Systems and methods for deep spectroscopic imaging of biological samples with use of an interferometer and spectrometer

• US 9,823,127 B2
• Filed: 04/09/2015
• Issued: 11/21/2017
• Est. Priority Date: 01/22/2010
• Status: Active Grant

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Wax, Adam, Levinson, Howard, Brown, William J., Matthews, Thomas, Medina, Manuel
• Primary Examiner(s): Lapage, Michael P
• Application Number:
  US14/683,066
• Publication Number:
  US 20150285685A1
• Time in Patent Office: 957 Days
• CPC Class Codes: A61B 5/445 Evaluating skin irritation or skin trauma, e.g. rash, eczema, wound, bed sore
• CPC Class Codes: G01B 9/02044 Imaging in the frequency domain, e.g. by using a spectrometer
• CPC Class Codes: G01B 9/02091 Tomographic interferometers, e.g. based on optical coherence
• CPC Class Codes: G01J 3/0218 using optical fibers
• CPC Class Codes: G01J 3/2823 Imaging spectrometer
• CPC Class Codes: G01J 3/453 by correlation of the amplitudes
• CPC Class Codes: G01J 3/4531 Devices without moving parts
• CPC Class Codes: G01N 21/45 using interferometric methods; using Schlieren methods
• CPC Class Codes: G01N 21/4795 spatially resolved investigating of object in scattering medium in vivo A61B

Abstract

Disclosed herein are systems and methods for deep spectroscopic imaging of a biological sample. In an aspect, a system includes a broad bandwidth light source configured to generate an illumination beam, an interferometer, and a spectrometer. The interferometer includes a first beam splitter configured to split the illumination beam into an incident beam and a reference beam; an optical lens directs the incident beam onto a biological sample at a predefined offset from corresponding optical axis, and receive a beam scattered from the biological sample. The beams are configured to intersect with each other within a focal zone of the optical lens. Photons of the incident beam undergo multiple forward scattering within the biological sample. A second beam splitter configured to receive and superimpose the scattered and reference beams, to generate an interference beam. The spectrometer uses a spectral domain detection technique to assess tissue properties of the biological sample.

Claims

1. A system for depth resolved spectroscopic imaging in deep tissue samples, the system comprising:

a broad bandwidth light source configured to generate an illumination beam;

an interferometer comprising;

a first beam splitter configured to receive and to split the illumination beam into an incident beam and a reference beam;

an optical element configured to;

receive the incident beam that is offset in distance from an optical axis resulting in a beam converging towards a focal zone situated at a predefined distance beneath the surface of a biological sample, wherein the converging beam is centered at an offset angle relative to the corresponding central optical axis; and

receive light scattered from the biological sample centered about an offset angle substantially equal to that of the incident beam and offset in the opposite direction;

a second beam splitter configured to;

receive and superimpose the scattered light and reference beam to generate an interference beam such that the spatial extent of the reference beam limits the received scattered light to that which has originated from the focal zone of the optical element; and

a spectrometer configured to receive the interference beam; and

a processor that uses a spectral domain detection technique to assess one or more depth resolved tissue properties of the biological sample based on a spectral interference pattern of the interference beam.

2. The system as claimed in claim 1, wherein the at least one broad bandwidth light source is one of a supercontinuum light source, a tungsten lamp, a superluminescent diode, and a swept source laser.

3. The system as claimed in claim 1, wherein the spectrometer includes a diffraction grating coupled to a charge coupled device (CCD) camera.

4. The system as claimed in claim 1, wherein the biological sample is burned skin tissue, and the deep spectroscopic imaging of the burned skin tissue generates information about tissue composition and structure.

5. The system as claimed in claim 1, wherein the incident and scattered beams are configured to have distinct illumination and detection apertures respectively.

6. The system as claimed in claim 1, wherein the interferometer is configured to employ coherence gating to restrict path length of one or more detected photons, and to employ spatial gating to localize paths of one or more detected photons, to enable the spectrometer to achieve imaging up to 90 Mean free paths (MFPs) and up to one centimeter depth of the biological sample.

7. The system as claimed in claim 6, wherein the incident and scattered beams are delivered and collected via one or more optical fibers.

8. The system as claimed in claim 1, wherein the data acquired from the spectrometer includes acquisitions of less than 10 milliseconds which are averaged together over a period of one or more seconds to one or more minutes to produce high dynamic range.

9. The system as claimed in claim 1, wherein the spectrometer is further configured to:

generate one or more one dimensional depth scans corresponding to one or more tissue layers of the biological sample, based on the spectral interference pattern of corresponding one or more interference beams; and

generate spectroscopic information of the biological sample based on a dual window method applied to the spectral interference pattern, wherein one or more windows are applied to the spectral interference pattern for creating corresponding one or more Short Time Fourier Transforms (STFTs), and combining the one or more STFTs to produce high resolution in pathlength and spectrum.

10. The system as claimed in claim 9, wherein the spectrometer is further configured to average the one or more one dimensional depth scans for generating the spectroscopic information with an improved Signal to Noise Ratio (SNR).

11. The system as claimed in claim 1, further comprising a modulator configured to modulate the incident beam at a predetermined modulation frequency.

12. The system of claim 11, wherein the modulator comprises one of a chopper wheel, a shutter, an electrooptic modulator, and an acousto optic modulator.

13. The system as claimed in claim 11, wherein the spectrometer is further configured to:

compute temporal characteristics for each depth point in a one dimensional depth scan, using Fast Fourier Transform (FFT) methods; and

isolate frequency bin in the FFT corresponding to the modulation frequency and rotate the isolated frequency bin by a predetermined modulation phase to suppress zero frequency artifact and autocorrelation features in the one dimensional depth scan.

14. The system as claimed in claim 1, wherein the interferometer further comprises an Optical Coherence Tomography (OCT) illumination source configured to generate an OCT beam positioned between the illumination and detection apertures.

15. The system of claim 1, further comprising a moving mirror that scans the converging beam and center offset angle of scattered light collection.

16. The system of claim 1, wherein the scattered light is collected centered around an offset angle that is equal magnitude but opposite direction relative to the optical axis as the converging beam.

17. The system as claimed in claim 1, wherein scattered light is collected at multiple directions, each with an offset angle that is equal magnitude as that of the incident converging beam but different directions relative to the optical axis.

18. The system as claimed in claim 17, further comprising one or more optical fibers that deliver and collect the incident and scattered beams are delivered and collected.

19. A method for depth resolved spectroscopic imaging in deep tissue samples, the method comprising:

generating an illumination beam;

splitting the illumination beam into an incident beam and a reference beam;

using an optical element to receive the incident beam that is offset in distance from the optical axis to produce a beam converging towards a focal zone situated at a predefined distance beneath the surface of a biological sample, where the converging beam is centered at an offset angle relative to the corresponding optical axis; and

receiving light scattered from the biological sample centered about an offset angle equal to that of the incident beam but offset in the opposite direction;

using a second beam splitter to receive and superimpose the scattered light and reference beam to generate an interference beam such that the spatial extent of the reference beam limits the received scattered light to that which has originated from the focal zone of the optical element;

superimposing the scattered and reference beams for generating an interference beam; and

employing a spectral domain detection technique for assessing one or more depth resolved tissue properties of the biological sample based on a spectral interference pattern of the interference beam.

20. The method as claimed in claim 19, wherein the incident and scattered beams are configured to have distinct illumination and detection apertures respectively.

21. The method as claimed in claim 19 further comprising:

generating one or more one dimensional depth scans corresponding to one or more tissue layers of the biological sample, based on the spectral interference pattern of corresponding one or more interference beams; and

generating spectroscopic information of the biological sample based on a dual window method applied to the spectral interference pattern, wherein one or more windows are applied to the spectral interference pattern for creating corresponding one or more Short Time Fourier Transforms (STFTs), and combining the one or more STFTs to produce high resolution in pathlength and spectrum.

22. The method as claimed in claim 21 further comprising averaging the one or more one dimensional depth scans for generating the spectroscopic information with an improved Signal to Noise Ratio (SNR).

23. The method as claimed in claim 22 further comprising:

computing temporal characteristics for each depth point in a one dimensional depth scan, using Fast Fourier Transform (FFT) methods; and

isolate frequency bin in the FFT corresponding to a modulation frequency and rotate the isolated frequency bin by a predetermined modulation phase to suppress the zero frequency artifact and autocorrelation features in the one dimensional depth scan.

24. The method as claimed in claim 19 further comprising employing coherence gating to restrict path length of the one or more photons, employing spatial gating to localize paths of the one or more photons, to achieve imaging up to 90 mean free paths (MFPs) and up to one centimeter depth of the biological sample.

25. The method as claimed in claim 19, further comprising using one or more optical fibers to deliver and collect the incident and scattered beams.

26. The method of claim 19 further comprising:

generating one or more one dimensional depth scans corresponding to one or more tissue layers of the biological sample based on the spectral interference pattern of corresponding one or more interference beams; and

generating spectroscopic information of the biological sample based on a dual window technique applied to the spectral interference pattern, wherein one or more windows are applied to the spectral interference pattern for creating corresponding one or more Short Time Fourier Transforms (STFTs), and combining the one or more STFTs to produce high resolution in pathlength and spectrum.

27. The method of claim 26 further comprising:

computing temporal characteristics for each depth point in a one dimensional depth scan, using Fast Fourier Transform (FFT) techniques; and

isolating frequency bin in the FFT corresponding to a modulation frequency and rotate the isolated frequency bin by a predetermined modulation phase to suppress the zero frequency artifact and autocorrelation features in the one dimensional depth scan.

28. The method of claim 19, further comprising using one or more optical fibers to deliver and collect the incident and scattered beams.

29. The method of claim 19, further comprising using a scanning mechanism to scan the converging beam and the central offset angle of scattered light collection such that the focal zone is translated across the sample and the offset angles of both are preserved.