US 7,365,859 B2Grant

System and method for optical coherence imaging

Issue Date:2008-04-29

•60 Claims

•16 Drawing Sheets

Abstract

A system and method for imaging of a sample, e.g., biological sample, are provided. In particular, at least one source electro-magnetic radiation forwarded to the sample and a reference may be generated. A plurality of detectors may be used, at least one of the detectors capable of detecting a signal associated with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference. At least one particular detector may have a particular electrical integration time, and can receive at least a portion of the signal for a time duration which has a first portion with a first power level greater than a predetermined threshold and a second portion immediately preceding or following the first portion. The second portion may have a second power level that is less than the predetermined threshold, and extends for a time period which may be, e.g., approximately more than 10% of the particular electrical integration time.

Metadata

Assignee

The General Hospital Corporation

Inventors

Seok-Hyun Yun
Brett Eugene Bouma
Guillermo J. Tearney
Johannes F. deBoer

Application Information

Application Number:US 11/225,840

Filing Date:2005-09-12

Priority Date:2004-09-10

Art Unit:2886

Classifications

IPC:

G01B11/02

Patent Drawings (16 sheets)

Description

Cross-Reference to Related Application(s)

[0001] The present invention claims priority from U.S. Patent Application Ser. No. 60/608,800 filed on Sep. 10, 2004, the entire disclosure of which incorporated herein by reference.

Field of the Invention

[0002] The present invention relates generally to optical coherence tomography imaging, and more particularly, to a system and method that uses optical coherence tomography that permits imaging of biological samples with high sensitivity and reduced artifacts, e.g., due to sample and probe motion.

Background of the Invention

[0003] Image artifacts resulting from motion have been important issues of research in many medical imaging modalities because they may degrade the image quality and cause inaccurate clinical interpretation of images. Artifacts can arise when an object being imaged (sample) is moved during data acquisition but is assumed stationary in the image reconstruction process. In each imaging modality, motion artifacts can be present in different forms and with different magnitudes. Understanding basic motion effects in a particular imaging method is an essential step toward the development of techniques to avoid or compensate resulting artifacts. Optical interferometric imaging methods using frequency domain ranging have recently received considerable interest due to their high image acquisition speed and sensitivity.

[0004] Two frequency domain techniques have been demonstrated: spectral-domain optical coherence tomography (SD-OCT) as described in A. F. Fercher et al., “Measurements of intraocular distances by backscattering spectral interferometry,” Opt. Comm. 117, 43-48 (1995), G. Hausler et al., “Coherence radar and spectral radar—new tools for dermatological diagnosis,” J. Biomed. Opt. 3, 21-31 (1998), M. Wojtkowski et al., “Real time in vivo imaging by high-speed spectral optical coherence tomography,” Opt. Lett. 28, 1745-1747 (2003), N. Nassif et al., “In-vivo human retinal imaging by ultra high-speed spectral domain optical coherence tomography,” Opt. Lett. 29, 480-482 (2004), S. H. Yun et al., “High-speed spectral domain optical coherence tomography at 1.3 μm wavelength,” Opt. Express 11, 3598-3604 (2003), and optical frequency domain imaging (“OFDI”) S. R. Chinn, E. Swanson, and J. G. Fujimoto, “Optical coherence tomography using a frequency-tunable optical source,” Opt. Lett. 22, 340-342 (1997), B. Golubovic et al., “Optical frequency-domain reflectometry using rapid wavelength tuning of a Cr⁴⁺:forsterite laser,” Opt. Lett. 22, 1704-1706 (1997), F. Lexer et al., “Wavelength-tuning interferometry of intraocular distances,” Appl. Opt. 36, 6548-6553 (1997), S. H. Yun et al, “High-speed optical frequency-domain imaging,” Opt. Express 11, 2953-2963 (2003), the entire disclosures of all of which are incorporated herein by reference. Using the SD-OCT technique, the spectral interference fringe can be measured in the spatial domain by means of a diffraction grating and a charge-coupled device (“CCD”) array. In exemplary OFDI techniques, the spectral fringe is mapped to the time domain by use of a frequency-swept light source and measured with a photodetector as a function of time. In both methods; axial reflectance profile (A-line) is obtained by performing a discrete Fourier transform of the acquired data. Since the Fourier transform process involves integration of the entire data set obtained in single A-line period, the signal-to-noise ratio (“SNR”) is enhanced relative to time domain ranging, as described in S. H. Yun et al., “High-speed optical frequency-domain imaging,” Opt. Express 11, 2953-2963 (2003), R. Leitgeb, et al. “Performance of Fourier domain vs. time domain optical coherence tomography,” Opt. Express 11, 889-894 (2003), J. F. de Boer et al., “Improved signal-to-noise ratio in spectral-domain compared with time-domain optical coherence tomography,” Opt. Lett. 28, 2067-2069 (2003), and M. A. Choma et al., “Sensitivity advantage of swept source and Fourier domain optical coherence tomography,” Opt. Express 11, 2183-2189 (2003), the entire disclosures of all of which are incorporated herein by reference This improvement in SNR is particularly advantageous for applications requiring high image acquisition rates such as screening for disease and surveillance of large tissue volumes. It is, however, possible that the integration effect enhances the sensitivity to sample motion because the motion-induced change in signal is also integrated over the entire A-line acquisition period.

[0005] Spectral-domain optical coherence tomography (“SD-OCT”) makes use of low-coherence spectral interferometry to obtain cross-sectional images of a biological sample. Interference fringes as a function of wavelength are measured using a broadband light source and a spectrometer based on a charge-coupled-device (“CCD”) camera. The axial reflectivity profile of a sample, or an A-line, can be obtained by a discrete Fourier transform of the camera readout data. This imaging technique has recently gone through rapid technical development to demonstrate high quality imaging of biological samples with fast image acquisition time, an order of magnitude faster than state-of-the-art time-domain OCT systems. The recent advancement in imaging speed may lead to the utilization of SD-OCT in a number of clinical applications in the near future.

[0006] The SD-OCT systems that have been used to date utilized either a continuous-wave (“cw”) broad-spectrum light source, such as super luminescent diodes (“SLD”), or ultrashort mode-locked pulses with a high repetition rate in the range of 10-100 MHz. In both cases, the CCD array is generally illuminated constantly, and therefore the exposure time of the CCD camera determines the signal acquisition time for a single A-line. In this case, a path length change in the interferometer during image acquisition results in phase drift in the interference fringe. If the phase drifts over more than μ during a single A-line acquisition, the interference fringe can be completely erased, resulting in a degradation of SNR. This motion artifact can be caused by axial motion of a sample relative to the probe beam. By comparison, transverse sample motion or transverse beam scanning does not result in fringe washout. However, the transverse motion can result in degradation in transverse resolution and SNR. In medical imaging in vivo, the motion effects can arise from various sources. The main causes include patient motion, physiological phenomena such as cardiac motion, blood flow, pulsation, and catheter movement associated with beam scanning or uncontrolled movement of operator's hand. Furthermore, environmental changes such as mechanical vibration, sound waves, and temperature drift can alter the path length difference in the interferometer, resulting in SNR degradation through fringe washout. Considering that cameras appropriate for SD-OCT typically provide exposures times longer than 10 μs, a solution to the fringe washout problem will be required for biomedical applications where sample and probe motion is common.

[0007] Therefore, one of the objects of the present invention is to reduce or eliminate the motion artifacts.

Summary of the Invention

[0008] According to the present invention, an imaging apparatus/system is provided which includes an optical source and at least one detector array. In one exemplary embodiment of the present invention, an optical source can emit a broadband spectrum in a pulsed mode, for example, by Q-switching or mode locking, with a pulse repetition rate preferably being equal to a readout rate of a detector array. The pulsed source can produce enough average optical power to provide sufficient signal to noise ratio required for imaging, while the relatively short duration of the output pulses results in an effective signal integration time substantially shorter than the detector's integration time, leading to high-sensitivity motion-artifact-free imaging. This pulsed-source approach may pertain to full-field optical coherence tomography and/or spectral-domain optical coherence tomography. In another exemplary embodiment of the present invention, the optical source is a wavelength-swept source emitting relatively narrowband spectrum swept over a wide range with a repetition rate preferably being equal to the readout rate of the detector array or A-line rate. This exemplary embodiment of the present invention allows the interference signal associated with each spectral component to be measured with an effective integration time substantially shorter than an A-line acquisition time. This exemplary scheme may also eliminate the fringe washout problem as in the prior art using continuous-wave broadband source or high-repetition mode-locked pulses. The above-described exemplary embodiments of the present invention may employ two or more detector arrays for dual-balanced detection and/or polarization diversity and further employ fiber-optic probes, allowing for medical imaging in vivo with high sensitivity, high speed, and the immunity from motion artifacts.

[0009] Accordingly, an exemplary embodiment of a system and method for imaging at least a portion of a sample are provided. In particular, at least one source electro-magnetic radiation can be generated and forwarded to the sample and a reference. A signal associated with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference can be detected using at least one of a plurality of detectors. At least one particular detector can have a particular electrical integration time. Such detector may received at least a portion of the signal for a time duration which has at least one first portion with at least one first power level that is greater than a predetermined threshold and at least one second portion immediately preceding or following the at least one first portion. The second portion can have at least one second power level which is less than the predetermined threshold, and may be extended for a time period which is approximately at least 10% of the particular electrical integration time.

[0010] In addition, the signal may be at least one of frequency components of the combination, and the particular detector can receive such frequency component. The source electro-magnetic radiation can be generated by a source arrangement which may be a pulsed broadband source. The source electro-magnetic radiation generated by the pulsed source may be a single pulse per the particular electrical integration time. The pulsed source may be a Q-switched laser, a cavity-dumped mode-lock laser, and/or a gain-switched laser. The source electro-magnetic radiation generated by the source arrangement may be a burst of radiation that extends for at most approximately 90% of the particular electrical integration time. The burst of radiation may include multiple pulses. The source electro-magnetic radiation generated by the pulsed broadband source can have a spectrum with (i) a center wavelength between approximately 700 nanometers and 2000 nanometers, and/or (ii) a spectral width of approximately greater than 1% of the center wavelength. The source electro-magnetic radiation generated by the pulsed broadband source may have a pulse width approximately shorter than 1 μsec. A duration of the burst of radiation can be approximately shorter than 1 μsec.

[0011] According to another exemplary embodiment of the present invention, the source arrangement generating the source electro-magnetic radiation may include an optical gating switch. A frequency of the source electro-magnetic radiation can vary over time. A mean frequency of the source electro-magnetic radiation may change (i) substantially continuously over time at a tuning speed that is greater than 100 terahertz per millisecond, and/or (ii) with a repetition period that is less than approximately 90% of the particular electrical integration time. The source electro-magnetic radiation can have a tuning range (i) with a center wavelength between approximately 700 nanometers and 2000 nanometers, and/or (ii) of approximately greater than 1% of the center wavelength. The source electro-magnetic radiation may have an instantaneous line width and a tuning range, with the instantaneous line width being less than approximately 10% of the tuning range. The source arrangement may include (i)a tunable laser, (ii) a tunable filter, and/or (iii) a medium, and can generate the source electro-magnetic radiation based on a non-linearity associated with the medium. The frequency may vary substantially (i) linearly with time, and/or (ii) sinusoidally with time.

[0012] A detector arrangement which includes the detectors can be provided, that includes an electrical shutter that is adapted to gate a transmission of photoelectrons associated with the combination of the first and second electro-magnetic radiation, wherein a time period for the gating to allow the transmission of the photoelectrons is less than approximately 90% of the particular electrical integration time. The sample can be a biological sample. The detection arrangement may include at least one charged-coupled device. The source arrangement may be a pulsed broadband source. At least one spectral separating unit can be provided which separates spectrum of the first electro-magnetic radiation, the second electro-magnetic radiation and/or the combination into the at least one of the frequency components.

[0013] These and other objects, 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

[0014] Further objects, features and advantages of the invention will become apparent from the following detailed description taken in conjunction with the accompanying figures showing illustrative embodiments of the invention, in which:

[0015] FIG. 1 is an exemplary schematic diagram of a conventional SD OCT system;

[0016] FIG. 2A is a set of exemplary graphs of spectral and temporal characteristics obtained from a conventional continuous-wave optical source;

[0017] FIG. 2B is a set of exemplary graphs of spectral and temporal characteristics obtained from a high-repetition rate mode locked laser source;

[0018] FIG. 3 is a set of exemplary graphs of spectrum and temporal characteristics obtained from a low-repetition rate broadband source;

[0019] FIGS. 4(a)-(c) are block diagrams of exemplary embodiments of low-repetition broadband source arrangements according to the present invention;

[0020] FIGS. 5(a)-(d) are exemplary graphs of spectrum and temporal characteristics obtained from an exemplary wavelength-swept source;

[0021] FIGS. 6(a)-(c) are block diagrams of exemplary embodiments of an exemplary wavelength-swept source arrangements according to the present invention;

[0022] FIG. 7(a) is an exemplary illustration of a detection signal of a cw light with a CCD array in a spectrometer;

[0023] FIG. 7(b) is an exemplary illustration of a detection signal of a pulsed light with a CCD array in a spectrometer;

[0024] FIG. 7(c) is an exemplary illustration of a detection signal of a swept light with a CCD array in a spectrometer;

[0025] FIG. 8(a) is a schematic of an exemplary pulsed ASE source;

[0026] FIG. 8(b) is a schematic of an exemplary wavelength-swept source;

[0027] FIGS. 9(a) and (b) are illustrations of signals of exemplary temporal and spectral output characteristics obtained from the exemplary pulsed ASE source;

[0028] FIGS. 9(c) and (d) are illustrations of signals of exemplary temporal and spectral output characteristics obtained from the exemplary swept source;

[0029] FIG. 10 is an exemplary block diagram of an exemplary embodiment of an SD OCT system according to the present invention;

[0030] FIG. 11 is an exemplary illustration of SD-OCT images of a paper, acquired when a sample is static (in sections a, c and e) and moving at 80 Hz over 0.8 mm (in sections b, d and f) with three different light sources;

[0031] FIG. 12(a) is an illustration of a variation of a total signal power, a sum of reflectivity of 256 depth points in each A-line, as a function of A-line index or time, obtained from images a and b shown in FIG. 11;

[0032] FIG. 12(b) is an illustration of a variation of a total signal power, a sum of reflectivity of 256 depth points in each A-line, as a function of A-line index or time, obtained from images c and d shown in FIG. 11;

[0033] FIG. 12(c) is an illustration of a variation of a total signal power, a sum of reflectivity of 256 depth points in each A-line, as a function of A-line index or time, obtained from images e and f shown in FIG. 11;

[0034] FIG. 13 is a set of illustrations of exemplary SD-OCT images of a human. coronary artery in vitro, obtained using an exemplary embodiment of the system according to the present invention, with a rotation speed of a catheter and the light source (Section A—4.5 rps, cw ASE source, Section B—37.9 rps, cw ASE source, Section C—4.5 rps, swept source, and Section C—37.9 rps, swept source);

[0035] FIG. 14 is a block diagram of another exemplary SD-OCT system that uses a line pulsed or swept source and a two-dimensional CCD array; and

[0036] FIG. 15 is a block diagram of an exemplary full-field OCT system that uses a pulsed source and a two-dimensional CCD array.

Detailed Description

[0037] FIG. 1 depicts an exemplary basic configuration of a spectral-domain optical coherence tomography (“SD-OCT”) system. Broadband light 10 is split by a coupler 20 into a sample arm 22 and a reference arm 24 that is terminated by a mirror 26 at its distal end. A probe 30 at the end of the sample arm delivers light to a sample 40, and receives the light backscattered from within the sample. The light returned from the two interferometer arms is recombined and directed via a circulator 44 to a spectrometer 50 consisting of a collimator 52, a diffraction grating 54, and a lens 56, a CCD array 60, and camera 62. Individual pixels of the CCD array 60 measure the optical power as a function of wave number, k=2π/λ where λ is the optical wavelength. The CCD output is digitized using a digitizer 70 and processed in a computer 74. A discrete Fourier transform (“DFT”) of the CCD scan output produces an axial reflectance profile of the sample (A-line). A 2-D tomographic image can be obtained by acquiring multiple A-lines as the probe beam is scanned over the sample along a transverse direction. This exemplary architecture and the operating principle described above are well known in the art.

[0038] The broadband optical source used in prior art can be categorized into two types: continuous wave (“cw”) as shown in FIG. 2A, and mode-locked pulsed source as shown in FIG. 2B. The cw source emits constant spectrum and constant output power. In this case, the integration time of OCT signals is equal to the exposure time of the detector array. Examples of such cw sources include super luminescent diodes, amplified spontaneous emission (“ASE”) source, and supercontinuum source. On the other hand, the mode-locked source emits very short optical pulses with a duration ranging from sub nanoseconds to several femtoseconds and a relatively high repetition rate from 10 MHz to 1 GHz. The exposure time of CCD is typically in the order of 10 microseconds to 10 milliseconds. As a result, the mode-locked pulses essentially behave like continuous wave, illuminating the CCD array constantly over its entire exposure time. The use of the cw and mode-locked source in OCT has two shortcomings: (a) significant motion artifacts due to relatively long signal integration time and (b) SNR degradation in case <100% duty cycle of signal integration in the detector array. The exemplary embodiments according to the present inventions provide sample solutions to such problems.

[0039] One exemplary embodiment of the present invention relates to a system for imaging of a sample, e.g., biological sample, which may include a source arrangement that generates at least one source electro-magnetic radiation forwarded to the sample and a reference. Such exemplary system may include at least one detection arrangement that has a plurality of detectors, at least one of the detectors capable of detecting a signal associated with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference. At least one particular detector may have a particular electrical integration time, and can receive at least a portion of the signal for a time duration which has a first portion with a first power level greater than a predetermined threshold and a second portion immediately preceding or following the first portion. The second portion may have a second power level that is less than the predetermined threshold, and extends for a time period which is approximately more than 10% of the particular electrical integration time.

[0040] The electro-magnetic radiation is preferably light with a center wavelength in the range of 700 to 2000 nm. The detector array is preferably charge-coupled devices (“CCD”). Using the exemplary SD-OCT system, the signal detected in the detector array is frequency components of the combination, or the spectrum. Typically the spectrum is obtained using a spectrally separating device such as a diffraction grating. A number of methods to obtain the spectrum with detector arrays are well known in the art. For full-field OCT, the signal is the optical power of the combination, which is linked to specific transverse locations in the sample.

[0041] In another exemplary embodiment of the present invention, the source arrangement can be a pulsed broadband source generating a single pulse per the particular electrical integration time or producing a burst of radiation that extends for at most approximately 90% of the particular electrical integration time. Each burst may include multiple ultrashort optical pulses in it. Examples of the pulsed sources include a Q-switched laser, a cavity-dumped mode-lock laser, and a gain-switched laser. Preferably, the spectrum of the pulsed source may have a spectral width of approximately greater than 1% of the center wavelength and a pulse width or a duration of the burst of radiation approximately shorter than 1 microseconds. The source arrangement may comprise a broadband cw source and an optical gating switch or electrical shutter integrated in the CCD array. The time window where the optical power is less than the threshold can be considered as OFF state, and the window where the power is greater than the threshold as ON state. The threshold is preferably less than 50% of the power level during ON state, however a typical pulsed source may provide much larger power extinction between the ON and OFF state. During a single detector integration time, one or multiple ON states may exist, however the total illumination span, or the duration from the start of the first ON state to the end of the last ON state is preferably shorter than 90% of the detector integration time. For example, the shorter the illumination span, the more suppression of motion artifacts can be obtained.

[0042] In yet another exemplary embodiment of the present invention, the source arrangement can be a wavelength swept source where a mean frequency of the output spectrum varies over time. The mean frequency of the source electro-magnetic radiation may change substantially continuously over time at a tuning speed that is greater than 100 terahertz per millisecond and repeatedly with a repetition period that is less than approximately 90% of the particular electrical integration time. The tuning range of the source electro-magnetic radiation may have a tuning range with a center wavelength between approximately 700 nanometers and 2000 nm, a tuning width of approximately greater than 1% of the center wavelength, and an instantaneous line width of less than approximately 10% of the tuning range. Such a source arrangement includes a tunable laser, soliton laser in conjunction with Raman self frequency shift, or cw broadband source in conjunction with a tunable filter. The mean frequency may vary substantially linearly or sinusoidally with time. As for the pulsed source, the time window where the optical power received by a specific pixel is less than the threshold can be considered as OFF state for the particular pixel, and the window where the power is greater than the threshold as ON state. The threshold is preferably less than 50% of the power level during ON state, however a typical pulsed source may provide much larger power extinction between the ON and OFF state. During the detector integration time of the pixel, one or multiple ON states may exist, however the total illumination span, or the duration from the start of the first ON state to the end of the last ON state is preferably shorter than 90% of the pixel integration time. The shorter the illumination span is, the more suppression of motion artifacts can be obtained.

[0043] According to still another exemplary embodiment of the present invention, a method may be provided for imaging of a sample, typically biological sample. For example, at least one source electro-magnetic radiation may be generated to be forwarded to the sample and a reference. At least a portion of a signal associated may be detected with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference using at least one detector of a plurality of detectors of a detection arrangement. At least one particular detector may have a particular electrical integration time, and can receive at least a portion of the signal for a time duration which has a first portion with a first power level greater than a predetermined threshold and a second portion immediately preceding or following the first portion. The second portion may have a second power level less than the predetermined threshold, and can extend for a time period which is approximately more than 10% of the particular electrical integration time.

[0044] FIG. 3 illustrates sample outputs of one exemplary embodiment of a system and method according to the present invention that is based on a broadband pulsed source. Such source emits a single burst of optical energy, or simply “pulse”, per each integration window of the detector array with timing synchronization between the pulses and integration window of the detector array, as illustrated in FIG. 3. Examples of such sources may include time-gated cw or mode-locked broadband source 200 using an external intensity modulator 210 (as shown in FIG. 4(a)), supercontinuum source based on a Q-switched pump laser 220 and supercontinuum generation medium 230 (as shown in FIG. 4(b)), self-Q-switched supercontinuum source or Raman source based on pump laser 240 and nonlinear medium 250 (as shown in FIG. 4(c)). The use of such source results in an effective signal integration time equal to the duration of the pulse, which may range from sub microseconds to sub nanoseconds, substantially shorter than the integration time of the detector array itself. Although a single pulse operation is described, other optical sources emitting multiple pulses per each integration time may be used if the pulses are generated within duration substantially shorter than the detector integration time. In exemplary clinical applications such as ophthalmology, the maximum optical energy or intensity level that can be illuminated to the retina is limited by potential damage to the tissue. Using the exemplary arrangement shown in FIG. 4(a), it is possible to use a broadband source emitting a high output power, such as SLD and Ti:Sapphire mode-locked laser, and time-gate the output to decrease the duty cycle and therefore an effective exposure energy level to the sample.

[0045] FIGS. 5(a)-(d) illustrate graphs to explain the principle of another exemplary embodiment of the present invention based on a wavelength-swept source which emits substantially narrowband spectrum that is swept over a wide spectral range, repeatedly in time. FIG. 5(c) show exemplary signals generated using a swept source with the output wavelength swept in a saw-tooth fashion. The tuning cycle is synchronized with the integration window of the detector array. In this exemplary case, each detector element can receive the light with corresponding wavelength during only a short period of time which determines the effective signal integration time. As a numerical example, when the total tuning range is 150 nm, centered at 1300 nm, and the instantaneous linewidth is 1 nm, the effective signal integration time can be only one hundredth of the detector integration time.

[0046] As shown in FIGS. 6(a)-(c), a wavelength-swept source may be implemented by using a conventional broadband source 300 followed by a wavelength scanning filter 310. According to an exemplary variant of the present invention, a wavelength-swept laser may be used using a gain medium 320, tunable filter 330 and output coupler 340 in a laser cavity 350. A wavelength-swept laser may be configured to yield a linewidth that is narrower than the resolution of the spectrometer; in this case the complexity and tolerance in spectrometer design may be relaxed. The combination of wavelength-swept source and detector array described above may be analogized with optical frequency domain imaging and exhibits motion artifacts such as Doppler distortion. To further reduce the motion artifacts, the wavelength-swept source may be operated in a low-duty-cycle or Q-switched regime, with an advantage of further reduction of effective signal integration time. Another possible source can includes a broadly tunable source based on soliton self frequency shift using a soliton source 360 and Raman medium 370.

[0047] Exemplary conventional SD-OCT systems utilize either a continuous-wave (cw) broad-spectrum light source, such as super luminescent diodes (SLD), or ultrashort mode-locked pulses with a high repetition rate in the range of 10-100 MHz. Full field OCT systems have typically employed cw thermal light source. For such conventional systems, the CCD array is illuminated constantly, and therefore the exposure time of the CCD camera determines the signal acquisition time for a single A-line. However, sample or probe motion during the A-line acquisition time can result in various undesirable artifacts such as signal fading and spatial resolution degradation. In particular, due to axial sample motion, the visibility of detected spectral fringes can diminish significantly resulting in significant image fading. Considering that cameras appropriate for SD-OCT typically provide exposures times longer than 10 μs, a solution to the fringe washout problem is preferable for biomedical applications where sample and probe motion is common.

[0048] FIGS. 7(a)-(c) illustrates exemplary illustration of a detection signal with a CCD array in a spectrometer how the signal detection in the exemplary SD-OCT system for three different light sources: broadband cw source (see FIG. 7(a)), broadband pulsed source (see FIG. 7(a)), and narrowband wavelength-swept source (see FIG. 7(a)). In this figure, spectrally dispersed, broad-spectrum light 400 is incident on a CCD array 410 so that each CCD pixel receives a narrowband portion of the source light. The vertical bars 420 represent the time window during which the camera integrates photon-generated electrons. FIG. 7(a) shows the signals obtained using a common implementation of the SD-OCT system. The operational principles of the systems generating signals shown in FIGS. 7(b) and (c) are described below.

[0049] In particular, FIG. 7(b) depicts a train of short broadband pulses 430 with a repetition rate equal to the CCD readout rate. The integration time of this exemplary system is given by the pulse duration rather than the camera readout time. As a result, snap-shot A-line profiles can be obtained with freedom from sample or probe motion. This exemplary technique is conceptually similar to the use of stroboscopic illumination in photography. Although for most biomedical applications nanosecond pulses are sufficiently short to avoid motion artifacts, it is interesting to note that in principle, this approach could provide femtosecond temporal resolution A-line acquisition through the use of low-repetition mode locked lasers. The following analysis, however, pertains to an arbitrary pulsed source delivering either single bursts of short-duration broadband light or bursts comprising a brief train of mode locked pulses.

[0050] To understand the imaging characteristics of a pulsed-source SD-OCT system, the signal-to-noise ratio (SNR) for pulsed and cw operation in the presence of axial motion may be reviewed. For example, let T_sand T_edenote the duration of the pulse and the electrical integration time of the camera, respectively. For a sample moving axially in parallel to an optical probe beam with a speed v_z, the signal power S, normalized to the signal at v_z=0, is given by
S≈|∫₀^T^eP(t)e^j2k⁰^v^z^tdt|²/∫₀^T^eP(t)dt², (1)
where P(t) represents time-varying optical power of the pulse, and k₀=2π/λ₀denotes the wave number corresponding to the center wavelength λ₀. Equation 1 yields S≈sin²(k₀Δz)/(k₀Δz)²for a square pulse and S≈exp[−k₀²Δz²/(2ln 2)]0 for a Gaussian pulse with T_sas the full-width-at-half-maximum (“FWHM”) pulse duration, where Δz=v_zT_srepresents the total sample movement during pulse duration T_s. These expressions imply that significant signal fading occurs if the sample movement is greater than a half optical wavelength during the pulse duration. Therefore, the short pulsed technique (T_s<<T_e) offers a significant advantage over the conventional cw operation in terms of motion-induced signal fading. Similarly, one can see that pulsed operation can also suppress other motion artifacts, such as spatial resolution degradation due to sample motion and transverse beam scanning.

[0051] The fundamental noise characteristics of pulsed operation are likely approximately identical to those of cw operation, because the detection bandwidth is solely determined by the integration time of the camera. If both a pulsed and cw sources produce the same average optical power and relative intensity noise (“RIN”), both would yield the same SNR in the limit of a stationary sample.

[0052] FIG. 7(c) shows signals generated using another exemplary techniques that uses another exemplary pulsed-source SD-OCT approach that is based on a narrowband, wavelength-swept source. Since the optical spectrum 440 incident to the CCD array 420 is continuously changed in time, each of the CCD pixels receives its corresponding spectral component only for a short time interval. As with pulsed broad bandwidth illumination, rapidly sweeping the wavelength allows the SD-OCT signal to be free from signal fading due to fringe washout. However, unlike pulsed operation, individual “spectrum pulses” do not arrive at the CCD pixels at the same time.

[0053] For a linear sweep shown in FIG. 7(c), the swept operation is approximately analogous to optical frequency domain imaging (“OFDI”) as described in U.S. patent application No. 60/514,769 filed Oct. 27, 2003, the entire disclosure of which is incorporated herein by reference. In this exemplary swept operation, spectral fringes are measured as a function of time using a swept source and a standard photodiode. Therefore, both imaging techniques can exhibit similar motion artifacts. The generation of motion artifacts in produced by the exemplary OFDI system is known. The swept-source operation in the exemplary SD-OCT system, however, differs from the operations of the OFDI system in that it does not require a linear tuning slope or narrow instantaneous linewidth of the source because these specifications are governed by the detection spectrometer. Such distinctions are significant considering that tuning speed and power in wavelength swept lasers are often limited by constraints on linearity and instantaneous linewidth.

[0054] For example, pulsed and wavelength-swept sources may be constructed according to an exemplary embodiment of the present invention. A block diagram of the exemplary system of the present invention which includes a gating device is shown in FIG. 8(a). The pulsed broadband source can be provided by an external time-gating of cw broadband amplified spontaneous emission (“ASE”) from a semiconductor optical amplifier 450 (e.g., SOA, Philips CQF 882/e). The output of the SOA, prior to the time gating, can be characterized as cw un-polarized ASE centered 1.3 μm, with 7-mW total power at an injection current of 450 mA. The cw ASE can be coupled to an external optical gating device which includes a polygonal mirror scanner 460 and lenses 462, 464, in conjunction with a circulator 466. The polygonal mirror had 40 facets with a facet-to-facet angle of 9 degrees. The focal lengths of the collimating 462 and focusing 464 lenses can be 11 and 100 mm, respectively, to obtain a duty cycle of approximately 5% in the output. FIG. 9(a) shows an output pulse train measured with an InGaAs photodetector and oscilloscope (detection bandwidth=100 MHz) as the polygon scanner may be rotated at 474 revolutions per second to produce a pulse repetition rate of 18.94 kHz. The measured pulse width and corresponding duty cycle were 2.85 μs (FWHM) and 5.4%, respectively. The average output power measured with a power meter can be 300 μW. FIG. 9(b) shows the output spectrum measured with an optical spectrum analyzer. The spectrum may be approximately identical to that of the input ASE, with a center wavelength at 1300 nm and a FWHM of 66 nm.

[0055] FIG. 8(b) shows a block diagram of another exemplary embodiment of a system according to the present invention which includes the wavelength-swept laser. The laser employed the same SOA 450 and a scanning wavelength filter based on a polygonal mirror scanner 460 in a fiber-optic ring laser cavity 468. The scanning filter consisted of a diffraction grating 470 (830 lines per mm), two lenses in 4f configuration (472; f=60 mm, 474; f=63.5 mm), and the same 40-facet polygonal mirror scanner 460 as used for the pulsed source. The scanning filter can be configured to have a free spectral range of 275 nm centered at 1320 nm wavelength, which may result in a duty cycle of the laser output closely matched to that of the CCD camera (46%). When the pass band of the filter scans outside the gain bandwidth of the SOA, the source likely does not reach the lasing threshold and simply produces ASE. FIG. 9(c) shows the temporal characteristics of the laser output at a sweep repetition rate of 18.94 kHz. The region where the output power varies with a Gaussian-like profile corresponds to when the source was operated above the lasing threshold. Outside this region, the output is ASE with a constant power. To determine how much the ASE level contributed to the detected light during swept laser operation, the backward-propagating ASE power was measured by inserting a 5% tap coupler in the cavity between the filter and SOA (lower trace in FIG. 9(c), gray line). The ASE level dropped significantly during laser operation because ASE was suppressed due to gain saturation in the SOA. The laser-to-ASE ratio reached as high as 16 dB in the middle of the lasing tuning range. Horizontal bars (green) represent the integration window of the camera, which was synchronized with laser tuning. The average output power measured with a power meter was 18 mW.

[0056] FIG. 9(d) shows an exemplary output spectrum measured with the optical spectrum analyzer in a peak-hold mode. In a peak-hold mode, the contribution of ASE to the measurement would be negligible owing to its much lower spectral density than laser light at a given time. Therefore, the measured spectrum represents the tuning envelope of the swept laser. The tuning range was approximately 135 nm, centered at 1325 nm. An exemplary instantaneous linewidth of the swept output was approximately 0.4 nm, as determined by measuring the coherence length with a variable-delay interferometer.

[0057] FIG. 10 shows a block diagram of yet another exemplary embodiment of the system according to the present invention. This exemplary system includes an interferometer, a probe, and a detection spectrometer which have been described in detail in elsewhere in detail S. H. Yun et al., “High-speed spectral domain optical coherence tomography at 1.3 μm wavelength,” Opt. Express 11, 3598-3604 (2003). In summary, this exemplary system included a light source 500, scanner driver 502, scanner clock generator 504, optical trigger generator 510 comprising a 5% tap 512, an (optional) optical narrowband filter 514, photodetector 516, and a TTL generating circuit 518. The narrowband filter 514 is used for a swept source operation, but is not needed for the pulsed broadband source operation. The interferometer can include a circulator 520, polarization controllers 522, polarizer 524, 10/90 coupler 534, collimator 536, neutral density filter 538, reference mirror 540, galvanometer-mounted mirror 542, galvanometer driver 544, imaging lens 546, sample 550. The detection arm may include a spectrometer 560 that has a collimator 562, grating 564, imaging lens 566, CCD linear array 570, camera 572. A galvanometer can be used in the probe to provide transverse beam scanning across a sample with a FWHM beam diameter and confocal length of 18 μm and 1.1 mm, respectively. The detection spectrometer 560, shown in the dash-dot box, consisted of a ruled diffraction grating 564 with 1,200 lines per mm, focusing lens 566 (f=150 mm), and a line scan camera (LSC) with a 512-element InGaAs CCD array (Sensors Unlimited Inc., SU512LX). Polarization controllers were adjusted to maximize the fringe visibility in the CCD. A total wavelength span of 106 nm centered at 1320 nm was projected to the 512-element CCD array with a spectral resolution of 0.1 nm.

[0058] The camera readout can be triggered by an external TTL signal generated from the source output. In the case of the pulsed light source, the electrical trigger pulses were generated directly from the optical pulses, as illustrated in the dotted box in FIG. 10. In the swept source case, the laser output may be transmitted through a combination of a circulator and a fiber Bragg grating reflector with 0.2 nm bandwidth and 90% reflectivity (the narrowband filter arrangement being presented by a small dotted box 510). The photodetector can then detect a train of short pulses generated when the output spectrum of the laser swept through the reflection band of the Bragg grating. From the photodetector output, TTL trigger pulses were generated with adjustable phase delay.

[0059] As described above, both lasers may be operated at a repetition rate of 18.939 kHz. This rate corresponded to the maximum readout rate of the camera. Upon receiving the trigger, the camera integrates photo-generated electrons for 24.4 μs; in the subsequent 28.4 μs period, the integrated voltage can be read out. By adjusting the phase delay in a PPL pulse generator, the integration time window of the camera was aligned to the output of the light sources, as shown in FIGS. 7(a) and (c). The camera output can be digitized with a 4-ch, 12-bit data acquisition board 582 (National Instruments, NI PCI-6115) and processed in a personal computer 584. The data processing may involve zero padding, interpolation and mapping to linear k-space, prior to a fast Fourier transform to create an image.

[0060] SD-OCT imaging can be performed using three different light sources: (a) the cw ASE obtained directly from the SOA, (b) the intensity-gated ASE pulses (as shown in FIG. 8(a)), and (c) the wavelength swept laser (as shown in FIG. 8(b)). In order to investigate motion artifacts, a sample can be constructed by mounting paper on an acoustic speaker. FIG. 11 shows exemplary images obtained with three different sources for comparison purposes. Shown on the left portion of FIG. 11 are exemplary OCT images acquired using cw, pulsed, and swept light, respectively, when the paper sample was kept stationary. Each image includes 256 axial and 500 transverse pixels, spans a depth of 2.1 mm and a width of 5 mm, and was acquired over a total time period of 26.4 ms. The images are generated using a logarithmic inverse grayscale over a dynamic range of 40 dB in reflectivity (as shown as a grayscale map in FIG. 11). For each of the light sources, the optical power illuminating the sample was adjusted approximately to the same level by using neutral density filters in the probe. The offset of the grayscale map for each light source can be finely adjusted so that the three static images (See FIG. 11, sections a, c, and e) exhibited nearly the same contrast. Images of the axially moving sample (see FIG. 11, sections b, d, and f) may be acquired when the speaker was driven with a sinusoidal waveform at 80 Hz with peak-to-peak amplitude of 0.8 mm. Signal fading due to fringe washout is distinct for the case of the cw ASE source (see FIG. 11, section b). Except near the peaks and valleys of the oscillation when the axial velocity is zero, the image contrast and penetration depth may be noticeably degraded. In contrast, the image d can be obtained with the pulsed source and exhibits considerably reduced image fading. Signal fading may not be observed while using the wavelength swept source (see FIG. 11, section f).

[0061] To quantify the amount of signal fading, a sum of the pixel values in the unit of linear power along each A-line may be obtained from the exemplary images shown in FIG. 11, representing a total signal power in the particular A-line. A total of 200 pixels, from the 31^stto 230^thelements, were considered in the summation. The results thereof are shown in FIGS. 12(a)-(c), such that the results in FIG. 12(a) corresponds to the signals obtained using to the cw source (see FIG. 11, sections a and b), the results in FIG. 12(b) corresponds to the signals obtained using the pulsed source (see FIG. 11, sections c and d), and the results in FIG. 12(c) corresponds to the signals obtained using the swept source (see FIG. 11, section e and f).

[0062] In each graph, the integrated signal power is plotted as a function of A-line index for the stationary-sample image (a lighter line) and the moving-sample image (a darker line). As depicted by the lighter lines, the signal power for the stationary sample exhibits random fluctuation due to speckle as the probe beam is scanned across the sample with standard deviation of approximately 2 dB. The speckle-averaged mean value varies linearly over transverse locations of the sample, a variation that was attributed to the finite confocal parameter and resulting depth-dependent light collection efficiency. The signal power traces obtained from FIG. 11, section b, d, and f (darker lines) clearly demonstrate the benefit of the pulsed and swept source in terms of reducing motion-induced signal fading.

[0063] The time gated pulses may provide a factor 8.6 reduction in signal integration time, from 24.4 μs to 2.85 μs. For the swept source with an instantaneous linewidth of 0.4 nm, individual CCD pixels may be illuminated for only 75 ns per each A-line acquisition representing a 325-fold reduction in signal integration time. Theoretical curves based on Eq. (1) show good correspondence with the experimental results with the following exceptions. The experimental noise floor can prohibit detection of signal loss greater than −14 dB; the small discrepancy between the blue and black curves in FIG. 12(c), by up to 3 dB, is attributed to the uneven probe collection efficiency at different depths of the two samples.

[0064] An exemplary SNR analysis indicates that the pulsed ASE source produced essentially the same noise characteristics as cw ASE of the same average optical power. However, images which may be acquired using the wavelength swept laser exhibited a noise floor that can be 10-20 dB higher, depending on depth, than that observed when using the ASE source of the same average power. We attribute this increased noise floor to the RIN of the swept laser in the frequency band from DC to 41 kHz corresponding to a reciprocal of the CCD integration time. The best sensitivity obtained with the swept source may be approximately −95 dB at a reference-arm power of 1-2 μW.

[0065] Exemplary SD-OCT imaging of a human coronary artery in vitro may be conducted by use of a fiber-optic catheter. The fiber-optic catheter comprised a graded-index lens and a 90-degree prism at its distal end and was connected to the interferometer through a high-speed rotational joint which could provide a rotational speed of up 100 revolutions per second (rps). FIG. 13 shows exemplary images obtained with the cw ASE source (see images A and B of FIG. 13) and the swept source (see images C and D of FIG. 13) at the same A-line acquisition rate of 18.94 kHz. The difference between images of images A and B and images C and D is the rotational speed of the catheter, which was 9.5 rps for images A and C, corresponding to 2000 A-lines per image, and 37.9 rps for images B and D, corresponding to 500 A-lines per image. Zero delay of the interferometer was positioned between the sample and the outer prism surface, resulting in a circular artifact superimposed on the image of the tissue (marked asp).

[0066] Image A may represent a typical OCT image of a vessel. In contrast, Image B can exhibit distinct radial streaks due to loss of signal. This image fading may be attributed mainly to catheter-induced modulation in path length, increasing with the rotational speed. The path length modulation can result from three mechanisms: (a) rotational beam scanning of an off-center object inevitably results in axial path length variation of the probe beam, as if the probe was retracting or approaching to the sample; (b) the tip of a rotating catheter can wobble in a protection sheath to modulate the distance between the probe and the sample; (c) mechanical vibration from a rotation joint can modulate the length of the optical fiber inside the catheter by twist or strain. Such third mechanism was thought to a dominant cause in this particular experiment, since the circle (p) corresponding to the prism surface also suffers from significant loss of contrast at the same radial locations. Images C and D of FIG. 13 are exemplary SD-OCT images obtained with the swept source. The signal fading is not noticeable in image D, demonstrating clearly the benefit of the pulsed-source approach.

[0067] Thus, multiple strategies can be applied to realize the benefit of pulsed or gated illumination. Traditional light sources include cw SLD's, supercontinuum sources, or mode-locked lasers. Each of these sources can be converted into a pulsed source by use of an external intensity modulation scheme. As an intensity modulator or switch, one may consider electro-optic or acousto-optic modulators or injection current modulation. Alternatively, CCD cameras with built-in electrical shutters may be used. This external gating approach, however, has a main drawback in that it results in a loss of optical power and therefore may degrade the detection sensitivity. However, in situations where motion causes significant signal fading through fringe washout, external gating can lead to a better sensitivity despite the loss of optical power. In other applications, however, the usable optical power in the system is often limited by the maximum permissible exposure of the sample. In this case, external gating would be an effective way to attenuate the power level entering the system from a powerful source. For example, ophthalmologic retinal imaging has been performed with SD-OCT at a wavelength of 800-nm. At this wavelength, the maximum permissible cw exposure to the eyes is limited to approximately 600-700 μW according to American National Standards Institute (ANSI). For this application, one could gate the output from a commercially available mode-locked Ti:Sapphire laser and, while still providing sufficient power to the system, reduce sensitivity to motion by more than an order of magnitude.

[0068] Instead of external gating, various power-efficient internal modulation techniques may be employed. For example, Q-switching and cavity dumping are well known techniques applicable to ultrashort pulsed lasers. Q-switched supercontinuum sources with repetition rates of a few to tens of kHz have been reported and may be suitable for use in the exemplary SD-OCT systems. Beside the benefit of reducing motion artifacts, the reduced fringe washout of the pulsed source approach may also facilitate quadrature fringe detection based on sequential phase dithering.

[0069] The use of a wavelength swept source as described in this manuscript is essentially a hybrid between the OFDI and SD-OCT techniques that may permit otherwise less-flexible OFDI source requirements including narrow instantaneous linewidth and tuning linearity to be relaxed. In this case, the high resolution and linearity of the spectrometer can accommodate a swept laser with a nonlinear tuning element such as a resonantly scanned Fabry-Perot filter or a tunable source based on soliton self-frequency shifting in nonlinear fibers. Furthermore, the relaxed requirement on the instantaneous linewidth of a swept laser may facilitate the generation of higher output powers.

[0070] In another exemplary embodiment of the system according to the present invention, each of the CCD arrays can be a 2-dimensional array. Two dimensional simultaneous scanning can be performed by using the 2-dimensional array, where along one axis of the array spectral information is encoded, while across the second dimension spatial information is encoded. FIG. 14 shows a block diagram of such exemplary system which may include a line source 600, lens 604, beam splitter 610, and a two-dimensional CCD array 620. The tissue is preferably illuminated by a line beam, and the illuminated portion in the sample is imaged on one dimension of the array, while the light is spectrally dispersed in the other direction of the array. As previously discussed, long integration times with a continuous source lead to motion artifacts and fringe washout. Also, read-out times of 2-dimensional arrays are larger that of 1-dimensional arrays. By using a pulsed source, motion artifacts and fringe washout can be avoided, where the exposure time of the array is significantly shorter than the frame transfer rate. Since the light intensity is distributed over a line, more power is allowed to be incident on the tissue. When using a pulsed source with pulse durations longer than 100 femtoseconds, the source can be treated as semi-continuous in ophthalmic applications. Thus, high peak power can be used over short periods of time, while the average power is in compliance with ANSI standards for light exposure of tissue. In addition, a swept source can be used in combination with line illumination, where the detector can be a 1 or 2 dimensional array. In case of a one dimensional array, the tissue information over a full line is acquired for each wavelength consecutively by a 1-dimensional array. By using a 2-dimensional array, the wavelength is encoded along the second dimension of the array.

[0071] As yet another exemplary embodiment of the present invention, a pulsed source can be employed in full-field optical coherence tomography, as depicted in FIG. 15 as a block diagram. The detector array is typically a 2-dimensional CCD array. The operating principle and generic system architecture for full-field OCT is well known in the art. Full-field OCT typically produces en face images. As with the earlier description of SD-OCT, the pulsed-source approach effectively reduces the effective signal acquisition time in a CCD array which is typically two dimensional. The repetition rate is matched to the frame readout rate of the CCD array. Since a typical full-field OCT technique is not based on spectral-domain interferometry, the swept source approach does not provide an advantage in full-field OCT in terms of motion artifacts. Nevertheless, a swept source whose sweep repetition rate is matched to the CCD readout rate may be still usable for full-field OCT as an alternative source to conventional broadband source. This is because of the fact that a swept spectrum seen by the CCD in a time-integrated manner is identical to a broadband spectrum of a same spectral envelope. The light source 700 is preferably spatially incoherent source, such as Halogen or Tungsten lamp in Kohler configuration, but operated in a pulsed regime to reduce the motion artifacts in full-field imaging. The source beam is divided into a reference and a sample by a beam splitter. High-NA objective lenses 710 are typically used. The reference mirror 540 may be attached to a mechanical actuator such as PZT for phase dithering to realize heterodyne detection.

[0072] The invention disclosed here may be used in various imaging applications, ranging from coronary artery imaging, GI tract, ophthalmologic imaging, to monitoring of dynamic biological or chemical process, moving materials and components, where high-sensitivity, high-speed, motion-artifact-free imaging is preferred.

[0073] 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. For example, the invention described herein is usable with the exemplary methods, systems and apparatus described in U.S. Provisional Patent Application No. 60/514,769 filed Oct. 27, 2003, and International Patent Application No. PCT/US03/02349 filed on Jan. 24, 2003, the disclosures of which are incorporated by reference 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, all publications, patents and patent applications referenced above are incorporated herein by reference in their entireties.

Claims

What is claimed is:

1. A system for imaging at least a portion of a sample, comprising:

a source arrangement generating at least one source electro-magnetic radiation forwarded to the sample and a reference; and

at least one detection arrangement including a plurality of detectors, at least one of the detectors capable of detecting a signal associated with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference,

wherein at least one particular detector of the detectors has a particular electrical integration time, wherein the at least one particular detector receives at least a portion of the signal for a time duration which has at least one first portion with at least one first power level greater than a predetermined threshold and at least one second portion immediately preceding or following the at least one first portion, the at least one second portion having at least one second power level less than the predetermined threshold, and wherein the at least one second portion is extended for a time period which is approximately at least 10% of the particular electrical integration time.

2. The system according to claim 1, wherein the signal has frequency components of the combination.

3. The system according to claim 1, wherein the source arrangement is a pulsed broadband source.

4. The system according to claim 1, wherein the detection arrangement includes at least one charged-coupled device.

5. The system according to claim 2, wherein the at least one particular detector receives the at least one of the frequency components.

6. The system according to claim 5, wherein the source arrangement is a pulsed broadband source.

7. The system according to claim 5, wherein the source arrangement includes an optical gating switch.

8. The system according to claim 5, wherein a frequency of the at least one source electro-magnetic radiation varies over time.

9. The system according to claim 5, wherein the detector arrangement further includes an electrical shutter that is adapted to gate a transmission of photoelectrons associated with the combination of the first and second electro-magnetic radiation, wherein a time period for the gating to allow the transmission of the photoelectrons is less than approximately 90% of the particular electrical integration time.

10. The system according to claim 5, wherein the sample is a biological sample.

11. The system according to claim 5, wherein the detection arrangement includes at least one charged-coupled device.

12. The system according to claim 5, further comprising at least one spectral separating unit which separates spectrum of at least one of the first electro-magnetic radiation, the second electro-magnetic radiation and the combination into the at least one of the frequency components.

13. The system according to claim 6, wherein the at least one source electro-magnetic radiation generated by the pulsed source is a single pulse per the particular electrical integration time.

14. The system according to claim 6, wherein the at least one source electro-magnetic radiation generated by the source arrangement is a burst of radiation that extends for at most approximately 90% of the particular electrical integration time.

15. The system according to claim 6, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a spectrum with a center wavelength between approximately 700 nanometers and 2000 nanometers.

16. The system according to claim 6, wherein a duration of the burst of radiation is approximately shorter than 1 μsec.

17. The system according to claim 8, wherein a mean frequency of the at least one source electro-magnetic radiation changes substantially continuously over time at a tuning speed that is greater than 100 terahertz per millisecond.

18. The system according to claim 8, wherein the mean frequency changes repeatedly with a repetition period that is less than approximately 90% of the particular electrical integration time.

19. The system according to claim 8, wherein the at least one source electro-magnetic radiation generated by the source arrangement has a tuning range with a center wavelength between approximately 700 nanometers and 2000 nanometers.

20. The system according to claim 8, wherein the at least one source electro-magnetic radiation generated by the source arrangement has a tuning range of approximately greater than 1% of the center wavelength.

21. The system according to claim 8, wherein the at least one source electro-magnetic radiation generated by the source arrangement has an instantaneous line width and a tuning range, the instantaneous line width being less than approximately 10% of the tuning range.

22. The system according to claim 8, wherein the source arrangement includes a tunable laser.

23. The system according to claim 8, wherein the source arrangement includes a tunable filter.

24. The system according to claim 8, wherein the source arrangement includes a medium, and wherein the source arrangement generates the at least one source electro-magnetic radiation based on a non-linearity associated with the medium.

25. The system according to claim 8, wherein the frequency varies substantially linearly with time.

26. The system according to claim 8, wherein the frequency varies substantially sinusoidally with time.

27. The system according to claim 13, wherein the pulsed source includes at least one of a Q-switched laser, a cavity-dumped mode-lock laser, and a gain-switched laser.

28. The system according to claim 13, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a pulse width approximately shorter than 1 μsec.

29. The system according to claim 14, wherein the burst of radiation includes multiple pulses.

30. The system according to claim 15, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a spectrum with a spectral width of approximately greater than 1% of the center wavelength.

31. A method for imaging at least a portion of a sample, comprising:

generating at least one source electro-magnetic radiation forwarded to the sample and a reference; and

detecting at least a portion of a signal associated with a combination of at least one first electro-magnetic radiation received from the sample and at least one second electro-magnetic radiation received from the reference using at least one detector of a plurality of detectors of a detection arrangement,

wherein at least one particular detector of the detectors has a particular electrical integration time, and wherein the at least one particular detector receives at least a portion of the signal for a time duration which has a first portion with a first power level greater than a predetermined threshold and a second portion immediately preceding or following the first portion, the second portion having a second power level less than the predetermined threshold, and extending for a time period which is approximately more than 10% of the particular electrical integration time.

32. The method according to claim 31, wherein the signal has frequency components of the combination.

33. The method according to claim 31, wherein the generating step is performed by a source arrangement which is a pulsed broadband source.

34. The method according to claim 31, wherein the detection step is performed by a detection arrangement which includes at least one charged-coupled device.

35. The method according to claim 32, wherein the at least one particular detector receives the at least one of the frequency components.

36. The method according to claim 35, wherein the generating step is performed by a source arrangement which is a pulsed broadband source.

37. The method according to claim 35, wherein the source arrangement includes an optical gating switch.

38. The method according to claim 35, wherein a frequency of the at least one source electro-magnetic radiation varies over time.

39. The method according to claim 35, wherein the detector arrangement further includes an electrical shutter that is adapted to gate a transmission of photoelectrons associated with the combination of the first and second electro-magnetic radiation, wherein a time period for the gating to allow the transmission of the photoelectrons is less than approximately 90% of the particular electrical integration time.

40. The method according to claim 35, wherein the sample is a biological sample.

41. The method according to claim 35, wherein the detecting step is performed by a detection arrangement which includes at least one charged-coupled device.

42. The method according to claim 35, further comprising the step of separating spectrum of at least one of the first electro-magnetic radiation, the second electro-magnetic radiation and the combination into the at least one of the frequency components.

43. The method according to claim 36, wherein the at least one source electro-magnetic radiation generated by the pulsed source is a single pulse per the particular electrical integration time.

44. The method according to claim 36, wherein the at least one source electro-magnetic radiation generated by the source arrangement is a burst of radiation that extends for at most approximately 90% of the particular electrical integration time.

45. The method according to claim 36, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a spectrum with a center wavelength between approximately 700 nanometers and 2000 nanometers.

46. The method according to claim 36, wherein a duration of the burst of radiation is approximately shorter than 1 μsec.

47. The method according to claim 38, wherein a mean frequency of the at least one source electro-magnetic radiation changes substantially continuously over time at a tuning speed that is greater than 100 terahertz per millisecond.

48. The method according to claim 38, wherein the mean frequency changes repeatedly with a repetition period that is less than approximately 90% of the particular electrical integration time.

49. The method according to claim 38, wherein the at least one source electro-magnetic radiation generated by the source arrangement has a tuning range with a center wavelength between approximately 700 nanometers and 2000 nanometers.

50. The method according to claim 38, wherein the at least one source electro-magnetic radiation generated by the source arrangement has a tuning range of approximately greater than 1% of the center wavelength.

51. The method according to claim 38, wherein the at least one source electro-magnetic radiation generated by the source arrangement has an instantaneous line width and a tuning range, the instantaneous line width being less than approximately 10% of the tuning range.

52. The method according to claim 38, wherein the source arrangement includes a tunable laser.

53. The method according to claim 38, wherein the source arrangement includes a tunable filter.

54. The method according to claim 38, wherein the source arrangement includes a medium, and wherein the source arrangement generates the at least one source electro-magnetic radiation based on a non-linearity associated with the medium.

55. The method according to claim 38, wherein the frequency varies substantially linearly with time.

56. The method according to claim 38, wherein the frequency varies substantially sinusoidally with time.

57. The method according to claim 43, wherein the pulsed source includes at least one of a Q-switched laser, a cavity-dumped mode-lock laser, and a gain-switched laser.

58. The method according to claim 43, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a pulse width approximately shorter than 1 μsec.

59. The method according to claim 44, wherein the burst of radiation includes multiple pulses.

60. The method according to claim 45, wherein the at least one source electro-magnetic radiation generated by the pulsed broadband source has a spectrum width a spectral width of approximately greater than 1% of the center wavelength.

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Abstract