WO2007082228A1

WO2007082228A1 - Systems and methods for generating data based on one or more spectrally-encoded endoscopy techniques

Info

Publication number: WO2007082228A1
Application number: PCT/US2007/060319
Authority: WO
Inventors: Guillermo J. Tearney; Seok-Hyun Yun; Brett Eugene Bouma; Dvir Yelin
Original assignee: The General Hospital Corporation
Priority date: 2006-01-10
Filing date: 2007-01-10
Publication date: 2007-07-19
Also published as: JP2009523040A; US20070263208A1; EP1971848A1; US7796270B2; EP1971848B1; JP5680826B2

Abstract

Exemplary systems and methods for generating data associated with at least one portion of a sample can be provided. For example, according to one exemplary embodiment of such systems and methods, it is possible to provide a particular radiation using at least one first arrangement. The particular radiation can include at least one first electro-magnetic radiation directed to at least one sample and at least one second electro-magnetic radiation directed to a reference arrangement. The first radiation and/or the second radiation can comprise a plurality of wavelengths. The first electro-magnetic radiation can be spectrally dispersed along at least one portion of the sample. The second electro-magnetic radiation measured at two or more different lengths of the reference arrangement with respect to the first arrangement. Data can be generated which is associated with the first and second electro-magnetic radiations obtained at the two different lengths using at least one second arrangement which comprises a spectrometer arrangement.

Description

SYSTEMS AND METHODS FOR GENERATING DATA BASED ON ONE OR MORE SPECTRALLY-ENCODED ENDOSCOPY TECHNIQUES

CROSS-REFERENCE TO RELATED APPLICATION(S)

This application is based upon and claims the benefit of priority from U.S. Patent Application Serial No. 60/757,569, filed January 10, 2006, the entire disclosure of which is incorporated herein by reference.

STATEMENT REGARDINGFEDERALLY SPONSORED RESEARCH

The invention was made with the U.S. Government support under Contract

No. BES-0086709 awarded by the National Science Foundation. Thus, the U.S. Government has certain rights in the invention.

FIELD OF THE INVENTION

The present invention generally relates to systems and methods for generating data associated with at least one portion of a sample, and particularly for, e.g., generating such data using one or more spectrally-encoded endoscopy techniques.

BACKGROUND OF THE INVENTION

As is known in the art, three-dimensional (3D) endoscopy which provides clinicians with depth information can greatly aid a variety of minimally invasive procedures. Depth resolved imaging with a large, three-dimensional field of view is, however, challenging when utilizing flexible imaging probes such as borescopes, laparoscopes, and endoscopes having relatively small diameters. Confocal imaging through a fiber-bundle using a high numerical aperture (NA) lens such as that described in Y. S. Sabharwal et al., "Slit-scanning confocal microendoscope for high-resolution in vivo imaging," Appl. Opt. 38, 7133 (1999) is one solution to this problem. The 3D field of view for these devices, however, may be limited to less than a few millimeters due to the small clear aperture of the objective lens and low f-number required for high-resolution optical sectioning.

Other methods, such as stereo imaging and structured illumination as described in M. Chan et al., "Miniaturized three-dimensional endoscopic imaging system based on active stereovision," Appl. Opt. 42, 1888 (2003) and D. Karadaglic et al., "Confocal endoscope using structured illumination," Photonics West 2003, Biomedical Optics, 4964-34, respectively, have also been described. These techniques, however, likely a large number of pieces of hardware for the probe than confocal imaging through a fiber bundle. This additional hardware increased the size, cost, and complexity of such devices.

Spectrally-encoded endoscopy ("SEE") is a techniques which uses a broadband light source and a diffraction grating to spectrally encode reflectance across a transverse line within a sample. A two-dimensional image of the sample can be formed by slowly scanning this spectrally-encoded line across the sample. This exemplary technique generally uses a single optical fiber, thereby enabling imaging through a flexible probe having a small diameter. When combined with interferometry, the SEE technique has the additional capability of providing three- dimensional images.

Depth-resolved imaging can be achieved by incorporating an SEE probe into the sample arm of a Michelson interferometer. Using this arrangement, two- dimensional ("2D") speckle patterns can be recorded by a charge-coupled device ("CCD") camera at multiple longitudinal locations of a reference mirror. Subsequently, depth information can be extracted by comparing the interference obtained at consecutive reference mirror positions.

One problem with this exemplary approach, however, may be that the reference mirror should be maintained in a stationary configuration to within an optical wavelength during a single image (or line) acquisition time to avoid the loss of fringe visibility. Stepping the reference mirror with such high fidelity over multiple discrete depths is quite challenging at the high rates required for real-time volumetric imaging.

One exemplary improvement in an acquisition speed and therefore susceptibility to sample motion can be achieved using time-domain heterodyne interferometry. In this exemplary approach, an interference signal can be recorded with a standard photodetector at every group delay scan of a rapid scanning optical delay line ("RSOD"). By applying a short-time Fourier transformation ("STFT") to a so-measured trace, transverse and depth information can be extracted. In this exemplary method, three-dimensional ("3D") data sets can be acquired at a rate of about five per second.

Image quality can be governed, at least in part, by the signal-to-noise ratio ("SNR"). For fast imaging rates and low illumination powers used for safe clinical imaging, maintaining a high SNR may be challenging. For example, with 4 milli-watt

(mW) of power on the sample, the time-domain SEE system can provide an SNR of approximately 10 dB. Using an exemplary SEE technique, a low NA collection system can be utilized to achieve a large working distance, likely resulting in a decrease in the solid angle of collection of light scattered from tissue. Typical optical parameter of NA=0.01 can allow the collection of, e.g., only 0.01% of light scattered from the sample. As a result, the signal of SEE images of tissue may, e.g., only be 10 dB above the noise floor. Increasing the imaging speed increases the bandwidth, thereby decreasing the SNR commensurately.

High imaging speed and high SNR can be important for clinical applications of optical imaging. Optical coherence tomography ("OCT") is an imaging technique which shares certain principles with 3D SEE. For example, exemplary OCT systems can utilize the RSOD and a single detector, and may be operated with an A-line acquisition rate in the range of about 2-3 kHz. This can correspond to about 4 frames/second with 500 A-lines per frame. An improvement of a number of orders of magnitudes in imaging speed has been demonstrated with an alternative approach, e.g., a spectral-domain OCT ("SD-OCT") technique, hi this exemplary technique, a high-resolution spectrometer can include a diffraction grating and a linear CCD array which may be used to record spectral interference between light from a sample and from a fixed-length reference arm. The improvement in SNR using such exemplary technique has enabled imaging at 30,000 A-lines per second, nearly three orders of magnitude improvement over time-domain methods.

An exemplary proof of principle using spectral domain interferometry for spectral encoding has been described in by Froehly et al., Optics Communications 222 (2003), pp. 127-136. In such document, transverse and depth resolution of this exemplary technique has been analyzed, and a phase-sensitive depth measurement of a 1 mm thick glass plate was demonstrated.

Accordingly, it may be beneficial to address and/or overcome at least some of the deficiencies described herein above.

OBJECTS AND SUMMARY OF THE INVENTION

One of the objectives of the present invention is to overcome certain deficiencies and shortcomings of the prior art arrangements and methods (including those described herein above), and provide exemplary embodiments of systems and methods for generating data associated with at least one portion of a sample, and particularly for, e.g., generating such data using one or more spectrally-encoded endoscopy techniques.

According to one exemplary embodiment of the systems and methods of the present invention, it is possible to provide a particular radiation using at least one first arrangement. The particular radiation can include at least one first electro-magnetic radiation directed to at least one sample and at least one second electro-magnetic radiation directed to a reference arrangement. The first radiation and/or the second radiation can comprise a plurality of wavelengths. The first electro-magnetic radiation can be spectrally dispersed along at least one portion of the sample. The second electro-magnetic radiation measured at two or more different lengths of the reference arrangement with respect to the first arrangement. Data can be generated which is associated with the first and second electro-magnetic radiations obtained at the two different lengths using at least one second arrangement which comprises a spectrometer arrangement. For example, the reference arrangement can include a translatable mirror arrangement, a piezo-electric fiber stretching arrangement, a pulse-shaping arrangement, a rapidly-scanning optical delay line arrangement and/or an electro- optical or acousto-optical arrangement. The data can be generated based on a Fourier transform of information received by the spectrometer arrangement which is associated with the first and second electro-magnetic radiations. According to one exemplary variant, the data may be generated as a function of a phase of the information.

According to another exemplary embodiment of the present invention, the data can be generated based on a time frequency transform and/or a space-frequency transform of information received by the spectrometer arrangement which is associated with the first and second electro-magnetic radiations. The transform can be a Short-time Fourier transform. The data may also be generated based on a correlation between information received at at least one first length of the two different lengths and at least second length of the two different lengths. The correlation can be a cross-correlation. A first peak of the cross-correlation may be obtained, and a sign of the cross-correlation can be determined based on further information associated with the first peak.

According to a further exemplary embodiment of the present invention, the data may be generated based on a comparison of phases between information received at at least one first length of the two different lengths and at least second length of the two different lengths. Further data can be generated based on a magnitude of a Fourier transform of the data. The data and the further data can be combined to form a composite image data associated with the sample. In another exemplary embodiment of the present invention, the data may be associated with at least one portion of the sample which is located in a direction that is axial with respect to a direction of the first electro-magnetic radiation. The sign of a location of the first peak of the cross-correlation may be associated with the portion of the sample which is located in the direction that is axial with respect to the direction of the first electro-magnetic radiation.

According to still another exemplary embodiment of the present invention, the data can be further associated with a two-dimensional image and/or a three- dimensional image of the portion of the sample. The second electro-magnetic radiation can be directed to continuously scan the different lengths of the reference arrangement.

In accordance with yet another exemplary embodiment of the present invention, a spectral-domain spectrally-encoded endoscopy ("SD-SEE") system and method can be provided. For example, a source and an interferometer can be utilized. The interferometer can have a first port coupled to receive a signal from the source, a second port coupled to reference arm which may include a path length control device, a third port coupled to a sample arm which may include a scanning element and one or more optical imaging elements, and a fourth port coupled to a detector which may include a Spectrometer and a camera.

The exemplary embodiment of the SD-SEE system can perform 3D imaging, e.g., at about 30 frames per second, with SNR of greater than about 30 dB. Using an exemplary high-speed line camera in a high-resolution spectrometer, e.g., each x-z plane can be captured in a single shot. This exemplary embodiment of the present invention facilitates high-speed imaging with two to three orders of magnitude improvement in SNR over conventional techniques. In one further exemplary embodiment of the present invention, the path length control device of the reference arm can include an RSOD, and real-time 3D imaging of tissue can be facilitated at a rate of about 30 frames per second with an SNR typically of about 30 dB. SNR of greater than about 30 dB can provide an improvement of about two to three orders of magnitude over time-domain SEE techniques.

In a further exemplary embodiment of the present invention, a detection SEE technique can be provided which uses spectral-domain interferometry. For example, a light or other electro-magnetic radiation can be provided to a sample and a reference arm. A path length of the reference arm can be changed, spectral interference between light reflected from the sample and light reflected from the reference arm may be measured, and a cross-correlation between a short time Fourier transforms (STFTs) of adjacent sample locations can be determined.

Using this exemplary arrangement according to one exemplary embodiment of the present invention, it is possible to resolve a depth ambiguity in SD-SEE. By determining a cross-correlation between STFTs of adjacent sample locations, the sign of an offset of the first cross-correlation maximum can be used to remove height ambiguity. The implementation of this exemplary cross-correlation technique can be accomplished by, e.g., stepping the sample arm slow scan axis to one location, acquiring one spectral scan, stepping the reference arm path length, and acquiring an additional spectral scan. This exemplary procedure may be repeated for the entire volumetric image. For example, moving either the sample arm or the reference path in small steps may be challenging at high speeds. Thus, according certain exemplary embodiments, the reference arm can be continuously scanned at a slow rate. The sample arm can also be continuously scanned and spectral data can be over-sampled by capturing a plurality of spectrally-encoded lines per resolution element, which can be determined by the numerical aperture of the SEE probe lens.

In accordance with a further exemplary embodiment of the present invention, the exemplary SD-SEE technique can include (a) stepping a sample arm slow scan axis to a location, (b) acquiring one spectral scan at the location, (c) stepping a reference arm path length, and (d) acquiring an additional spectral scan. If the entire volumetric image is complete, the processing can be stopped. Otherwise, the sample arm slow scan axis may be moved to a new location and processing steps (b) - (d) can be repeated.

Using an exemplary embodiment of the arrangement which uses such exemplary technique for spectral-domain spectrally-encoded endoscopy, a desirable SNR can be provided. For example, moving either the sample arm or the reference path (e.g., an RSOD galvanometer) in small steps may be challenging at high speeds. Thus, according to certain exemplary embodiments, the reference arm can be continuously scanned so that the optical delay between two adjacent lines in the image may be in the range of about 0 to λ/2, and preferably about λ/4.

In one additional exemplary embodiment of the present invention, the reference arm can be continuously scanned at a rate of about 30 Hz, with a sawtooth waveform having an amplitude typically of about 100 μm, the sample arm may be continuously scanned, and spectral data can be over-sampled by capturing about a plurality of spectrally-encoded lines per resolution element, which can be determined by the numerical aperture of the SEE probe lens. Another exemplary way to remove depth ambiguity may be by (a) acquiring a full 3D image at a fixed location of the reference arm, (b) moving the reference arm to another location, (preferably by one axial resolution element), and (c) acquiring second 3D image. The unwrap mask can be obtained by determining the sign of the difference between the two height maps (e.g., depth information only).

Other features and advantages of the present invention will become apparent upon reading the following detailed description of embodiments of the invention, when taken in conjunction with the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS Further objects, features and advantages of the present invention will become apparent from the following detailed description taken in conjunction with the accompanying figures showing illustrative embodiments of the present invention, in which:

FIG. 1. is a block diagram of an exemplary embodiment of a spectral-domain spectrally-encoded endoscope ("SD-SEE") imaging system having a rapidly scanning optical delay line ("RSOD") with neutral density ('TSTD");

FIG. 2. is a block diagram illustrating an exemplary embodiment of data capturing and processing in the SD-SEE system of FIG. 1;

FIG. 3A is an exemplary unprocessed image of a doll's face from which information may be extracted, and includes an inset corresponding to a white-light image of the doll's face; FIG. 3 B is an expanded view of a portion of the image of FIG. 3 A, as circumscribed by a white rectangle;

FIG. 3 C is an exemplary graph of image intensity versus distance taken along the solid and the dashed lines at a particular location X indicated in FIG. 3B;

FIG. 3D is an exemplary graph of image intensity versus distance taken along the solid and the dashed lines at another location Y indicated in FIG. 3B;

FIG. 3E is an exemplary graph of image intensity versus distance taken along the solid and the dashed lines at a further location Z indicated in FIG. 3B;

FIG. 3F is an exemplary graph of power spectra (power vs. frequency) taken along the solid and the dashed lines at a certain location X indicated in FIG. 3B;

FIG. 3G is an exemplary graph of power spectra (power vs. frequency) taken along the solid and the dashed lines at an additional location Y indicated in FIG. 3B;

FIG. 3H is an exemplary graph of power spectra (power vs. frequency) taken along the solid and the dashed lines at a further location Z indicated in FIG. 3B;

FIG. 31 is an exemplary graph of cross-correlation between the power spectra of adjacent (solid and dotted) lines at the location X in FIG. 3B;

FIG. 3 J is an exemplary graph of the corresponding cross-correlation between the power spectra of adjacent (solid and dotted) lines at the location Y in FIG. 3B;

FIG. 3K is an exemplary graph of the corresponding cross-correlation between the power spectra of adjacent (solid and dotted) lines at the location Z in FIG. 3B; Fig. 4A is an exemplary two-dimensional ("2D") image of the face of the doll;

FIG. 4B is an exemplary height map of the face of the doll shown in FIG. 4A, with numbers on the scale bar being millimeters;

FIG. 4C is an exemplary image of a sign mask of the face of the doll shown in FIG. 4A for unwrapping a depth image;

FIG. 4D is an exemplary unwrapped depth map of the face of the doll shown in FIG. 4A.

FIG. 5 A is are exemplary three-dimensional ("3D") images of a paper flower;

FIG. 5B are exemplary 3D images of a skin fold of a volunteer;

FIG. 5 C are exemplary 3D images of two hind paws and a tail of a mouse embryo, with the transverse 2D images, depth images, and depth being superimposed on the 2D image being shown on the left, middle and right images, respectively; and

FIG. 6 is a flow diagram describing an exemplary procedure for an extraction of three dimensional information in according to one exemplary embodiment of the present invention.

Throughout the figures, the same reference numerals and characters, unless otherwise stated, are used to denote like features, elements, components or portions of the illustrated embodiments. Moreover, while the subject invention will now be described in detail with reference to the figures, it is done so in connection with the illustrative embodiments. It is intended that changes and modifications can be made to the described embodiments without departing from the true scope and spirit oi me subject invention as defined by the appended claims.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS

FIG. 1 shows a block diagram of an exemplary embodiment of a system according to the present invention for spectral-domain spectrally-encoded endoscopy

("SD-SEE"). This exemplary system includes a light source 10 coupled to a first port

12a of a single-mode fiber optic interferometer 12. In one exemplary embodiment, the light source 10 may be, for example, a broad-bandwidth titanium-sapphire laser of the type manufactured by, e.g., Femtolasers Produktions, GmbH, Femtosource integral OCT™, with a center wavelength of about 800 nanometers (nm) and an

FWKfM bandwidth of about 140 nm. Further, according to another exemplary embodiment, the interferometer 12 may be a 50/50 Michelson interferometer.

A second port 12b of the interferometer 12 of the system of FIG. 1 can be coupled to a reference arm which may include, e.g., an arrangement for adjusting or otherwise controlling a path length 14 coupled thereto. A third port 12c of the interferometer 12 can be coupled to a sample arm having, e.g., a miniature endoscopic imaging probe 16 coupled thereto. A fourth port 12d of the interferometer 12 can be coupled to a detection arm having, e.g., a detector 18 coupled thereto.

In the exemplary embodiment of the system of FIG. 1, the arrangement which is capable of adjusting the path length 14 can include a rapidly scanning optical delay line ("RSOD") which may control a group delay of light or other electro-magnetic radiation propagating in the reference arm. The RSOD can be provided from a neutral density ("ND") filter, a grating (or other light/electro-magnetic radiation providing device), a lens and a galvanometric scanner.

The miniature endoscopic imaging probe 16 of FIG. 1 can be simulated utilizing a compact lens-grating design in which a beam can be first focused by a lens (e.g., f = 40 mm, beam diameter 0.5 mm), and then diffracted by a transmission grating (e.g., Holographix LLC, 1000 lines/mm) to form a spectrally-encoded line (x- axis) on the surface of a sample 20. The galvanometric optical scanner can be used to provide a slow (y) axis scanning. The above exemplary parameters can result in a spatial transverse resolution of approximately 80 μm. The image can be comprised of, e.g., about 80 transverse resolvable points; each transverse spot may be illuminated with a bandwidth of 1.9 nm. The overall power on the sample 20 can be about 4 mW.

The detector 16 may be a spectrometer. In one exemplary embodiment, a spectrometer can have a relatively high resolution, which may be comprised of a beam collimator (e.g., Oz optics, f=50 mm), a diffraction grating (e.g., Spectra- Physics, 1800 lines/mm), a lens (e.g., Nikon 85 mm, f/1.8) and a high-speed (e.g., 2048 elements, read-out rate 30 kHz) line scan camera (Basler L104k-2k). A function generator can be used to control the galvanometric scanner(s) at the sample arm and in the RSOD, and may provide a synchronization signal to the line scan camera.

FIG. 2 shows various exemplary images and graphs to illustrate and describe an exemplary embodiment of an image formation process. For example, a sample 50 to be imaged may have three surfaces 50a, 50b, 50c, with each of the three surfaces 50a, 50b, 50c having different heights, as shown in FIG. 2. An imaging probe (not shown in FIG. 2) with which the sample 50 can be imaged may provide three resolvable points at wavelengths λj, λ₂, λ₃. To promote clarity and for ease of explanation, it can be assumed that the sample 50 may have a uniform or substantially uniform reflectivity. Further, in certain practical imaging systems, the imaging probes utilizing more than three resolvable points may typically be used.

Each horizontal line recorded by the CCD (e.g. the CCD in the detector 18 of

FIG. 1) can correspond to a spectral interference between the light reflected from the sample 20 and from the reference arm. A series of interference patterns 52a-52c (plotted as a solid lines in FIG. 2) can have modulation frequencies which may be proportional to the absolute distance from a plane that can match the path length of the reference arm (e.g., can be referred to as a "zero plane").

As shown in FIG. 2, for example, the reference arm path length can match the distance from a surface (or plane) 50b. Thus, the interference pattern 52b can be a straight horizontal line (e.g., without having a modulation) as shown and the distance. The exemplary distances from surfaces 50a and 50c, may not match the reference arm path length, and thus interference patterns 52a, 52c having sinusoidal shapes may result.

Short Time Fourier transform ("STFT") or other Space-time or frequency-time signal transforms including, e.g., the Wigner transform, pseudo-Wigner transform, wavelet transforms, Fractional Fourier Transform, etc. of the CCD line scan signal can facilitate the determination of the axial information of the sample. For example, a strong reflection from the first surface of an object can usually occur. Therefore, the intensity of the highest peak in each Fourier transform may be proportional to the intensity of the reflection (assuming no DC term), and the location of that peak (which corresponds to the frequency) is proportional to the depth (z) of that surface. Reflections from other depths (for example, from locations under the tront surtacej, may manifest in different, often lower in intensity, locations in the Fourier trace. As a result, sub-surface imaging may be performed by determining the intensity of the Fourier component adjacent to the strongest peak.

In the exemplary graphs and images of FIG. 2, with the assumption that the surface reflection is higher than any reflections within the sample, the frequency at which the STFT amplitude is a maximum may likely be proportional to the step height intensity.

Since the spectrum detected by the linear array can provide likely only the real value of the complex field, however, it may be difficult to resolve the ambiguity of whether the surface height is above or below the "zero plane" with a single spectral measurement. This ambiguity in axial location is a known characteristic of SD-OCT (and optical frequency domain interferomerry - "OFDI"), where depth data wrapping may occur around the zero path difference between the reference and the sample arms. One exemplary solution to this problem can be to place the sample only in one- half of the measurement range. This exemplary method may be disadvantageous since it can decrease the ranging depth by a factor of two.

Several exemplary techniques for recovering the complex field and removing the depth ambiguity have been demonstrated for SD-OCT, including, e.g., the use of a 3x3 fiber-splitter and by acquiring several A-lines at slightly different reference arm path lengths.

FIGS. 3A and 3B and FIGS. 3C-3K show are a series of images and graphs, respectively, which indicate a use of an exemplary embodiment of a technique for extracting three-dimensional ("3D") data from a raw (e.g., an unprocessed; image. For example, FIG. 3A shows an exemplary raw image. Inset with a white-light image being of the face of a doll. FIG. 3B shows an expanded region encased in a square in FIG. 3A. FIGS. 3C-E show an exemplary graph of the intensity cross-sections provided along the solid and the dashed lines at the three locations marked in FIG. 3B. FIGS. 3F-3H show exemplary graphs of the corresponding power spectra of FIGS. 3C-3E. FIGS. 3I-3K show exemplary graphs of the corresponding cross- correlation between the power spectra of adjacent (solid and dotted) lines.

Referring to FIGS. 3A-3K in further detail, to resolve the depth ambiguity in SD-SEE, it is possible to acquire two spectra at different reference arm path lengths.

Comparing the phases of such STFT' s can facilitate a determination of whether a surface location is above or below a "zero plane." However, due to difficulties in accurately determining the phase in the presence of noise, such exemplary technique can have a high error rate when imaging tissue, resulting in a noisy three-dimensional reconstruction.

To. overcome the above-described problem, it is possible to employ an exemplary technique to determine the cross-correlation between immediately adjacent sample locations, while the reference arm is scanned at a slow rate was implemented. The sign of the offset of the first cross-correlation maximum is utilized to remove the height ambiguity. The implementation of this exemplary cross-correlation method can be accomplished by, e.g., stepping the sample arm slow scan (y-axis) to one location, acquiring one spectral scan, stepping the reference arm path length and acquiring an additional spectral scan. This exemplary procedure may be repeated for the entire volumetric image. Moving either the sample arm or the reference path (KSUD galvanometer) m small steps can, however, be difficult at high speeds. As a result, the reference arm can be continuously scanned at a slow rate (30 Hz, sawtooth waveform, typically 100 μm amplitude). The sample arm can also be continuously scanned, and spectral data can be over-sampled by capturing 5 spectrally-encoded lines per resolution element (which may be determined by the numerical aperture of the SEE probe lens).

Turning to FIG. 3 A, an exemplary raw image obtained from a face of a small plastic doll is shown therein. A white-light image of the doll's face is illustrated in the inset for reference. FIG. 3B shows an image of an expanded region of the raw image, displaying the interferometric fringes in more detail. In an exemplary embodiment, the detection spectrometer can measure a spectral resolution of approximately 0.1 ran, which is about 20 times higher than the spectral resolution at the imaging probe. In the interference pattern shown in Fig. 3 A, areas that are lower (or higher) than the zero plane have a diagonal flow-like pattern from top-right (top- left) to the bottom-left (bottom-right).

The STFT of each horizontal line of the raw image can be determined, e.g., using 250 rectangle windows with each window being 32 pixels wide. The data processing graphs associated with the procedure are illustrated in FIGS. 3C-3K for the three sample locations shown in FIG. 3B, respectively. The intensity cross- section along the 32-pixel-wide window pairs, indicated as solid and dashed lines on FIG. 3B, are shown in solid and dashed curves, respectively, on the three respective graphs of FIGS. 3C-3E. The corresponding exemplary intensity graphs of the FT's are shown in solid and dashed lines in FIGS. 3F - 3H. The exemplary graphs of the cross-correlation between the solid and the dashed curves within each plot are shown in FIGS. 31 - 3K.

For example, a full recovery of the 3D image can be obtained by performing these operations for every resolution element on the sample. An exemplary image which maps the scattering intensity from the doll's face can be obtained by determining the logarithmic of the maximum intensity value of each FT, and is shown in FIG. 4A. Such exemplary 2D reflectance image of the doll's face indicates regions of a low reflectance, such as the eyes and the eyebrows. The characteristic speckle pattern typical of coherent imaging is also present in this exemplary image.

FIG. 4B shows an image of an exemplary height map of the doll's face. For example, the depth dimension in this exemplary image is "folded" around the zero- plane (the doll's nose appears to have the same height as the lower lip). The signs of the locations of the maximum values of each cross-correlation may form the unwrapping sign mask, as shown in the exemplary image of FIG. 4C. This exemplary image can be used to unwrap the depth dimension, e.g., by multiplying it with the height map (FIG. 4B). The resulting exemplary unwrapped height map image, shown in FIG. 4D, may have two times greater depth range (e.g., 20 resolvable points) than the unwrapped image (see FIG. 4B, 10 axial points). Since the height can be determined for every point in the image, the height map may not be negatively effected by speckle noise, such as seen on the 2D image of FIG. 4 A. This exemplary technique for unwrapping the depth dimension may also not be negatively effected by losses involved with using an additional fiber coupler, and due to the continuous scanning of the RSOD, may not need any decrease in the imaging speed. Compared with time-domain SEE that was limited by the scanning range o± the RS UD [l.b mm), here the depth range was approximately 3 mm, limited by the spectrometer resolution.

Exemplary images generated by real-time 3D imaging techniques for various types of samples, including, e.g., a small paper flower, a skin fold on a volunteer's hand, and the tail between the hind paws of a mouse embryo, are shown in FIGS. 5A- 5C. For example, FIGS. 5A-5C are a series of transverse 2D images, depth images, and depth superimposed on the 2D image (shown in the left images 510, 540, 570, center images 520, 550, 580 and right images 530, 560, 590, respectively), and presented with the same scale. The left images 510, 540, 570 of each of FIGS. 5A-5C illustrate the 2D images. The center images 520, 550, 580 of FIGS. 5A-5C show the surface height encoded in gray levels, where high pixel value represent surface height closer to the imaging probe. The right images 530, 560, 590 of FIGS. 5A-5C show the depth information superimposed on the 2D image, where height is encoded with color. The exemplary images can be acquired at a rate of about 30 frames per second. The SNR can be 35dB, 3OdB and 25dB for images in FIGS. 5A, 5B and 5C, respectively.

In particular, FIG. 5A illustrates the images 510, 520, 530 with the depth range of the flower with can cover about 2.2 mm. Certain fine details are provided in the ID image of the skin fold, as shown in the left image 540 of FIG. 5B. The height map image 550 as shown in the center image of FIG. 5B, illustrates a large height difference between the right image 540 and the left image 560 of FIG. 5B. For example, to simulate endoscopic imaging of a fetus, a mouse embryo can be imaged through an approximately 2 mm thick plastic wall of a polypropylene 50 ml tube, and through approximately 10 mm thick layer of 3.7% formaldehyde fixative solution. FIG. 5C shows exemplary three-dimensional ("3D") images 560, 5 /U₅ 5bU oi two hind paws and a tail of a mouse embryo. Transverse 2D image, depth image, and depth superimposed on the 2D image are shown in the left image 560, the center image 570 and the right image 580 of FIG. 5C, respectively, and presented on the same scale. A significant amount of information present in the depth image in the center image 570 of FIG. 5C, compared to the 2D image provided in the left image

580 of FIG. 5C. While the exemplary 2D image of one of the mouse's hind paws reveals only three fingers, the depth image illustrates the 4th finger, as well as the paw's structure. The depth resolved image 570 also shows the relative location in space of the tail and the second paw.

FIG. 6 shows a flow diagram of an exemplary procedure for providing the three-dimensional ("3D") data reconstruction according to one exemplary embodiment of the present invention. For example, after illuminating a transverse line (step 610), two spectra can be captured in steps 615 and 625 with a small difference in reference path of approximately λ/4 (step 620). By determining the peak value of the short-time Fourier transform of the captured spectra in steps 630 and 635, the reflectance values of the entire transverse line can be obtained (step 640). The Fourier transform peak location can provide information on the axial distance in step 645 from the reference plane 50b. Unwrapping the axial dimension can be obtained by determining the short-time cross-correlation between the two spectra (step 650), determining the location of the first maxima (step 655), and using that information to determined if the measured location is above or below the reference plane (step 660). In this exemplary manner, the location height can be obtained in step 670. When the transverse and axial dimensions are determined, another lme on the sample can oe imaged (step 680).

SNR analysis

Exemplary SEE techniques can be performed using a CCD-based spectrometer that may record back reflections from the sample. Without a reference arm, this approach can be used only for imaging in 2D. When the reflectivity from the sample is low, which may be often the case for biomedical imaging, the electrical noise of the CCD camera can be the dominant noise source. Assuming a uniformly flat spectrum, the SNR can be given by the following:

SNΛ _(γ)

in which:

η denotes the quantum efficiency including the spectrometer efficiency;

P_s denotes the total sample power;

n_noi_se denotes the number of noise electrons of a single CCD pixel;

"_stgnai - ηKP.τ /(N_xHv) corresponds to the number of single electrons per

pixel associated with the sample light; and

nphotons ⁼ JJ-

With n_noise=nθ, N_x = 100, N₂ = 10, τ = 1 ms, P_s = 1 mW, and η = 0.5, the exemplary minimum detectable reflectivity corresponding to SNR2D=1 is R = 7.7e-9 at 800 nm wavelength. In time-domain SEE, the light reflected from the sample can be combined witn the reference light in a Michelson interferometer employing a scanning delay line. An

STFT of the interference signal can be recorded with a photodiode, thus producing a

3D image. Assuming a shot-noise limited detection, the SNR associated with a particular spatial point with reflectivity R can be given by the following:

⁽²⁾

in which:

P_r denotes the total reference power; P_s denotes the total reference power;

B = Nj[2τ) denotes the measurement bandwidth

τ denotes the integration time (line scan period);

v denotes the optical frequency;

N_x denotes the number of transverse resolvable points per spectrally- encoded line;

N_z denotes the number of axial resolvable points per spectrally- encoded line;

The SNR can be inversely proportional to the square of the number of transverse resolvable points since only a fraction of the reference arm power (P_r/N_x) interferes with the light returning from a single transverse location. With N_x = 100, N₂ = 10, τ = 1 ms, and P_s = 1 mW, the minimum detectable reflectivity is determined to be R = 1.25e-8 at 800 nm. Typical SΝR for imaging tissue was 6-10 dB. For the exemplary SD-SEE techniques, at least some oi wtacn are αescnoeα herein, the spectrometer can contains N_X*N_Z pixels. The exemplary SNR can be given by the following:

in which:

5 = l/(2r) ; and

τ is the acquisition time of each horizontal line.

With N_x = 100 and N₂ = 10, the exemplary minimum detectable reflectivity can be R = 1.25e-5 at 800 an, e.g., three (3) orders of magnitude more sensitive than the direct detection 2D SEE and the time-domain SEE. The typical SNR for the tissue imaging which can be obtained using the exemplary techniques according to certain exemplary embodiments of the present invention may be about 30-35 dB.

The exemplary SD-SEE techniques provide a significant improvement in spectrally-encoded imaging. In comparison with the conventional techniques, the exemplary SD-SEE techniques facilitate 3D imaging at, e.g., two orders of magnitude higher SNR, more than six-fold increase in the imaging speed, two-fold increase in the depth range. Due to the higher SNR and a preference of only a slow scanning of the reference arm (e.g., for depth unwrapping), the imaging speed may be limited only by the camera's speed, if at all. As an alternative or in addition, the high SNR can facilitate the use of imaging probes with low numerical aperture and long working distance, this allowing a large field of view for applications to be obtained, where imaging a large surface area is desired. The actual benefit of 3D imaging is mamly application dependent and remaiiib to be studied in the clinic. In the images shown in FIGS. 4A-4D, besides providing a better sense of orientation in space, the addition of the depth information may significantly improve the resultant images. When the magnitude of the back reflection from the tissue surface is uniform and can have less than optimum details, the speckle-free depth map may resolve the details which may not be otherwise seen in the 2D image. The depth image 570 of the mouse hind paw of FIG. 5C is one such example.

An exemplary embodiment of the SEE technique according to the present invention using spectral-domain interferometry has been described above. For example, by using a high-speed line camera in a high-resolution spectrometer, each x- z plane can be captured in a single shot. This exemplary technique can allow the use of high-speed imaging with 3 order of magnitude improvement in SNR improvement over the conventional techniques. In one exemplary embodiment, the real-time 3D imaging of tissue can be performed at a rate of 30 frames per second, with SNR at about 30 dB.

The foregoing merely illustrates the principles of the invention. Various modifications and alterations to the described embodiments will be apparent to those skilled in the art in view of the teachings herein. Indeed, the arrangements, systems and methods according to the exemplary embodiments of the present invention can be used with and/or implement any OCT system, OFDI system, SD-OCT system or other imaging systems, and for example with those described in International Patent Application PCT/US2004/029148, filed September 8, 2004, U.S. Patent Application No. 11/266,779, filed November 2, 2005, and U.S. Patent Application No. 10/501,276, filed July 9, 2004, the disclosures ot whicn are mcorporaieu oy reiercuoc herein in their entireties. It will thus be appreciated that those skilled in the art will be able to devise numerous systems, arrangements and methods which, although not explicitly shown or described herein, embody the principles of the invention and are thus within the spirit and scope of the present invention. In addition, to the extent that the prior art knowledge has not been explicitly incorporated by reference herein above, it is explicitly being incorporated herein in its entirety. All publications referenced herein above are incorporated herein by reference in their entireties.

Claims

What Is Claimed Is:

1. A system comprising: at least one first arrangement configured to provide a particular radiation which includes at least one first electro-magnetic radiation directed to at least one sample and at least one second electro-magnetic radiation directed to a reference arrangement, wherein at least one of the at least one first radiation or the at least one second radiation comprises a plurality of wavelengths, and wherein the at least one first arrangement is configured to spectrally disperse the at least one first electro-magnetic radiation along at least one portion of the at least one sample, and wherein the at least one second electro-magnetic radiation is measured at at least two different lengths of the reference arrangement with respect to the at least one first arrangement; and at least one second arrangement comprising a spectrometer arrangement which is configured to generate data associated with the at least one first electro-magnetic radiation and the at least one second electro-magnetic radiation obtained at the at least two different lengths.

2. The system according to claim 1, wherein the reference arrangement comprises at least one of a translatable mirror arrangement, a piezo-electric fiber stretching arrangement, a pulse-shaping arrangement, a rapidly-scanning optical delay line arrangement or an electro-optical or acousto-optical arrangement.

3. The system according to claim 1, wherein the second arrangement generates the data based on a Fourier transform of information received by the spectrometer arrangement which is associated with the at least one first electro-magnetic ramanon and the at least one second electro-magnetic radiation.

4. The system according to claim 3, wherein the data is generated as a function of a phase of the information.

5. The system according to claim 1, wherein the second arrangement generates the data based on at least one of a time frequency transform or a space-frequency transform of information received by the spectrometer arrangement which is associated with the at least one first electro-magnetic radiation and the at least one second electro-magnetic radiation.

6. The system according to claim 5, wherein the transform is a Short-time Fourier transform.

7. The system according to claim 1, wherein the second arrangement generates the data based on a correlation between information received at at least one first length of the at least two different lengths and at least second length of the at least two different lengths.

8. The system according to claim 7, wherein the correlation is a cross- correlation.

9. The system according to claim 8, wherein the second arrangement determines a first peak of the cross-correlation, and determines a sign of the cross-correlation based on further information associated with the first peak.

10. The system according to claim 1, wherein the second arrangement generates the data based on a comparison of phases between information received at at least one first length of the at least two different lengths and at least second length of the at least two different lengths.

11. The system according to claim 10, wherein the second arrangement generates further data based on a magnitude of a Fourier transform of the data.

12. The system according to claim 11, wherein the second arrangement combines the data and the further data to form a composite image data associated with the at least one sample.

13. The system according to claim 1, wherein the data is associated with at least one portion of the at least one sample which is located in a direction that is axial with respect to a direction of the at least one first electro-magnetic radiation.

14. The system according to claim 9, wherein the data is associated with at least one portion of the at least one sample which is located in a direction that is axial with respect to a direction of the at least one first electro-magnetic radiation.

15. The system according to claim 14, wherein the sign ot a location or me πrsi peak of the cross-correlation is associated with the at least at least one portion of the at least one sample which is located in the direction that is axial with respect to the direction of the at least one first electro-magnetic radiation.

16. The system according to claim 1, wherein the data is further associated with at least one of a two-dimensional image or a three-dimensional image of at least a portion of the at least one sample.

17. The system according to claim 1, wherein the at least one second electromagnetic radiation is directed to continuously scan the different lengths of the reference arrangement.

18. A method comprising: providing a particular radiation using at least one first arrangement, the at least one particular radiation including at least one first electro-magnetic radiation directed to at least one sample and at least one second electro-magnetic radiation directed to a reference arrangement, wherein at least one of the at least one first radiation or the at least one second radiation comprises a plurality of wavelengths; spectrally dispersing the at least one first electro-magnetic radiation along at least one portion of the at least one sample, and wherein the at least one second electro-magnetic radiation is measured at at least two different lengths of the reference arrangement with respect to the at least one first arrangement; and generating data associated with the at least one first electro-magnetic radiation and the at least one second electro-magnetic radiation obtained at the at least two different lengths using at least one second arrangement whicn comprises a spectrometer arrangement.

19. The method according to claim 18, wherein the data is generated based on a comparison of phases between information received at at least one first length of the at least two different lengths and at least second length of the at least two different lengths.

20. The method according to claim 18, further comprising combining the data and the further data to form a composite image data associated with the at least one sample.