Apparatus and method for fluorescent excitation and detection from potentiometric dyes with a single-ended optical fiber

• US 5,239,998 A
• Filed: 07/18/1991
• Issued: 08/31/1993
• Est. Priority Date: 07/18/1991
• Status: Expired due to Term

• Current Assignee: The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Inventors: Krauthamer, Victor
• Primary Examiner(s): Cohen, Lee S.
• Assistant Examiner(s): PONTIUS, KEVIN LANE
• Application Number:
  US07/732,021
• Time in Patent Office: 775 Days
• Field of Search: 128/633-634, 128/664-665, 356/36, 356/402
• US Class Current: 600/317
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence

Abstract

A method of detecting and recording action-potential-related fluorescent changes from a remote tissue situs which has been treated with a voltage-sensitivity dye. The method utilizes a single optical fiber to both direct excitation light to the remote tissue situs and receive emitted fluorescence from the remote tissue situs. The emitted fluorescence from the remote tissue situs is directed to a photomultiplier or photodiode which produces a resulting fluorescence-related signal which is analyzed. The method and related apparatus eliminates signal noise and allows accurate recording and analysis of action-potential-related fluorescent changes.

Claims

1. A method of detecting electrical characteristics of a remote sample situs which comprises:

treating the remote sample situs with a voltage-sensitive dye providing a single optical fiber and a detector;

delivering light of a sufficient wavelength to cause fluorescent emission of said dye-treated remote sample situs to said dye-treated remote sample situs by means of said single optical fiber; and

receiving fluorescent emission from said dye-treated remote sample situs by said single optical fiber and delivering said received fluorescent emission to said detector for analyzing said received fluorescent emission.

2. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said remote sample situs comprises a tissue sample.

3. A method of detecting electrical characteristics of a remote sample situs according to claim 2, wherein said tissue sample is within a subject'"'"'s body.

4. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said detected electrical characteristics comprise electrical activity of said remote sample situs.

5. A method of detecting electrical characteristics of a remote sample situs according to claim 4, wherein said electrical characteristics comprise action-potential-related changes in said remote sample situs.

6. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said single optical fiber has a core diameter of between about 50 to 100 μ

m.

7. A method of detecting electrical characteristics of a remote sample situs according to claim 6, wherein said single optical fiber includes a cladding layer and an overall diameter of up to about 140 μ

m.

8. A method of detecting electrical characteristics of a remote sample situs according to claim 1, further comprising directing said light of a sufficient wavelength to cause fluorescent emission of said dye-treated remote sample situs though a neutral density filter to attenuate said light.

9. A method of detecting electrical characteristics of a remote sample situs according to claim 8, further comprising directing said light of a sufficient wavelength to cause fluorescent emission of said dye-treated remote sample though a short-pass interference filter to remove light of an interfering wavelength.

10. A method of detecting electrical characteristics of a remote sample situs according to claim 9, further comprising directing said received fluorescent emission to said detector utilizing a beam splitter.

11. A method of detecting electrical characteristics of a remote sample situs according to claim 10, further, comprising directing said received fluorescent emission through a long-pass filter to remove any excitation light from said received fluorescent emission.

12. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said dye treated remote sample situs is treated with a voltage-sensitive dye.

13. A method of detecting electrical characteristics of a remote sample situs according to claim 12, wherein said voltage-sensitive dye comprises a styryl voltage-sensitive dye.

14. A method of detecting electrical characteristics of a remote sample situs according to claim 13, wherein the wavelength of said light delivered to said dye-treated remote sample is between about 490 to 550 μ

m.

15. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said detector comprises a photomultiplier.

16. A method of detecting electrical characteristics of a remote sample situs according to claim 1, wherein said detector comprises a photodiode.

17. An apparatus for detecting electrical characteristics of a remote sample situs which comprises:

a light source for generating excitation light having a wavelength of between about 400 to 550 nm;

means to pass said excitation light through a single optical fiber to a remote sample situs;

means interposed between said light source and said single optical fiber for attenuating said excitation light;

means interposed between said attenuating means and said single optical fiber for removing light of an interfering wavelength from said excitation light;

a fluorescent detector;

means for directing emitted light from said remote sample situs through said single optical fiber to said fluorescent detector; and

means positioned between said directing means and said fluorescent detector for removing light of an excitation wavelength from said emitted light.

18. An apparatus for detecting electrical characteristics of a remote sample situs according to claim 17, wherein said single optical fiber has a core diameter of between about 50 to 100 μ

m.

19. An apparatus for detecting electrical characteristics of a remote sample situs according to claim 18, wherein said single optical fiber includes a cladding layer and an overall diameter of up to about 140 μ

m.

20. An apparatus for detecting electrical characteristics of a remote sample situs according to claim 17, wherein said fluorescent detector comprises a photomultiplier.