, 600/476, 600/477, 600/478, 600/407, 600/178, 600/160, 600/314, 382/128, 382/190, 395/21, 395/924
• US Class Current: 600/310
• CPC Class Codes: A61B 5/0059 using light, e.g. diagnosis by transillumination, diascopy, fluorescence A61B5/0093, A61B5/1455, A61B5/02416 and A61B5/0261 take precedence
• 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 5/1459 invasive, e.g. introduced into the body by a catheter A61B5/1464 takes precedence
• CPC Class Codes: A61B 5/4331 of the cervix A61B5/435 takes precedence
• CPC Class Codes: G01N 21/47 Scattering, i.e. diffuse reflection G01N21/25, G01N21/41 take precedence G01N21/55 takes precedence
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

The present invention utilizes a plurality of spectroscopic systems and methods to measure characteristics of tissue useful in the diagnosis of disease. In a preferred embodiment, a combination of fluorescence, reflectance and light scattered spectra can be measured and processed to provide biochemical, architectural and morphological state of tissue. The methods and systems can be used particularly in the early detection of carcinoma within tissue in vivo and in vitro.

Claims

1. A method of measuring spectral data to detect cancerous tissue comprising:

inserting a fiber optic probe within an endoscope channel, the endoscope being inserted within a body cavity or lumen, the fiber optic probe having a central light delivery optical fiber and a plurality of light collection optical fibers that surround the central light delivery optical fiber, the probe collecting fluorescent light and reflected light from the tissue at a plurality of wavelengths;

illuminating tissue with light from a broadband light source optically coupled to the central light delivery optical fiber and a laser light source optically coupled to the central delivery optical fiber such that a selected region of tissue is illuminated;

detecting the collected fluorescent light and reflected light with a detector that is optically coupled to proximal ends of the collection optical fibers;

processing detected fluorescence light and detected reflected light with a data processor to form an intrinsic fluorescence spectrum and a diffuse reflectance spectrum; and

determining a characteristic of the tissue using the data processor that processes the intrinsic fluorescence spectrum and the diffuse reflectance spectrum, in combination, to detect cancerous tissue.

2. The method of claim 1 further comprising using the intrinsic fluorescence spectrum to determine one or more components of the tissue.

3. The method of claim 1 further comprising using the diffuse reflectance spectrum to determine a size of a structure within the tissue.

4. The method of claim 3 wherein the using step further comprises generating a lookup table with different sizes of scatterers.

5. The method of claim 1 further comprising measuring a periodic component of the detected light to provide a scattered light representation.

6. The method of claim 1 further comprising analyzing the diffuse reflectance spectrum and a light scattering spectrum.

7. The method of claim 6 further comprising transforming the light scattering spectrum and removing spectral components to determine size distribution of a tissue component.

8. The method of claim 1 wherein the method of measuring spectral data further comprises calibrating with a reflectance standard and a fluorescence standard.

9. The method of claim 1 further comprising processing the intrinsic fluorescence spectrum, the diffuse reflectance spectrum and a light scattering spectrum to determine a characteristic of the tissue.

10. The method of claim 1 wherein the determining step further comprises performing discriminate analysis to diagnose the tissue.

11. The method of claim 1 further comprising using the diffuse reflectance spectrum to obtain a plurality of scattering coefficient values.

12. The method of claim 1 further comprising determining a plurality of biochemical components of the tissue using the detected fluorescence.

13. The method of claim 1 further comprising determining weight coefficients from an analysis of the detected fluorescence light and the detected reflected light.

14. An apparatus for detecting cancerous tissue comprising:

a broadband light source that illuminates a region of interest in tissue to be measured with incident light;

a laser light source;

a fiber optic probe that collects reflected light and fluorescent light from a body tissue surface at a plurality of wavelengths, the probe having a central light delivery optical fiber that is optically coupled to the laser light source and the broadband light source such that a selected tissue region can be illuminated with the laser light source and the broadband light source, and a plurality of light collection optical fibers surrounding the central light delivery fiber;

a detector system optically coupled to the collection optical fibers that detects the collected light and provides fluorescence data and reflected light data at a plurality of wavelengths; and

a data processor that processes the fluorescence data and the reflected light data to provide an intrinsic fluorescence spectrum and a diffuse reflectance spectrum, the intrinsic fluorescence spectrum and the diffuse reflectance spectrum being processed in combination to identify cancerous tissue in the region of interest.

15. The apparatus of claim 14 wherein the broadband light source generates light in a range of 330-700 nm.

16. The apparatus of claim 14 wherein the fiber optic probe has an optical shield at a distal end that couples light from the broadband light source and the laser light source to the tissue.

17. The apparatus of claim 14 wherein the probe is adapted for insertion into a working channel of an endoscope.

18. The apparatus of claim 14 further comprising a fast excitation-emission matrix instrument including the laser light source that illuminates tissue at a plurality of excitation wavelengths between 337 nm and 620 nm.

19. The apparatus of claim 14 further comprising a spectrograph optically coupled to proximal ends of the collection optical fibers.

20. The apparatus of claim 14 wherein the probe comprises six collection optical fibers within a probe that is 5 mm or less in diameter.

21. A method for detecting cancerous tissue comprising:

acquiring a fluorescence spectrum with a fiber optic probe positioned to illuminate a body tissue surface, the fiber optic probe having a central light delivery optical fiber that is optically coupled at a proximal end to a laser light source and a plurality of light collection optical fibers surrounding the central delivery optical fiber;

acquiring a reflectance spectrum with the fiber optic probe, the central delivery fiber being optically coupled at the proximal end to a broadband light source;

processing the fluorescence spectrum and the reflectance spectrum with a data processor to provide an intrinsic fluorescence spectrum and a diffuse reflectance spectrum; and

processing the intrinsic fluorescence spectrum and the diffuse reflectance spectrum, in combination, with the data processor to determine a biophysical tissue characteristic and thereby identify cancerous tissue.

22. The method of claim 21 further comprising determining a size of a structure within the tissue.

23. The method of claim 22 wherein the structure is the nuclei of epithelial cells in the tissue.

24. The method of claim 22 wherein the determining step further comprises generating a look-up table with different sizes of scatterers.

25. The method of claim 21 further comprising measuring a periodic component of the acquired reflectance spectrum.

26. The method of claim 21 further comprising calibrating a detector optically coupled to the probe using a reflectance standard and a fluorescence standard.

27. The method of claim 21 further comprising using the reflectance spectrum and the fluorescence spectrum to obtain the intrinsic fluorescence spectrum.

28. The method of claim 21 further comprising acquiring and transforming a light scattering spectrum and removing spectral components to determine size distribution of a tissue component.

29. The method of claim 21 wherein the determining step further comprises performing discriminate analysis for providing a real-time diagnosis for the tissue.

30. The method of claim 21 further comprising determining a plurality of scattering coefficient values and associating the plurality of scattering coefficient values with a plurality of different tissue regions.

31. The method of claim 21 further comprising detecting light collected by the collection optical fibers with a detector that is optically coupled to proximal ends of the collection optical fibers.