|Numéro de publication||USRE42497 E1|
|Type de publication||Octroi|
|Numéro de demande||US 12/205,248|
|Date de publication||28 juin 2011|
|Date de dépôt||5 sept. 2008|
|Date de priorité||6 mai 2003|
|État de paiement des frais||Payé|
|Autre référence de publication||US7102758, US20040223162, WO2004102109A2, WO2004102109A3|
|Numéro de publication||12205248, 205248, US RE42497 E1, US RE42497E1, US-E1-RE42497, USRE42497 E1, USRE42497E1|
|Cessionnaire d'origine||Duke University|
|Exporter la citation||BiBTeX, EndNote, RefMan|
|Citations de brevets (72), Citations hors brevets (37), Référencé par (7), Classifications (5), Événements juridiques (2)|
|Liens externes: USPTO, Cession USPTO, Espacenet|
1. Field of the Invention
The present invention relates to an apparatus and method for obtaining depth-resolved spectra for the purpose of determining structure by measuring elastic scattering properties. More particularly, Fourier domain, low-coherence interferometry techniques are applied to light scattering spectroscopy. This approach permits the viewing and recovery of depth-resolved structures, as well as obtaining spectroscopic information about scattered light as a function of depth.
2. Background of the Related Art
Accurately measuring small objects or other physical phenomena is a goal that is pursued in many diverse fields of scientific endeavor. For example, in the study of cellular biology and cellular structures, light scattering spectroscopy (LSS) has received much attention recently as a means for probing cellular morphology and the diagnosing of dysplasia. The disclosures of the following references are incorporated by reference in their entirety:
The LSS technique examines variations in the elastic scattering properties of cell organelles to infer their sizes and other dimensional information. In order to measure cellular features in tissues and other cellular structures, it is necessary to distinguish the singly scattered light from diffuse light, which has been multiply scattered and no longer carries easily accessible information about the scattering objects. This distinction or differentiation can be accomplished in several ways, such as the application of a polarization grating, by restricting or limiting studies and analysis to weakly scattering samples, or by using modeling to remove the diffuse component (s).
As an alternative approach for selectively detecting singly scattered light from sub-surface sites, low-coherence interferometry (LCI) has also been explored as a method of LSS. Experimental results have shown that using a broadband light source and its second harmonic allows the recovery of information about elastic scattering using LCI .
More recently, angle-resolved LCI (a/LCI) has demonstrated the capability of obtaining structural information by examining the angular distribution of scattered light from the sample or object under examination. The a/LCI technique has been successfully applied to measuring cellular morphology and to diagnosing intraepithelial neoplasia in an animal model of carcinogenesis.
The above references are incorporated by reference herein where appropriate for appropriate teachings of additional or alternative details, features and/or technical background.
The claimed exemplary embodiments of the present invention address some of the issues presented above.
An object of the invention is to solve at least the above problems and/or disadvantages and to provide at least the advantages described hereinafter.
In one exemplary embodiment of the present invention, an apparatus comprises a first receiver that receives a first reference light and outputs a second reference light. A second receiver that receives a first sample light and outputs a second sample light and wherein the second sample light contains light scattered from a sample when at least a portion of the first sample light is scattered from a sample. A cross-correlator that receives and cross-correlates the second reference light with the second sample light. The cross-correlator may be a spatial cross-correlator.
In another exemplary embodiment of the present invention, a reference arm receives a first reference light and outputs a second reference light. A sample receives a first sample light and outputs a second sample light and wherein the second sample light contains light scattered from the sample when at least a portion of said first sample light is scattered from the sample. A spatial cross-correlator receives and cross correlates the second reference light with the second sample light. The spatial cross-correlator comprises a detector and a processing unit. The detector outputs an interference term to the processing unit. The processing unit processes the interference term to yield depth resolved cross-correlation reflection profiles of the sample. The processing unit first applies a Gaussian window and then a Fourier transform transforms the interference term to yield depth resolved cross-correlation reflection profiles of the sample. The Fourier transform obtains an axial spatial cross-correlation between a signal field(s) and a reference field(s). A light source outputs light, which contains the first sample light and the first reference light.
In another exemplary embodiment of the present invention, a method comprises receiving a first reference light and outputting a second reference light. A first sample light is received and a second sample light is output. The second sample light contains light scattered from a sample when at least a portion of the first sample light is scattered from a sample along with the reception and cross correlation of the second reference light with the second sample light.
In another exemplary embodiment, a method comprises receiving light and splitting at least a portion of the light into reference light and sample light. At least a portion of said reference light is reflected from a reference surface to yield reflected reference light. At least a portion of the sample light is scattered from a sample to yield scattered sample light, and the scattered sample and the reflected reference light are mixed. Information is recovered about the scattered sample light. The mixing comprises detecting an intensity of the scattered sample light and the reflected reference light. Recovering information comprises extracting an interference term from a total intensity. Recovering information can further comprise applying a mathematical operator to the interference term to recover the spectral information about the scattered sample light at a particular depth to yield depth resolved cross-correlation reflection points of the sample. The mathematical operator used is preferably a Gaussian window.
The invention will be described in detail with reference to the following drawings in which like reference numerals refer to like elements wherein:
In the following detailed description of the various exemplary embodiments, reference is made to the accompanying drawings that show, by way of illustration, specific embodiments in which the invention may be practiced. In the drawings, like numerals describe substantially similar components throughout the several views. These embodiments are described in sufficient detail to enable those skilled in the art to practice the invention. Other embodiments may be utilized without departing from the scope of the present invention. Moreover, it is to be understood that various embodiments of the invention, although different, are not necessarily mutually exclusive. For example, a particular feature, structure, or characteristic described in one embodiment may be included within other embodiments. Therefore, the following detailed description is not to be taken in a limiting sense. The scope of the present invention is delineated by the claims, along with the full scope of equivalents to which such claims are entitled.
The contents of the following references are incorporated by reference in their entirety: Wojtkowski, M., A. Kowalczyk, R. Leitgeb, and A. F. Fercher, Opt. Lett., 27(16): p. 1415-1417 (2002); Wojtkowski, M., R. Leitgeb, A. Kowalczyk, T. Bajraszewski, and A. F. Fercher, J. Biomed. Opt., 7(3): p. 457-463 (2002); Leitgeb, R., M. Wojtkowski, A. Kowalczyk, C. K. Hitzenberger, M. Sticker, and A. F. Fercher, Opt. Lett., 25(11): p. 820-822 (2000).
In general, spectral radar makes use of techniques where depth-resolved structural information is recovered by applying a Fourier transform to the spectrum of two mixed fields. In fLCI, the aforementioned approach used in spectral radar applications is extended to recover not only depth-resolved structure, but also to obtain spectroscopic information about scattered light as a function of depth. The capabilities of fLCI enable extracting the size of polystyrene beads in a sub-surface layer based on their light scattering spectrum. The apparatus and method according to exemplary embodiments of the invention can be applied to many different areas. One such area of application is to recover nuclear morphology of sub-surface cell layers.
One exemplary embodiment of the fLCI scheme is shown in
This illumination scheme achieves Kohler illumination in that the fiber acts as a field stop, resulting in the proper alignment of incident or illuminating light and thereby achieving critical illumination of the sample. In the fLCI system 10, the white light beam is split by the beamsplitter 106 (BS) into a reference beam 105 and an input beam 107 to the sample 108. The light scattered by the sample 108 is recombined at the BS 106 with light reflected by the reference mirror 114 (M).
The reference beam 105 in conjunction with the reference mirror 114 forms a portion of a reference arm that receives a first reference light and outputs a second reference light. The input beam 107 and the sample 108 form a portion of a sample arm that receives a first sample light and outputs a second sample light.
Those skilled in the art will appreciate that the light beam can be split into a plurality of reference beams and input beams (e.g. N reference beams and N input beams) without departing from the spirit and scope of the present invention. Further, the splitting of the beams may be accomplished with a beamsplitter or a fiber splitter in the case of an optical fiber implementation of and exemplary embodiment of the present invention.
In the exemplary embodiment of the present invention shown in
The detected signal is linearly related to the intensity as a function of wavelength I(λ), which can be related to the signal and reference fields (Es, Er) as:
<I(λ)>=<|Es(λ)|2>+<|Er(λ)|2>+2Re<Es(λ)E*r(λ)>cos φ (1)
where φ is the phase difference between the two fields and <. . .> denotes an ensemble average.
The interference term is extracted by measuring the intensity of the signal and reference beams independently and subtracting them from the total intensity.
The axial spatial cross-correlation function, ΓSR(z) between the sample and reference fields is obtained by resealing the wavelength spectrum into a wavenumber (k=2π/λ) spectrum then Fourier transforming:
ΓSR(z)=∫dkeikz<Es(k)E*r(k)>cos φ. (2)
This term is labeled as an axial spatial cross-correlation as it is related to the temporal or longitudinal coherence of the two fields.
Another exemplary embodiment of an fLCI scheme is shown in
The reference light in reference fiber 125, in conjunction with a lens 126 (preferably an aspheric lens) and the reference mirror 128, forms a portion of a reference arm that receives a first reference light and outputs a second reference light. Specifically, reference light in reference fiber 125 is directed to the reference mirror 128 by lens 126, and the reference light reflected by the reference mirror 128 (second reference light) is coupled back into the reference fiber 125 with lens 126. The sample light in sample fiber 127 and the sample 130 form a portion of a sample arm that receives a first sample light and outputs a second sample light. Specifically, sample light in sample fiber 127 is directed to the sample 130 by lens 131 (preferably as aspheric lens), and at least a portion of the sample light scattered by the sample 130 is coupled into the sample fiber 127 by lens 131. In the exemplary embodiment shown in
At least a portion of the reflected reference light in reference fiber 125 and at least a portion of the scattered sample light on sample fiber 127 are coupled into a detector fiber 133 by the FS 124.
The output of detector fiber 133 coincides with the input of a miniature spectrograph 132, where the light is spectrally dispersed and detected.
From this autocorrelation, the coherence length of the field, lc=1.21 μm is determined. This is slightly larger than the calculated width of lc=2/Δkl/c=0.98 μm, with any discrepancy most likely attributed to uncompensated dispersion effects. Note that rescaling the field into wavenumber space is a nonlinear process which can skew the spectrum if not properly executed .
In data processing, a fitting algorithm is applied (e.g. a cubic spline fit) to the rescaled wavenumber spectrum and then resampled (e.g. resample with even spacing). The resampled spectrum is then Fourier transformed to yield the spatial correlation of the sample. Those skilled in the art will appreciate that other frequency based algorithms or combinations of algorithms can be used in place of the Fourier transform to yield spatial correlation. One example of a software tool that can be used to accomplish this processing in real time or near real time is to use LabView™ software.
In one exemplary embodiment of the present invention, the sample consists of a glass coverslip (e.g., thickness, d˜200 μm) with polystyrene beads which have been dried from suspension onto the back surface (1.55 μm mean diameter, 3% variance). Thus, the field scattered by the sample can be expressed as:
In equation 3, Efront and Eback denote the field scattered by the front and back surfaces of the coverslip, and δz is the difference between the path length of the reference beam and that of the light reflected from the front surface and n the index of refraction of the glass. The effect of the microspheres will appear in the Eback term as the beads are small and attached closely to the back surface. Upon substituting equation 3 into equation 2, a two peak distribution with the width of the peaks given by the coherence length of the source is obtained.
In order to obtain spectroscopic information, a Gaussian window is applied to the interference term before performing the Fourier transform operation. Those skilled in the art will appreciate that other probabilistic windowing methodologies may be applied without departing from the spirit and scope of the invention. This makes it possible to recover spectral information about light scattered at a particular depth.
The windowed interference term takes the form:
<Es(k)E*r(k)>exp [−((k−kw)/Δkw)2]. (4)
The proper sizing of a windowed interference term can facilitate the processing operation. For example, by selecting a relatively narrow window (Δkw small) compared to the features of Es and Ek, we effectively obtain <Es(kw)E*r(kw) >. In processing the data below, we use Δkw=0.12 μm−1 which degrades the coherence length by a factor of 16.7. This exemplary window setting enables the scattering at 50 different wavenumbers over the 6 μm−1 span of usable spectrum.
Note that the correlation function is symmetric about z=0, resulting in a superposed mirror image of the scan. Since these are represented as cross-correlation functions, the plots are symmetric about z=0. Thus the front surface reflection for z>0 is paired with the back surface reflection for z<0, and vice versa.
To obtain the spectrum of the scattered light, we repeatedly apply the Gaussian window and increase the center wavenumber by 0.12 μm−1 between successive applications. As mentioned above, Δkw=0.12 μm−1 is used to degrade the coherence length by a factor of 16.7. This results in the generation of a spectroscopic depth-resolved reflection profile.
For comparison, the same ratio for the front surface reflections (dashed line in
There are many applications of the various exemplary embodiments of the present invention. One exemplary application of fLCI is in determining the size of cell organelles, in particular the cell nucleus, in epithelial tissues. In biological media, for example, the relative refractive indices are lower for organelles compared to microspheres and thus, smaller scattering signals are expected. The use of a higher power light source will permit the smaller signals to be detected. Other examples include detection of sub-surface defects in manufactured parts, including fabricated integrated circuits, detection of airborne aerosols, such as nerve agents or biotoxins, and detection of exposure to such aerosols by examining epithelial tissues within the respiratory tract.
Additionally, the larger the size of the nucleus (compared to the microspheres in this experiment), the higher the frequency modulation of the spectrum. Those skilled in the art will appreciate that higher frequency oscillations are detected at a lower efficiency in Fourier transform spectroscopy techniques. Therefore, in order to detect these higher frequency oscillations, a higher resolution spectrograph is used.
The foregoing example illustrates how the exemplary embodiments of the present invention can be modified in various manners to improve performance in accordance with the spirit and scope of the present invention.
From the foregoing detailed description, it should be apparent that fLCI can recover structural information with sub-wavelength accuracy from sub-surface layers based on measuring elastic scattering properties. The simplicity of the system makes it an excellent candidate for probing cellular morphology in tissue samples and may one day serve as the basis for a biomedical diagnostic device.
The foregoing embodiments and advantages are merely exemplary and are not to be construed as limiting the present invention. The present teaching can be readily applied to other types of apparatuses. The description of the present invention is intended to be illustrative, and not to limit the scope of the claims. Many alternatives, modifications, and variations will be apparent to those skilled in the art. In the claims, means-plus-function clauses are intended to cover the structures described herein as performing the recited function and not only structural equivalents but also equivalent structures.
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|Brevet citant||Date de dépôt||Date de publication||Déposant||Titre|
|US8233152 *||9 juil. 2009||31 juil. 2012||Canon Kabushiki Kaisha||Optical coherence tomographic imaging method and optical coherence tomographic imaging apparatus|
|US8836952||19 juin 2012||16 sept. 2014||Canon Kabushiki Kaisha||Optical coherence tomographic imaging method and optical coherence tomographic imaging apparatus|
|US9274001 *||24 janv. 2011||1 mars 2016||Duke University||Dual window processing schemes for spectroscopic optical coherence tomography (OCT) and fourier domain low coherence interferometry|
|US9689745 *||22 févr. 2016||27 juin 2017||Duke University||Multiple window processing schemes for spectroscopic optical coherence tomography (OCT) and fourier domain low coherence interferometry|
|US20100027019 *||9 juil. 2009||4 févr. 2010||Canon Kabushiki Kaisha||Optical Coherence Tomographic Imaging Method and Optical Coherence Tomographic Imaging Apparatus|
|US20130135614 *||24 janv. 2011||30 mai 2013||Adam Wax||Dual window processing schemes for spectroscopic optical coherence tomography (oct) and fourier domain low coherence interferometry|
|WO2014144307A2||14 mars 2014||18 sept. 2014||Gebhart Steven C||Probe assembly and disposable cover particularly for use in endoscope applications of low coherence interferometry|
|Classification aux États-Unis||356/497|
|Classification internationale||G01B9/02, G01N15/02|
|20 sept. 2011||CC||Certificate of correction|
|6 févr. 2014||FPAY||Fee payment|
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