Method for the non-invasive measurement of tissue function and metabolism by determination of steady-state fluorescence anisotropy

• US 9,215,982 B2
• Filed: 11/09/2012
• Issued: 12/22/2015
• Est. Priority Date: 04/13/2006
• Status: Active Grant

• Current Assignee: Cellview Imaging Inc.
• Original Assignee: Cellview Imaging Inc.
• Inventors: Zuckerman, Ralph
• Primary Examiner(s): Singh, Satyendra K
• Application Number:
  US13/673,377
• Publication Number:
  US 20130072769A1
• Time in Patent Office: 1,138 Days
• US Class Current: 1/1
• CPC Class Codes: A61B 5/0071 by measuring fluorescence emission
• 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 5/14555 specially adapted for the eye fundus
• CPC Class Codes: A61B 5/14556 by fluorescence A61B5/14555 takes precedence
• CPC Class Codes: A61B 5/4842 Monitoring progression or stage of a disease
• CPC Class Codes: A61B 5/7246 using correlation, e.g. template matching or determination of similarity
• CPC Class Codes: A61B 5/7278 Artificial waveform generation or derivation, e.g. synthesising signals from measured signals A61B5/327 takes precedence
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 21/6445 Measuring fluorescence polarisation
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 2201/0683 Brewster plate; polarisation controlling elements
• CPC Class Codes: G01N 2201/12 Circuits of general importance; Signal processing

Abstract

A non-invasive measurement of biological tissue reveals information about the function of that tissue. Polarized light is directed onto the tissue, stimulating the emission of fluorescence, due to one or more endogenous fluorophors in the tissue. Fluorescence anisotropy is then calculated. Such measurements of fluorescence anisotropy are then used to assess the functional status of the tissue, and to identify the existence and severity of disease states. Such assessment can be made by comparing a fluorescence anisotropy profile with a known profile of a control.

Claims

1. A method for non-destructively evaluating the functional capacity of a human tissue using a signal processing apparatus configured for non-invasive evaluation of the tissue in a clinical setting, the method comprising:

irradiating the tissue in a resting state with polarized light so as to cause one or more endogenous fluorophores in the tissue to fluoresce, the one or more endogenous fluorophores comprising lipoamide dehydrogenase (LipDH);

determining a resting steady-state fluorescence anisotropy of emitted fluorescence in tissue in the resting state;

stimulating the tissue to change its metabolism from the resting state into a stimulated state;

irradiating the tissue in the stimulated stated with polarized light so as to cause one or more endogenous fluorophores in the tissue to fluoresce;

determining a stimulated steady-state fluorescence anisotropy of emitted fluorescence in the tissue in the stimulated state;

constructing a resting fluorescence anisotropy map based on the resting steady-state fluorescence anisotropy and a stimulated fluorescence anisotropy map based on the stimulated steady-state fluorescence anisotropy;

determining a the functional capacity of the tissue by subtracting point by point or pixel by pixel the resting fluorescence anisotropy map from the stimulated fluorescence anisotropy map over the entire maps or within an area of interest; and

comparing the functional capacity of the tissue to a functional capacity of a control tissue similarly evaluated to identify a tissue dysfunction.

2. The method of claim 1, wherein the step of stimulating the tissue to change its metabolism comprises administering oxygen to said tissue.

3. The method of claim 1, further comprising:

measuring steady-state fluorescence anisotropy values for a plurality of locations at a plurality of depths in the tissue in resting and stimulated states;

constructing a resting three-dimensional map of steady-state fluorescence anisotropy based on the steady-state fluorescence anisotropy values for the tissue in the resting state and a stimulated three-dimensional map of steady-state fluorescence anisotropy based on the steady-state fluorescence anisotropy values for the tissue in the stimulated state; and

determining the functional capacity of the tissue by subtracting point by point or pixel by pixel the resting three-dimensional map from the stimulated three-dimensional map over the entire maps or within an area of interest.

4. The method of claim 1, wherein the tissue dysfunction comprises a presence and severity of a disease.

5. The method of claim 4, wherein the disease comprises cancer.

6. The method of claim 4, wherein the disease is selected from the group consisting of ocular posterior pathologies including diabetic retinopathy, age-related macular degeneration and primary open angle glaucoma, and other optic neuropathies.

7. The method of claim 1, wherein the stimulating comprises irradiating the tissue with light and the tissue is selected from the group consisting of skin tissue, cervix tissue, abdominal tissue, esophagus tissue, stomach tissue, gut tissue, lung tissue, and ocular tissue.

8. The method of claim 1, further comprising determining, based at least in part on the functional capacity of the tissue, an efficacy of a treatment intervention.

9. The method of claim 1, wherein the step of stimulating the tissue to change its metabolism comprises irradiating the tissue with steady-state light.

10. The method of claim 9, wherein a wavelength of the steady-state light is selected from a range between 400 nm and 700 nm.

11. The method of claim 1, wherein stimulating the tissue to change its metabolism is performed using stimulation having at least one different property than properties of the polarized light used for irradiating the tissue.