In vivo spectral micro-imaging of tissue

• US 8,320,650 B2
• Filed: 11/09/2009
• Issued: 11/27/2012
• Est. Priority Date: 11/30/2005
• Status: Active Grant

• Current Assignee: Lawrence Livermore National Security LLC (Government of the United States of America), Regents of the University of California (University of California)
• Original Assignee: Lawrence Livermore National Security LLC (Government of the United States of America), Regents of the University of California (University of California)
• Inventors: Demos, Stavros G., Urayama, Shiro, Lin, Bevin, Saroufeem, Ramez Moussa Ghobrial
• Primary Examiner(s): STREGE, JOHN B
• Application Number:
  US12/615,085
• Publication Number:
  US 20100134605A1
• Time in Patent Office: 1,114 Days
• Field of Search: 382/128, 356/51, 600/473
• US Class Current: 382/128
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• CPC Class Codes: A61B 5/0084 for introduction into the body, e.g. by catheters A61B5/1459 takes precedence
• CPC Class Codes: A61B 5/0088 for oral or dental tissue
• CPC Class Codes: A61B 5/0091 for mammography
• CPC Class Codes: A61B 5/411 Detecting or monitoring allergy or intolerance reactions to an allergenic agent or substance
• CPC Class Codes: A61B 5/417 the bone marrow
• CPC Class Codes: G01N 2021/4792 Polarisation of scatter light
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G02B 21/082 for incident illumination only
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 23/24 Instruments or systems for viewing the inside of hollow bodies, e.g. fibrescopes

Abstract

In vivo endoscopic methods an apparatuses for implementation of fluorescence and autofluorescence microscopy, with and without the use of exogenous agents, effectively (with resolution sufficient to image nuclei) visualize and categorize various abnormal tissue forms.

Claims

1. A method, comprising:

illuminating target tissue with ultraviolet (UV) light having a wavelength of less than 400 nm to produce fluorescense;

providing imaging optics capable of producing microscopic resolution within a range from about 0.2 μ

m to about 5 μ

m, wherein a portion of said flourescense is directed toward said imaging optics to produce directed flourescense;

optically collecting, with said imaging optics, substantially all of said directed fluorescense to produce collected fluorescense; and

producing an image from said collected fluorescense;

wherein said image comprises an image spatial resolution, wherein said target tissue is within a living organism, wherein the step of optically collecting substantially all of said directed fluorescense is accomplished within a time duration that is no greater than the time it takes said target tissue to move, with respect to said imaging optics, the distance of said image spatial resolution.

2. The method of claim 1, wherein said fluorescence comprises autofluorescense (AF).

3. The method of claim 1, further comprising visually displaying said image.

4. The method of claim 1, wherein said UV light is selected to illuminate said target tissue to a depth consisting essentially of about 100 μ

m or less.

5. The method of claim 1, wherein said imaging optics comprise a configuration selected from the group consisting of a wide-field microscope and wide-field microendoscope.

6. The method of claim 1, wherein the step of optically collecting substantially all of said directed fluorescense is accomplished within a time duration of about 200 μ

s or less.

7. The method of claim 1, wherein said image comprises a maximum image spatial resolution, wherein said target tissue is within a living organism, wherein the step of optically collecting substantially all of said directed fluorescense is accomplished within a time duration that is no greater than the time it takes said target tissue to move, with respect to said imaging optics, the distance of a desired said spatial resolution of said image, wherein said desired said spatial resolution is no greater than said maximum image spatial resolution.

8. The method of claim 1, wherein said UV light comprises a wavelength selected from the group consisting of a wavelength within a range from about 180 nm to about 300 nm, a wavelength within a range from about 300 nm to about 320 nm, a wavelength within a range from about from about 320 nm to about 400 nm.

9. The method of claim 1, wherein the step of illuminating target tissue is accomplished with a source of said UV light, wherein said UV light is coupled from said source into a fiber optic, wherein said fiber optic is operatively fixed within an endoscope, wherein said imaging optics are operatively fixed within said endoscope and wherein said imaging optics have an image plane thickness of less than about 100 μ

m.

10. The method of claim 1, wherein said target tissue is selected from the group consisting of any tissue that is accessible by an endoscope within a living organism, any tissue within a living organism, any tissue within a living human being, cancer tissue, stomach tissue, oral tissue, kidney tissue, colon tissue, breast tissue, skin tissue, nail tissue, brain tissue, pancreas tissue, bile duct tissue, bladder tissue, esophagus tissue, bone marrow tissue, intestine tissue, oral tissue and lung tissue.

11. The method of claim 2, wherein exogenous agent has been contacted to said target tissue.

12. The method of claim 2, wherein no exogenous agent has been contacted to said target tissue.

13. The method of claim 1, wherein the step of producing an image from said directed fluorescense is accomplished without the use of a spatial filter.

14. The method of claim 1, wherein the step of optically collecting substantially all of said directed fluorescense to produce collected fluorescense includes collecting said directed fluorescense for an exposure time of about or less than a desired image resolution divided by the relative velocity of said target tissue with respect to said imaging optics.

15. The method of claim 2, further comprising taking a biopsy of said target tissue when said image shows suspect tissue selected from the group consisting of normal columnar mucosa having a honeycomb like pattern with goblet cells, a glandular crypt which appears as one or more depressions or large dark holes on the surface of columnar epithelium, dysplastic epithelium where the surface is includes multiple papillary, villiform or cauliform structures, disorganized tissue with loss of the specific cellular outline patterns characteristic of normal squamous or columnar epithelia and epithelial mucosa with abnormal blood vascular pattern or angiogenesis associated with malignant neoplasia.

16. The method of claim 2, further comprising therapeutically treating said target tissue when said image shows suspect tissue selected from the group consisting of normal columnar mucosa having a honeycomb like pattern with goblet cells, a glandular crypt which appears as one or more depressions or large dark holes on the surface of columnar epithelium, dysplastic epithelium where the surface is includes multiple papillary, villiform or cauliform structures, disorganized tissue with loss of the specific cellular outline patterns characteristic of normal squamous or columnar epithelia and epithelial mucosa with abnormal blood vascular pattern or angiogenesis associated with malignant neoplasia.

17. The method of claim 2, further comprising identifying said target tissue as suspected diseased tissue when said image shows a tissue image selected from the group consisting of normal columnar mucosa having a honeycomb like pattern with goblet cells, a glandular crypt which appears as one or more depressions or large dark holes on the surface of columnar epithelium, dysplastic epithelium where the surface is includes multiple papillary, villiform or cauliform structures, disorganized tissue with loss of the specific cellular outline patterns characteristic of normal squamous or columnar epithelia and epithelial mucosa with abnormal blood vascular pattern or angiogenesis associated with malignant neoplasia.

18. The method of claim 1, wherein said collected flourescense substantially comprises said directed flourescense less the amount of said directed flourescense attenuated by said imaging optics.

19. The method of claim 1, wherein said UV light comprises at least two wavelengths, wherein the step of optically collecting substantially all of said directed fluorescense is accomplished within a time duration of about 200 μ

s or less.

20. An apparatus, comprising:

means for illuminating target tissue with ultraviolet (UV) light having a wavelength of less than 400 nm to produce fluorescense;

imaging optics capable of producing microscopic resolution within a range from about 0.2 μ

m to about 5 μ

m, wherein said imaging optics are fixed such that a portion of said flourescense is directed toward said imaging optics to produce directed flourescense, wherein said imaging optics substantially collect all of said directed fluorescense to produce collected fluorescense; and

means for producing an image from said collected fluorescense;

wherein said imaging optics produce an image spatial resolution that is no less than the time it takes said target tissue to move, with respect to said imaging optics, the distance of said image spatial resolution.

21. The apparatus of claim 20, wherein said fluorescence comprises autofluorescense (AF).

22. The apparatus of claim 20, wherein said UV light can be selected to illuminate said target tissue to a depth consisting essentially of about 100 μ

m or less.

23. The apparatus of claim 20, wherein said imaging optics comprise a configuration selected from the group consisting of a wide-field microscope and wide-field microendoscope.

24. The apparatus of claim 20, wherein said imaging optics substantially collect all of said directed fluorescense within a time duration of about 200 μ

s or less.

25. The apparatus of claim 20, wherein said UV light comprises a wavelength selected from the group consisting of a wavelength within a range from about 180 nm to about 300 nm, a wavelength within a range from about 300 nm to about 320 nm, a wavelength within a range from about from about 320 nm to about 400 nm.

26. The apparatus of claim 20, wherein said means for illuminating target tissue is accomplished with a source of said UV light, said apparatus further comprising a fiber optic, wherein said UV light is coupled from said source into said fiber optic, said apparatus further comprising an endoscope, wherein said fiber optic is operatively fixed within said endoscope, wherein said imaging optics are operatively fixed within said endoscope and wherein said imaging optics have an image plane thickness of less than about 100 μ

m.

27. The apparatus of claim 20, wherein said means for producing an image from said directed fluorescense is accomplished without the use of a spatial filter.