Method and apparatus for noninvasive measurement of carotenoids and related chemical substances in biological tissue

• US 6,205,354 B1
• Filed: 06/18/1999
• Issued: 03/20/2001
• Est. Priority Date: 06/18/1999
• Status: Expired due to Term

• Current Assignee: Utah State University
• Original Assignee: University of Utah (State of Utah)
• Inventors: Gellermann, Werner, Bernstein, Paul S., McClane, Robert W., Katz, Nikita B.
• Primary Examiner(s): Casler, Brian L.
• Application Number:
  US09/335,932
• Time in Patent Office: 641 Days
• Field of Search: 600/473, 600/476, 600/475, 600/477, 600/310, 600/318, 356/301, 356/303, 606/2-4, 606/10
• US Class Current: 600/477
• CPC Class Codes: A61B 5/14546 for measuring analytes not otherwise provided for, e.g. ions, cytochromes
• CPC Class Codes: A61B 5/1455 using optical sensors, e.g. spectral photometrical oximeters
• CPC Class Codes: A61B 5/443 Evaluating skin constituents, e.g. elastin, melanin, water
• CPC Class Codes: G01N 2021/656 Raman microprobe
• CPC Class Codes: G01N 21/65 Raman scattering
• CPC Class Codes: G01N 21/658 enhancement Raman, e.g. surface plasmons

Abstract

A method and apparatus are provided for the determination of levels of carotenoids and similar chemical compounds in biological tissue such as living skin. The method and apparatus provide a noninvasive, rapid, accurate, and safe determination of carotenoid levels which in turn can provide diagnostic information regarding cancer risk, or can be a marker for conditions where carotenoids or other antioxidant compounds may provide diagnostic information. Such early diagnostic information allows for the possibility of preventative intervention. The method and apparatus utilize the technique of resonance Raman spectroscopy to measure the levels of carotenoids and similar substances in tissue. In this technique, laser light is directed upon the area of tissue which is of interest. A small fraction of the scattered light is scattered inelastically, producing the carotenoid Raman signal which is at a different frequency than the incident laser light, and the Raman signal is collected, filtered, and measured. The resulting Raman signal can be analyzed such that the background fluorescence signal is subtracted and the results displayed and compared with known calibration standards.

Claims

1. A method for noninvasively determining the presence or risk of a malignancy disease in biological tissue, comprising the steps of:

obtaining a light sourse which generates light at a wavelength that produces a Raman tenoids to be detected;

directing light from the light sourse onto biological tissue for which carotenoid levels are to be measured, the light having an intensity which does not cause destruction of the tissue and does not substantially alter carotenoid levels in the tissue;

collecting light scattered from the tissue, the scattered light including elastically and inelastically scattered light, the inelastically scattered light producing a Raman signal corresponding to carotenoid in the tissue;

filtering out the elastically scattered light; and

quantifying the intensity of the Raman signal, wherein a substantial difference between the intensity of the Raman signal and the intensity of Raman scattering from adjkacent normal biological tissue indicates the presence or risk of a malignancy disease.

2. The method of claim 1, wherein the light source generates light in a wavelength which overlaps the absorption bands of the carotenoids to be detected.

3. The method of claim 1, wherein the light source generates laser light in a wavelength range from about 450 nm to about 520 nm.

4. The method of claim 1, wherein the biological tissue resides in a live subject.

5. The method of claim 1, wherein the biological tissue is living skin.

6. The method of claim 5, wherein the light has an intensity of up to about 200 mW/cm².

7. The method of claim 5, wherein the scattered light is measured at frequencies characteristic of carotenoids in the skin.

8. The method of claim 1, wherein the Raman signal is quantified via signal intensity calibrated with actual carotenoid levels.

9. A method for noninvasively determining the antioxidant status in skin tissue, comprising the steps of:

obtaining a light source which generates light at a wavelength that produces a Raman response with a wavelength shift for carotenoids to be detected in skin tissue;

directing light from the light source onto skin tissue for which carotenoid levels are to be measured, the light having an intensity which does not cause destruction of the tissue and does not substantially alter carotenoid levels in the tissue;

collecting light scattered from the tissue, the scattered light including elastically and inelastically scattered light, the inelastically scattered light producing a Raman signal corresponding to carotenoids in the tissue;

filtering out the elastically scattered light; and

quantifying the intensity of the Raman signal in order to assess the antioxidant status of the tissue.

10. The method of claim 9, wherein the light source generates light in a wavelength which overlaps the absorption bands of the carotenoids to be detected.

11. The method of claim 9, wherein the light source generates laser light in a wavelength range from about 450 nm to about 520 nm.

12. The method of claim 9, wherein the skin tissue resides in a live subject.

13. The method of claim 9, wherein the skin tissue is living skin.

14. The method of claim 13, wherein the scattered light is measured at frequencies characteristic of carotenoids in the skin.

15. The method of claim 9, wherein the Raman signal is quantified via signal intensity calibrated with actual carotenoid levels.

16. A method for noninvasive measurement of carotenoids in skin tissue, comprising the steps of:

obtaining a light source which generates light at a wavelength that produces a Raman response with a wavelength shift for carotenoids to be detected;

directing light from the light source onto skin tissue for which carotenoid levels are to be measured, the light having an intensity which does not cause destruction of the skin tissue and does not substantially alter carotenoid levels in the skin tissue;

collecting light scattered from the skin tissue, the scattered light including elastically and inelastically scattered light, the inelastically scattered light producing a Raman signal corresponding to carotenoids in the skin tissue;

filtering out the elastically scattered light;

quantifying the intensity of the Raman signal; and

subtracting a background fluorescence signal of the skin tissue from the Raman signal of the carotenoids being detected.

17. The method of claim 16, wherein the light source generates light in a wavelength which overlaps the absorption bands of the carotenoids to be detected.

18. The method of claim 16, wherein the light source generates laser light in a wavelength range from about 450 nm to about 520 nm.

19. The method of claim 16, wherein the light has an intensity of up to about 200 mW/cm².

20. The method of claim 16, wherein the scattered light is measured at frequencies characteristic of carotenoids in the skin.

21. The method of claim 16, wherein the Raman signal is quantified via signal intensity calibrated with actual carotenoid levels.

22. An apparatus for noninvasive measurement of carotenoids and related chemical substances in biological tissue, comprising:

a laser light source that generates light at a wavelength giving a Raman response with a wavelength shift for carotenoids being detected;

a light delivery and collection module for directing light onto biological tissue and collecting scattered light from the tissue such that the light does not damage the tissue or substantially alter the carotenoid levels in the tissue, the light delivery and collection module comprising;

a first lens for collimating laser light;

a first narrow bandwidth filter in optical communication with the first lens;

first and second dichroic beam splitters in optical communication with the first narrow bandwidth filter;

a second lens adapted to direct a beam of laser light from the second dichroic beam splitter to tissue and collect scattered light from the tissue;

a second narrow bandwidth filter in optical communication with the second lens; and

a third lens in optical communication with the second narrow bandwidth filter;

a spectrally selective system for selecting Raman shifted light from the collected scattered light;

detection means for scanning and measuring the Raman shifted light at frequencies characteristic of carotenoids; and

quantifying means for determining Raman signal intensity for the carotenoids being detected.

23. The apparatus of claim 22, wherein the light source generates laser light in a wavelength range from about 450 nm to about 520 nm.

24. The apparatus of claim 22, wherein the spectrally selective system is a grating spectrometer.

25. The apparatus of claim 22, wherein the spectrally selective system includes a grating monochromator.

26. The apparatus of claim 22, wherein the spectrally selective system includes a holographic filter.

27. The apparatus of claim 22, wherein the spectrally selective system includes a dielectric filter.

28. The apparatus of claim 22, wherein the spectrally selective system includes an acousto-optic filter.

29. The apparatus of claim 22, wherein the detection means comprises a CCD detector array.

30. The apparatus of claim 22, wherein the detection means comprises an intensified CCD detector array.

31. The apparatus of claim 22, wherein the quantifying means comprises a personal computer.

32. The apparatus of claim 22, wherein the quantifying means comprises a CCD image display or monitor.