Dye application for confocal imaging of cellular microstructure

• US 8,620,409 B2
• Filed: 08/04/2009
• Issued: 12/31/2013
• Est. Priority Date: 08/04/2008
• Status: Active Grant

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah Research Foundation (State of Utah)
• Inventors: Bridge, John H., Hitchcock, Robert, Sachse, Frank B.
• Primary Examiner(s): Smith, Ruth S
• Application Number:
  US13/057,419
• Publication Number:
  US 20110301438A1
• Time in Patent Office: 1,610 Days
• Field of Search: 600/407, 600/425, 600/428, 600/431, 600476-478, 424/9.6, 424/9.61
• US Class Current: 600/428
• CPC Class Codes: A61B 1/0008 characterised by distal tip features
• CPC Class Codes: A61B 1/043 for fluorescence imaging
• CPC Class Codes: A61B 5/0068 Confocal scanning
• 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 5/6852 Catheters
• CPC Class Codes: A61B 5/7285 for synchronising or triggering a physiological measurement or image acquisition with a physiological event or waveform, e.g. an ECG signal A61B6/5205, A61B6/541, A61B8/5207, A61B8/543 take precedence
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: G02B 21/0028 specially adapted for specific applications, e.g. for endoscopes, ophthalmoscopes, attachments to conventional microscopes
• CPC Class Codes: G02B 21/0032 Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence

Abstract

A system and method for confocal imaging of tissue in vivo and in situ, e.g., for minimally invasive diagnosis of patients. A catheter is provided that has a dye carrier coupled to the distal end of a fiber optics bundle, which allows for the introduction of at least one fluorescent dye therein the dye carrier into a portion of the tissue of interest of a subject or patient when the dye carrier is selectively brought into contact with the portion of the tissue of interest. The resulting confocal images permit the acquisition of diagnostic information on the progression of diseases at cellular/tissue level in patients. Furthermore, a system for ECG-triggered image acquisition is provided.

Claims

1. A method for imaging of tissue, comprising:

operatively coupling a dye carrier to a distal end of a fiber optic bundle, the dye carrier comprising at least one fluorescent dye;

positioning a portion of the dye carrier in contact with a tissue region of interest of a subject such that at least a portion of the at least one fluorescent dye diffuses into the tissue region of interest; and

acquiring a confocal image of the tissue region of interest, wherein the step of acquiring the confocal image of the tissue region of interest comprises;

transmitting light into the fiber optic bundle, through the dye carrier, and then into the tissue region of interest to excite the fluorescent dye within the tissue region of interest;

receiving light emitted from the excited fluorescent dye within the tissue region of interest, wherein the light emitted from the excited fluorescent dye is emitted through the dye carrier and then into the fiber optic bundle; and

processing the received emitted light to form the confocal image of the tissue region of interest.

2. The method of claim 1, wherein the step of receiving light emitted from the excited fluorescent dye comprises communicating the emitted light to a confocal microscopic imaging system.

3. The method of claim 1, further comprising monitoring an ECG signal taken from the subject, wherein a reference point of the ECG signal taken from the subject triggers the step of acquiring a confocal image of the tissue region of interest.

4. The method of claim 3, wherein the step of monitoring the ECG signal comprises identifying a plurality of sequential reference points of the ECG signal, and wherein the step of acquiring a confocal image of the tissue region of interest is repeated at each sequential reference point of the ECG signal.

5. The method of claim 1, wherein the dye carrier comprises a light transparent matrix.

6. The method of claim 5, wherein the at least one fluorescent dye is at a predetermined concentration.

7. The method of claim 5, wherein the dye carrier comprises at least one antibody agent.

8. The method of claim 5, wherein the light transparent matrix comprises a hydro-gel, and wherein the hydro-gel is configured to permit diffusion of the at least one fluorescent dye from the hydro-gel into the tissue region of interest.

9. The method of claim 8, wherein the light transparent matrix extends from the distal end of the fiber optic bundle by a distance of less than 100 μ

m.

10. The method of claim 8, wherein the light transparent matrix extends from the distal end of the fiber optic bundle by a distance of less than 200 μ

m.

11. The method of claim 1, wherein the at least one fluorescent dye is suspended in a buffer solution.

12. The method of claim 11, wherein the at least one fluorescent dye and the buffer solution comprises at least 95% of the dye carrier.

13. The method of claim 1, wherein each fluorescent dye of the at least one fluorescent dye is selected from a group consisting of:

Alexa, Texas Red, FITC, Oregon Green, Rhodamine Green, Lucifer yellow, Fluo 3, Fluo 4, and di-8-Anepps.

14. The method of claim 1, wherein each fluorescent dye of the at least one fluorescent dye has a molecular weight of 40 KDa or less.

15. The method of claim 14, wherein each fluorescent dye of the at least one fluorescent dye has a molecular weight of 20 KDa or less.

16. The method of claim 1, wherein each fluorescent dye of the at least one fluorescent dye has a molecular weight of between about 3 KDa and about 10 KDa.

17. The method of claim 1, wherein the tissue region of interest is cardiac tissue.

18. A catheter for developing a confocal image of a tissue region of interest of a subject in a confocal microscopic imaging system comprising a source of light energy that is configured to selectively generate light energy at a desired wavelength, the catheter comprising:

a fiber-optic bundle having a distal end and an opposed proximal end, the proximal end of the fiber-optic bundle being configured for operative communication with the source of light energy of the confocal microscopic imaging system; and

a dye carrier coupled to the distal end of the fiber-optic bundle, the dye carrier comprising at least one fluorescent dye, the dye carrier being coupled to the distal end of the fiber-optic bundle such that the dye carrier substantially covers the distal end of the fiber-optic bundle,

wherein, upon positioning of a portion of the dye carrier in contact with the tissue region of interest, the dye carrier is configured to permit diffusion of at least a portion of the at least one fluorescent dye into the tissue region of interest,

wherein the fiber-optic bundle is configured to transmit the light energy through the dye carrier and then into the tissue region of interest to excite the fluorescent dye within the tissue region of interest.

19. The catheter of claim 18, wherein the dye carrier comprises a light transparent matrix.

20. The catheter of claim 19, wherein the at least one fluorescent dye is suspended in a buffer solution.

21. The catheter of claim 20, wherein the at least one fluorescent dye and the buffer solution comprises at least 95% of the dye carrier.

22. The catheter of claim 19, wherein the at least one fluorescent dye is at a predetermined concentration.

23. The catheter of claim 19, wherein the dye carrier comprises at least one antibody agent.

24. The catheter of claim 19, further comprising a catheter sheath that is configured to selectively and at least partially enclose a distal end portion of the fiber optic bundle.

25. The catheter of claim 24, wherein the catheter sheath is configured to selectively and at least partially enclose at least a portion of the dye carrier.

26. The catheter of claim 18, wherein the at least one fluorescent dye is selected from a group consisting of:

Alexa, Texas Red, FITC, Oregon Green, Rhodamine Green, Lucifer yellow, Fluo 3, Fluo 4, and di-8-Anepps.

27. A catheter for developing a confocal image of a tissue region of interest of a subject in a confocal microscopic imaging system comprising a source of light energy that is configured to selectively generate light energy at a desired wavelength, the catheter comprising:

a fiber-optic bundle having a distal end and an opposed proximal end, the proximal end of the fiber-optic bundle being configured for operative communication with the source of light energy of the confocal microscopic imaging system; and

a dye carrier coupled to the distal end of the fiber-optic bundle, the dye carrier comprising at least one fluorescent dye and a light transparent matrix, the light transparent matrix comprising a hydro-gel,

wherein the hydro-gel is configured to permit diffusion of the at least one fluorescent dye from the hydro-gel into the tissue region of interest at a selected rate,

wherein the at least one fluorescent dye is configured to diffuse through the tissue region of interest, and

wherein the fiber-optic bundle is configured to transmit light into the tissue region of interest to excite the fluorescent dye within the tissue region of interest.

28. The catheter of claim 27, further comprising a catheter sheath that is configured to selectively and at least partially enclose a distal end portion of the fiber optic bundle.

29. The catheter of claim 28, wherein the catheter sheath is configured to selectively and at least partially enclose at least a portion of the dye carrier.

30. The catheter of claim 27, wherein the dye carrier is coupled to the distal end of the fiber-optic bundle such that the dye carrier substantially covers the distal end of the fiber-optic bundle, wherein the fiber-optic bundle is configured to transmit light through the dye carrier and then into the tissue region of interest to excite the fluorescent dye within the tissue region of interest.