Assessing blood brain barrier dynamics or identifying or measuring selected substances, including ethanol or toxins, in a subject by analyzing Raman spectrum signals

• US 7,398,119 B2
• Filed: 03/25/2003
• Issued: 07/08/2008
• Est. Priority Date: 07/13/1998
• Status: Expired due to Fees

• Current Assignee: California Institute of Technology, Childrens Hospital Los Angeles
• Original Assignee: California Institute of Technology, Childrens Hospital Los Angeles
• Inventors: Borchert, Mark S., Lambert, James L.
• Primary Examiner(s): Casler; Brian L.
• Assistant Examiner(s): MEHTA, PARIKHA SOLANKI
• Application Number:
  US10/396,035
• Publication Number:
  US 20040019283A1
• Time in Patent Office: 1,932 Days
• Field of Search: 600/318, 600/473, 356/301, 356/306
• US Class Current: 600/473
• CPC Class Codes: A61B 5/14532 for measuring glucose, e.g. by tissue impedance measurement
• 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 5/4064 Evaluating the brain A61B5/031, A61B5/369, A61B5/14553 take precedence
• CPC Class Codes: A61B 5/411 Detecting or monitoring allergy or intolerance reactions to an allergenic agent or substance
• CPC Class Codes: G01N 21/65 Raman scattering

Abstract

A non-invasive method for analyzing the blood-brain barrier includes obtaining a Raman spectrum of a selected portion of the eye and monitoring the Raman spectrum to ascertain a change to the dynamics of the blood brain barrier.

Also, non-invasive methods for determining the brain or blood level of an analyte of interest, such as glucose, drugs, alcohol, poisons, and the like, comprises: generating an excitation laser beam at a selected wavelength (e.g., at a wavelength of about 400 to 900 nanometers); focusing the excitation laser beam into the anterior chamber of an eye of the subject so that aqueous humor, vitreous humor, or one or more conjunctiva vessels in the eye is illuminated; detecting (preferably confocally detecting) a Raman spectrum from the illuminated portion of the eye; and then determining the blood level or brain level (intracranial or cerebral spinal fluid level) of an analyte of interest for the subject from the Raman spectrum. In certain embodiments, the detecting step may be followed by the step of subtracting a confounding fluorescence spectrum from the Raman spectrum to produce a difference spectrum; and determining the blood level and/or brain level of the analyte of interest for the subject from that difference spectrum, preferably using linear or nonlinear multivariate analysis such as partial least squares analysis. Apparatus for carrying out the foregoing methods are also disclosed.

Claims

1. A method of non-invasively detecting the presence or absence of an analyte in the cerebral spinal fluid or brain and/or monitoring the blood brain barrier of a subject comprising the steps of:

generating an excitation bean at a wavelength of between about 400 to 900 nanometers;

focusing the excitation bean of said generating step into the eye of the subject so that a selected region is illuminated;

obtaining a Raman spectrum corresponding to the illuminated region; and

detecting the presence or absence of an analyte in the brain or cerebral spinal fluid and monitoring the blood brain baffler dynamics based on said detecting step when the subject is exposed to gravity deficient environments.

2. A method according to claim 1, wherein said detecting step is carried out by illuminating the aqueous humor of the eye.

3. A method according to claim 1, wherein said detecting step is carried out by illuminating the vitreous humor of the eye.

4. A method according to claim 1, wherein said detecting step is carried out by obtaining a first in vivo Raman spectrum of the aqueous humor of the subject then obtaining a second in vivo Raman spectrum of the aqueous humor of the subject and comparing the first and second Raman spectrums to perform at least one of:

(a) detecting an alteration in the function of the blood brain baffler;

(b) identifying the presence or absence of a selected substance in the brain or cerebral spinal fluid; and

(c) determining a brain or cerebral spinal fluid concentration level of a selected substance.

5. A method according to claim 1 wherein the Raman spectrum is obtained without physically contacting the eye of the subject.

6. A method of non-invasively detecting the presence or absence of an analyte in the cerebral spinal fluid or brain and/or monitoring the blood brain baffler of a subject, comprising the steps of:

generating an excitation beam at a wavelength of between about 400 to 900 nanometers;

focusing the excitation beam of said generating step into the eye of the subject so that a selected region is illuminated;

obtaining a Raman spectrum corresponding to the illuminated region; and

detecting the presence or absence of an analyte in the brain or cerebral spinal fluid and/or monitoring the blood brain baffler dynamics based on said detecting step, wherein said detecting step comprises assessing whether the dynamics of the blood brain baffler allow at least one selected analyte, which would normally be inhibited from traveling through the blood brain barrier, to pass into the intracranial spinal fluid through the blood brain barrier.

7. A method according to claim 6, wherein said method further comprises the steps of:

administering a non-specific marker to the subject, the non-specific marker being selected for its normal reluctance to pass through the blood brain barrier;

comparing the Raman spectrum from said obtaining step to predetermined reference spectra corresponding to the non-specific marker; and

identifying the presence of the non-specific marker in the obtained Raman spectrum thereby indicating that the dynamics of the blood brain barrier have changed.

8. A method according to claim 7, further comprising the step of introducing a cytotoxic agent to the subject after said identifying step.

9. A method of non-invasively detecting the presence or absence of an analyte in the cerebral spinal fluid or brain and/or monitoring the blood brain barrier of a subject, comprising the steps of:

generating an excitation beam at a wavelength of between about 400 to 900 nanometers;

focusing the excitation beam of said generating step into the eye of the subject so that a selected region is illuminated;

obtaining a Raman spectrum corresponding to the illuminated region; and

detecting the presence or absence of an analyte in the brain or cerebral spinal fluid and monitoring the blood brain baffler dynamics based on said detecting step, wherein said detecting step is carried out when the subject is exposed to elevated ambient pressures.

10. A method of non-invasively detecting the presence or absence of an analyte in the cerebral spinal fluid or brain and/or monitoring the blood brain barrier of a subject, comprising the steps of:

generating an excitation beam at a wavelength of between about 400 to 900 nanometers;

focusing the excitation beam of said generating step into the eye of the subject so that a selected region is illuminated;

obtaining a Raman spectrum corresponding to the illuminated region; and

detecting the presence or absence of an analyte in the brain or cerebral spinal fluid and monitoring the blood brain baffler dynamics based on said detecting step, wherein said method is carried out when the brain of the subject has been exposed to a trauma injury to determine whether there is an alteration of the dynamics of the blood brain baffler.

11. A method according to claim 10, wherein said method is carried out during drug trials to assess the impact of the drug on the dynamics of the blood brain baffler of the subject.

12. A computer program product for monitoring the blood brain barrier and/or determining the presence and/or quantity of a selected substance in the brain of a subject, comprising:

a computer readable storage medium having computer readable program code embodied in said medium, said computer-readable program code comprising;

computer readable program code configured to analyze a Raman spectral response of a selected region of the eye of a subject to monitor the blood brain barrier and/or determine the presence and/or quantity of a selected substance in the intracranial and/or cerebral spinal fluid of a subject; and

computer program code configured to assess whether the dynamics of the blood brain barrier allow at least one selected analyte, which would normally be inhibited from traveling through the blood brain barrier, to pass into the intracranial spinal fluid through the blood brain barrier.

13. A computer program product according to claim 12, further comprising computer program code configured to compare the Raman spectral response to a priori data using a hybrid linear analysis technique.

14. A computer program product according to claim 12, further comprising computer program code configured to compare the Raman spectral response to a priori data using a partial least squares technique.

15. A computer program product according to claim 12, wherein the program code is configured to analyze a selected one or more peaks of interest in the Raman spectral response to identify the presence and/or quantity of the substance in the subject.

16. A computer program product according to claim 15, wherein the computer program product is configured to analyze a chemical shift peak associated with a carbon-carbon double bond on the molecular structure of the analyte of interest to identify tile concentration of the substance in the subject.