Systems and methods for imaging fluorophores

• US 6,304,771 B1
• Filed: 01/15/1997
• Issued: 10/16/2001
• Est. Priority Date: 10/29/1993
• Status: Expired due to Term

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Chance, Britton, Li, Xingde, Boas, David A., OʼLeary, Maureen, Yodh, Arjun G.
• Primary Examiner(s): Casler, Brian L.
• Application Number:
  US08/783,682
• Time in Patent Office: 1,735 Days
• Field of Search: 600/476, 600/477, 600/309, 600/310, 356/39, 356/319, 356/346, 356/432, 356/436, 356/441, 356/442, 356/446, 356/239, 356/318, 250/358.1, 250/574, 250/575, 250/458.1, 250/459.1
• US Class Current: 600/476
• 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: G01N 21/4795 spatially resolved investigating of object in scattering medium in vivo A61B
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging

Abstract

The systems comprise source means for illuminating the fluorophores with diffuse photon density waves of a first specified wavelength, whereby the fluorophores will fluoresce re-radiated diffuse photon density waves of a second wavelength after being illuminated with the diffuse photon density waves of the first specified wavelength; detection means for detecting the re-radiated diffuse photon density waves of the second wavelength, wherein there is a phase shift between the diffuse photon density waves of the first wavelength and the diffuse photon density waves of the second wavelength; and processing means interfaced with the detection means for processing data corresponding to the phase shift and the amplitude of the re-radiated waves to determine concentration and lifetime of the fluorophores as a function of spatial position of the fluorophores in the turbid medium.

Claims

1. A system for imaging fluorophores in a turbid medium comprising:

an illumination source that illuminates the fluorophores with diffuse photon density waves of a first specified wavelength, thus causing the fluorophores to fluoresce re-radiated diffuse photon density waves of a second wavelength after being illuminated with the diffuse photon density waves of the first specified wavelength;

a detector that detects the re-radiated diffuse photon density waves of the second wavelength, wherein there is a phase shift between the diffuse photon density waves of the first wavelength and the diffuse photon density waves of the second wavelength; and

a processor interfaced with the detector that processes data corresponding to the phase shift and the amplitude of the re-radiated waves to determine concentration and lifetime of the fluorophores as a function of spatial position in the turbid medium.

2. The system recited in claim 1 wherein the illumination source comprises at least one laser oriented around the fluorophores which irradiates the fluorophores with the diffuse photon density waves of the first wavelength to cause the fluorophores to fluoresce at the second wavelength.

3. The system recited in claim 2 wherein the detector comprises an optical fiber that is placed in proximity to the fluorophores and a photomultiplier tube interfaced to the optical fiber.

4. The system recited in claim 3 further comprising a switch interfaced with the laser that alternately and sequentially turns on and off the laser, and a radio frequency driver interfaced through the switch with the laser that drives the laser to produce the diffuse photon density waves of the first wavelength.

5. The system recited in claim 4 further comprising a display interfaced with the processor that displays the image of the fluorophores'"'"' concentration and lifetime produced by the processor.

6. The system recited in claim 1 wherein the illumination source comprises at least one light emitting diode oriented around the fluorophores which irradiates the fluorophores with the diffuse photon density waves of the first wavelength to cause the fluorophores to fluoresce at the second wavelength.

7. The system recited in claim 1 comprising two illumination sources located symmetrically about the detector.

8. The system recited in claim 1 comprising two detectors located symmetrically about the illumination source.

9. The system recited in claim 1 further comprising means for initially providing information about the position and lifetime of the fluorophores in the turbid medium and wherein the processor is adapted to process the data to further characterize the fluorophores in the turbid medium, wherein the process applies an analytic forward solution.

10. The system recited in claim 1 wherein the processor processes the data to determine at least the position and lifetime of the fluorophore in the turbid medium, wherein the processor further applies a perturbative inverse processing procedure.

11. A method imaging a fluorophores in a turbid medium comprising the steps of:

illuminating the fluorophores with diffuse photon density waves of a first specified wavelength;

allowing the fluorophores to fluoresce, thereby re-radiating a diffuse photon density wave having a second wavelength and an amplitude;

detecting the re-radiated diffuse photon density wave having the second wavelength and amplitude wherein there is a phase shift between the diffuse photon density wave of the first wavelength and the diffuse photon density wave of the second wavelength; and

analyzing the phase difference and the amplitude to determine concentration and lifetime of the fluorophores as a function of spatial position in the turbid medium.

12. The method recited in claim 11 wherein the illuminating step comprises the step of alternately irradiating the fluorophores with at least two sources of diffuse photon density waves of the first wavelength to cause the fluorophores to fluoresce.

13. The method recited in claim 11 wherein the illuminating step comprises the step of illuminating the fluorophores with at least one laser.

14. The method recited in claim 11 wherein the illuminating step comprises the step of illuminating the fluorophores with at least one light-emitting diode.

15. The method recited in claim 11 further comprising the step of initially providing information about the position of the fluorophores in the turbid medium, and the analyzing step further characterizes the fluorophores in the turbid medium.

16. The method recited in claim 11 wherein the analyzing step determines at least the position of the fluorophores in the turbid medium.