U.S. patent application number 13/551348 was filed with the patent office on 2012-11-08 for systems and methods for endoscopic angle-resolved low coherence interferometry.
This patent application is currently assigned to DUKE UNIVERSITY. Invention is credited to John W. Pyhtila, Adam Wax.
Application Number | 20120281224 13/551348 |
Document ID | / |
Family ID | 37714242 |
Filed Date | 2012-11-08 |
United States Patent
Application |
20120281224 |
Kind Code |
A1 |
Wax; Adam ; et al. |
November 8, 2012 |
SYSTEMS AND METHODS FOR ENDOSCOPIC ANGLE-RESOLVED LOW COHERENCE
INTERFEROMETRY
Abstract
Fourier domain a/LCI (faLCI) system and method which enables in
vivo data acquisition at rapid rates using a single scan.
Angle-resolved and depth resolved spectra information is obtained
with one scan. The reference arm can remain fixed with respect to
the sample due to only one scan required. A reference signal and a
reflected sample signal are cross-correlated and dispersed at a
multitude of reflected angles off of the sample, thereby
representing reflections from a multitude of points on the sample
at the same time in parallel. Information about all depths of the
sample at each of the multitude of different points on the sample
can be obtained with one scan on the order of approximately 40
milliseconds. From the spatial, cross-correlated reference signal,
structural (size) information can also be obtained using techniques
that allow size information of scatterers to be obtained from
angle-resolved data.
Inventors: |
Wax; Adam; (Chapel Hill,
NC) ; Pyhtila; John W.; (Durham, NC) |
Assignee: |
DUKE UNIVERSITY
Durham
NC
|
Family ID: |
37714242 |
Appl. No.: |
13/551348 |
Filed: |
July 17, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13042672 |
Mar 8, 2011 |
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13551348 |
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12538309 |
Aug 10, 2009 |
7903254 |
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13042672 |
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11548648 |
Oct 11, 2006 |
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12538309 |
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60725603 |
Oct 11, 2005 |
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Current U.S.
Class: |
356/456 ;
356/450; 356/498 |
Current CPC
Class: |
G01N 2021/4709 20130101;
G01B 9/02044 20130101; A61B 5/0066 20130101; G01B 9/0209 20130101;
A61B 5/0075 20130101; G01N 21/31 20130101; G01B 9/02084 20130101;
G01N 21/4795 20130101; A61B 5/0084 20130101; G01N 2021/4704
20130101; G01N 2021/4735 20130101; G01J 3/4531 20130101; G01J
3/4412 20130101; A61B 5/7257 20130101; G01N 2201/08 20130101; G01B
9/02087 20130101 |
Class at
Publication: |
356/456 ;
356/450; 356/498 |
International
Class: |
G01J 3/45 20060101
G01J003/45; G01B 11/22 20060101 G01B011/22; G01B 9/02 20060101
G01B009/02 |
Goverment Interests
STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0003] This invention was supported by the National Institute of
Health, Grant No. R21-CA-109907, and the National Science
Foundation, Grant No. BES-03-48204. The United States Government
has certain rights in the invention.
Claims
1. An apparatus comprising: a first arrangement configured to
receive at least one first electro-magnetic radiation, and forward
at least one second electro-magnetic radiation within a solid angle
to a sample, wherein the at least one second electro-magnetic
radiation is associated with the at least one first
electro-magnetic radiation, wherein the first arrangement is
configured to receive a plurality of third electro-magnetic
radiations from the sample which is associated with the at least
one second electro-magnetic radiation, and wherein at least one
portion of the third electro-magnetic radiations is provided
outside a periphery of the solid angle; and a second arrangement
configured to simultaneously detect signals which are (i) provided
along optical axes associated therewith that are different from one
another, and (ii) associated with each of the third
electro-magnetic radiations, wherein the signals are associated
with information for the at least one sample at a plurality of
depths thereof, and wherein the second arrangement is configured to
determine the depths using the at least one portion of the third
electro-magnetic radiations.
2. The apparatus according to claim 1, further comprising a third
arrangement configured to detect an interference between the at
least one portion of the third electro-magnetic radiation and at
least one fourth electro-magnetic radiation associated with the at
least one first electro-magnetic radiation, and to obtain
information associated with the sample as a function of the depths
within the sample based on the interference.
3. The apparatus according to claim 1, further comprising a third
arrangement configured to provide data associated with at least one
of spectroscopic properties, angular back-scattering properties or
elastic properties of at least one portion of the sample as a
function of the signals.
4. The apparatus according to claim 1, further comprising a third
arrangement configured to provide data associated with scattering
characteristics of at least one portion of the sample as a function
of a combination of the signals.
5. The apparatus according to claim 1, wherein the second
arrangement is configured to determine the depths using a single
one of the third electro-magnetic radiations.
6. The apparatus according to claim 1, wherein the second
arrangement is further configured to combine the signals.
7. A method for detecting signals, comprising: receiving at least
one first electro-magnetic radiation; forwarding at least one
second electro-magnetic radiation within a solid angle to a sample,
wherein the at least one second electro-magnetic radiation is
associated with the at least one first electro-magnetic radiation;
receiving a plurality of third electro-magnetic radiations from the
sample which is associated with the at least one second
electro-magnetic radiation, wherein at least one portion of the
third electro-magnetic radiations is provided outside a periphery
of the solid angle; simultaneously detecting the signals which are
(i) provided along optical axes associated therewith that are
different from one another, and (ii) associated with each of the
third electro-magnetic radiations, wherein the signals are
associated with information for the at least one sample at a
plurality of depths thereof, and using a computer arrangement,
determining the depths using the at least one portion of the third
electro-magnetic radiations.
8. The method according to claim 7, further comprising, after the
simultaneous detection, combining the signals.
9. An apparatus comprising: a first arrangement configured to
receive at least one first electro-magnetic radiation, and forward
at least one second electro-magnetic radiation within a solid angle
to a sample, wherein the at least one second electro-magnetic
radiation is associated with the at least one first
electro-magnetic radiation, wherein the first arrangement is
configured to simultaneously receive at least two of a plurality of
third electro-magnetic radiations from the sample which is
associated with the at least one second electro-magnetic radiation,
and wherein at least one portion of the third electro-magnetic
radiations is provided outside a periphery of the solid angle; and
a second arrangement configured to simultaneously detect an
interference between the at least two of the third radiations which
are provided along optical axes associated therewith that are
different from one another and at least one fourth radiation
associated with the at least one first radiation, and configured to
obtain information associated with the sample as a function of at
least one depth within the sample based on the interference.
10. The apparatus according to claim 9, wherein the second
arrangement is configured to determine the at least one depth based
on the interference.
11. The apparatus according to claim 9, wherein the one second
arrangement is configured to simultaneously detect signals
associated with each of the third electro-magnetic radiations.
12. The apparatus according to claim 11, further comprising a third
arrangement configured to provide data associated with at least one
of spectroscopic properties, angular back-scattering properties or
elastic properties of at least one portion of the sample as a
function of the signals.
13. The apparatus according to claim 11, further comprising a third
arrangement configured to provide data associated with scattering
characteristics of at least one portion of the sample as a function
of a combination of the signals.
14. The apparatus according to claim 10, wherein the second
arrangement is configured to determine the depths using a single
one of the third electro-magnetic radiations.
15. A method for detecting signals, comprising: receiving at least
one first electro-magnetic radiation; forwarding at least one
second electro-magnetic radiation within a solid angle to a sample,
wherein the at least one second electro-magnetic radiation is
associated with the at least one first electro-magnetic radiation;
simultaneously receiving at least two of a plurality of third
electro-magnetic radiations from the sample which is associated
with the at least one second electro-magnetic radiation, wherein at
least one portion of the third electro-magnetic radiations is
provided outside a periphery of the solid angle; simultaneously
detecting an interference between the at least two of the third
radiations and at least one fourth radiation associated with the at
least one first radiation wherein the third radiations are provided
along optical axes associated therewith that are different from one
another; and using a computer arrangement, obtaining information
associated with the sample as a function of at least one depth
within the sample based on the interference.
16. The method according to claim 15, further comprising
simultaneously detecting signals associated with each of the third
electro-magnetic radiations which are provided along optical axes
associated therewith that are different from one another.
Description
RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
patent application Ser. No. 13/042,672 entitled "SYSTEMS AND
METHODS FOR ENDOSCOPIC ANGLE-RESOLVED LOW COHERENCE INTERFEROMETRY"
filed Mar. 8, 2011, which is herein incorporated by reference in
its entirety and which is a continuation of U.S. patent application
Ser. No. 12/538,309, now U.S. Pat. No. 7,903,254 entitled "SYSTEMS
AND METHODS FOR ENDOSCOPIC ANGLE-RESOLVED LOW COHERENCE
INTERFEROMETRY," filed on Aug. 10, 2009, which is herein
incorporated by reference in its entirety and which is a
continuation application of U.S. patent application Ser. No.
11,548,648, now U.S. Pat. No. 7,595,889, entitled "SYSTEMS AND
METHODS FOR ENDOSCOPIC ANGLE-RESOLVED LOW COHERENCE
INTERFEROMETRY," filed on Oct. 11, 2006, which is herein
incorporated by reference in its entirety, which claims priority to
U.S. Provisional Patent Application No. 60/725,603 entitled
"SYSTEMS AND METHODS FOR ENDOSCOPIC ANGLE-RESOLVED LOW COHERENCE
INTERFEROMETRY," filed on Oct. 11, 2005, also incorporated herein
by reference in its entirety.
[0002] This application is also related to U.S. Pat. No. 7,102,758
entitled "FOURIER DOMAIN LOW-COHERENCE INTERFEROMETRY FOR LIGHT
SCATTERING SPECTROSCOPY APPARATUS AND METHOD," which is
incorporated herein by reference in its entirety.
FIELD
[0004] Fourier domain angle-resolved low coherence interferometry
(faLCI) system and method that enables data acquisition of
angle-resolved and depth-resolved spectra information of a sample,
in which depth and size information about the sample can be
obtained with a single scan at rapid rates for in vivo applications
in particular.
BACKGROUND
[0005] Examining the structural features of cells is essential for
many clinical and laboratory studies. The most common tool used in
the examination for the study of cells has been the microscope.
Although microscope examination has led to great advances in
understanding cells and their structure, it is inherently limited
by the artifacts of preparation. The characteristics of the cells
can only been seen at one moment in time with their structure
features altered because of the addition of chemicals. Further,
invasion is necessary to obtain the cell sample for
examination.
[0006] Thus, light scattering spectrography (LSS) was developed to
allow for in vivo examination applications, including cells. 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).
[0007] 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. LCI
utilizes a light source with low temporal coherence, such as
broadband white light source for example. Interference is only
achieved when the path length delays of the interferometer are
matched with the coherence time of the light source. The axial
resolution of the system is determined by the coherent length of
the light source and is typically in the micrometer range suitable
for the examination of tissue samples. Experimental results have
shown that using a broadband light source and its second harmonic
allows the recovery of information about elastic scattering using
LCI. LCI has used time depth scans by moving the sample with
respect to a reference arm directing the light source onto the
sample to receive scattering information from a particular point on
the sample. Thus, scan times were on the order of 5-30 minutes in
order to completely scan the sample.
[0008] Angle-resolved LCI (a/LCI) has been developed as a means to
obtain sub-surface structural information regarding the size of a
cell. Light is split into a reference and sample beam, wherein the
sample beam is projected onto the sample at different angles to
examine the angular distribution of scattered light. The a/LCI
technique combines the ability of (LCI) to detect singly scattered
light from sub-surface sites with the capability of light
scattering methods to obtain structural information with
sub-wavelength precision and accuracy to construct depth-resolved
tomographic images. Structural information is determined by
examining the angular distribution of the back-scattered light
using a single broadband light source is mixed with a reference
field with an angle of propagation. The size distribution of the
cell is determined by comparing the osciallary part of the measured
angular distributions to predictions of Mie theory. Such a system
is described in Cellular Organization and Substructure Measured
Using Angle-Resolved Low-Coherence Interferometry, Biophysical
Journal, 82, April 2002, 2256-2265, incorporated herein by
reference in its entirety.
[0009] The a/LCI technique has been successfully applied to
measuring cellular morphology and to diagnosing intraepithelial
neoplasia in an animal model of carcinogenesis. The inventors of
the present application described such a system in Determining
nuclear morphology using an improved angle-resolved low coherence
interferometry system in Optics Express, 2003, 11(25): p.
3473-3484, incorporated herein by reference in its entirety. The
a/LCI method of obtaining structural information about a sample has
been successfully applied to measuring cellular morphology in
tissues and in vitro as well as diagnosing intraepithelial
neoplasia and assessing the efficacy of chemopreventive agents in
an animal model of carcinogenesis. a/LCI has been used to
prospectively grade tissue samples without tissue processing,
demonstrating the potential of the technique as a biomedical
diagnostic.
[0010] Initial prototype and second generation a/LCI systems
required 30 and 5 minutes respectively to obtain similar data.
These earlier systems relied on time domain depth scans just as
provided in previous LCI based systems. The length of the reference
arm of the interferometer had to be mechanically adjusted to
achieve serial scanning of the detected scattering angle. The
method of obtaining angular specificity was achieved by causing the
reference beam of the interferometry scheme to cross the detector
plane at a variable angle. This general method for obtaining
angle-resolved, depth-resolved backscattering distributions was
disclosed in U.S. Pat. No. 6,847,456 entitled "Methods and systems
using field-based light scattering spectroscopy," which is
incorporated by reference herein in its entirety.
[0011] Other LCI prior systems are disclosed in U.S. Pat. Nos.
6,002,480 and 6,501,551, both of which are incorporated by
reference herein in their entireties. U.S. Pat. No. 6,002,480
covers obtaining depth-resolved spectroscopic distributions and
discusses obtaining the size of scatterers by observing changes in
wavelength due to elastic scattering properties. U.S. Pat. No.
6,501,551 covers endoscopic application of interferometric imaging
and does anticipate the use of Fourier domain concepts to obtain
depth resolution. U.S. Pat. No. 6,501,551 does not discuss
measurement of angularly resolved scattering distributions, the use
of scattered light to determine scatterer size by analysis of
elastic scattering properties, nor the use of an imaging
spectrometer to record data in parallel, whether that data is
scattering or imaging data. Finally, U.S. Pat. No. 7,061,622
discusses fiber optic means for measuring angular scattering
distributions, but does not discuss the Fourier domain concept.
Also because it describes an imaging technique, the embodiments all
include focusing optics which limit the region probed.
SUMMARY OF THE DETAILED DESCRIPTION
[0012] Embodiments disclosed herein involve a new a/LCI technique
called Fourier domain a/LCI (faLCI), which enables data acquisition
at rapid rates using a single scan, sufficient to make in vivo
applications feasible. The embodiments disclosed herein obtain
angle-resolved and depth-resolved spectra information about a
sample, in which depth and size information about the sample can be
obtained with a single scan, and wherein the reference arm can
remain fixed with respect to the sample due to only one scan
required. A reference signal and a reflected sample signal are
cross-correlated and dispersed at a multitude of reflected angles
off of the sample, thereby representing reflections from a
multitude of points on the sample at the same time in parallel.
[0013] Since this angle-resolved, cross-correlated signal is
spectrally dispersed, the new data acquisition scheme is
significant as it permits data to be obtained in less than one
second, a threshold determined to be necessary for acquiring data
from in vivo tissues. Information about all depths of the sample at
each of the multitude of different points on the sample can be
obtained with one scan on the order of approximately 40
milliseconds. From the spatial, cross-correlated reference signal,
structural (size) information can also be obtained using techniques
that allow size information of scatterers to be obtained from
angle-resolved data.
[0014] The faLCI technique of the disclosed embodiments uses the
Fourier domain concept to acquire depth resolved information.
Signal-to-noise and commensurate reductions in data acquisition
time are possible by recording the depth scan in the Fourier (or
spectral) domain. The faLCI system combines the Fourier domain
concept with the use of an imaging spectrograph to spectrally
record the angular distribution in parallel. Thereafter, the
depth-resolution of the disclosed embodiments is achieved by
Fourier transforming the spectrum of two mixed fields with the
angle-resolved measurements obtained by locating the entrance slit
of the imaging spectrograph in a Fourier transform plane to the
sample. This converts the spectral information into depth-resolved
information and the angular information into a transverse spatial
distribution. The capabilities of faLCI have been initially
demonstrated by extracting the size of polystyrene beads in a
depth-resolved measurement.
[0015] Various mathematical techniques and methods are provided for
determining size information of the sample using the
angle-resolved, cross-correlated signal.
[0016] The embodiments disclosed herein are not limited to any
particular arrangement. In one embodiment, the apparatus is based
on a modified Mach-Zehnder interferometer, wherein broadband light
from a superluminescent diode is split into a reference beam and an
input beam to the sample by a beamsplitter. In another embodiment,
a unique optical fiber probe can be used to deliver light and
collect the angular distribution of scattered light from the sample
of interest.
[0017] The a/LCI method can be a clinically viable method for
assessing tissue health without the need for tissue extraction via
biopsy or subsequent histopathological evaluation. The a/LCI system
can be applied for a number of purposes: early detection and
screening for dysplastic epithelial tissues, disease staging,
monitoring of therapeutic action and guiding the clinician to
biopsy sites. The non-invasive, non-ionizing nature of the optical
a/LCI probe means that it can be applied frequently without adverse
effect. The potential of a/LCI to provide rapid results will
greatly enhance its widespread applicability for disease
screening.
BRIEF DESCRIPTION OF THE FIGURES
[0018] The accompanying drawing figures incorporated in and forming
a part of this specification illustrate several aspects of the
disclosed embodiments, and together with the description serve to
explain the principles of the disclosed embodiments.
[0019] FIG. 1A is a schematic of one exemplary embodiment of the
faLCI system employing Mach-Zehnder interferometer;
[0020] FIG. 1B is an illustration showing the relationship of the
detected scattering angle to slit of spectrograph in the
interferometer arrangement of FIG. 1A;
[0021] FIG. 2 is a flowchart illustrating the steps performed by
the interferometer apparatus to recover depth-resolved spatial
cross-correlated information about the sample for analysis;
[0022] FIGS. 3A-D illustrate examples of faLCI data recovered in
the spectral domain for an exemplary sample of polystyrene beads,
comprising the total acquired signal (FIG. 3A), the reference field
intensity (FIG. 3B), the signal field intensity (FIG. 3C), and the
extracted, cross-correlated signal between the reference and signal
field intensities (FIG. 3D);
[0023] FIG. 4A is an illustration of the axial spatial
cross-correlated function performed on the cross-correlated faLCI
data illustrated in FIG. 3D as a function of depth and angle;
[0024] FIG. 4B is an illustration of an angular distribution plot
of raw and filtered data regarding scattered sample signal
intensity as a function of angle in order to recover size
information about the sample;
[0025] FIG. 5A is an illustration of the filtered angular
distribution of the scattered sample signal intensity compared to
the best fit Mie theory to determine size information about the
sample;
[0026] FIG. 5B is a Chi-squired minimization of size information
about the sample to estimate the diameter of cells in the
sample;
[0027] FIG. 6 is a schematic of exemplary embodiment of the faLCI
system employing an optical fiber probe;
[0028] FIG. 7A is a cutaway view of an a/LCI fiber-optic probe tip
that may be employed by the faLCI system illustrated in FIG. 6;
[0029] FIG. 7B illustrates the location of the fiber probe in the
faLCI system illustrated in FIG. 7A;
[0030] FIG. 8A is an illustration of an alternative fiber-optic
faLCI system that may be employed with the disclosed
embodiments;
[0031] FIG. 8B is an illustration of sample illumination and
scattered light collection with distal end of probe in the faLCI
system illustrated in FIG. 8B; and
[0032] FIG. 8C is an illustration of an image of the illuminated
distal end of probe of the faLCI system illustrated in FIG. 8A.
DETAILED DESCRIPTION
[0033] The embodiments set forth below represent the necessary
information to enable those skilled in the art to practice the
disclosed embodiments and illustrate the best mode of practicing
the embodiments. Upon reading the following description in light of
the accompanying drawing figures, those skilled in the art will
understand the concepts of the embodiments and will recognize
applications of these concepts not particularly addressed herein.
It should be understood that these concepts and applications fall
within the scope of the disclosure and the accompanying claims.
[0034] Embodiments disclosed herein involve a new a/LCI technique
called Fourier domain a/LCI (faLCI), which enables data acquisition
at rapid rates using a single scan, sufficient to make in vivo
applications feasible. The embodiments disclosed herein obtain
angle-resolved and depth-resolved spectra information about a
sample, in which depth and size information about the sample can be
obtained with a single scan, and wherein the reference arm can
remain fixed with respect to the sample due to only one scan
required. A reference signal and a reflected sample signal are
cross-correlated and dispersed at a multitude of reflected angles
off of the sample, thereby representing reflections from a
multitude of points on the sample at the same time in parallel.
[0035] Since this angle-resolved, cross-correlated signal is
spectrally dispersed, the new data acquisition scheme is
significant as it permits data to be obtained in less than one
second, a threshold determined to be necessary for acquiring data
from in vivo tissues. Information about all depths of the sample at
each of the multitude of different points on the sample can be
obtained with one scan on the order of approximately 40
milliseconds. From the spatial, cross-correlated reference signal,
structural (size) information can also be obtained using techniques
that allow size information of scatterers to be obtained from
angle-resolved data.
[0036] The faLCI technique of the disclosed embodiments uses the
Fourier domain concept to acquire depth resolved information.
Signal-to-noise and commensurate reductions in data acquisition
time are possible by recording the depth scan in the Fourier (or
spectral) domain. The faLCI system combines the Fourier domain
concept with the use of an imaging spectrograph to spectrally
record the angular distribution in parallel. Thereafter, the
depth-resolution of the disclosed embodiments is achieved by
Fourier transforming the spectrum of two mixed fields with the
angle-resolved measurements obtained by locating the entrance slit
of the imaging spectrograph in a Fourier transform plane to the
sample. This converts the spectral information into depth-resolved
information and the angular information into a transverse spatial
distribution. The capabilities of faLCI have been initially
demonstrated by extracting the size of polystyrene beads in a
depth-resolved measurement.
[0037] The key advances of the disclosed embodiments can be broken
down into three components: (1) new rapid data acquisition methods,
(2) fiber probe designs, and (3) data analysis schemes. Thus, the
disclosed embodiments are described in this matter for convenience
in its understanding.
[0038] An exemplary apparatus, as well as the steps involved in the
process of obtaining angle and depth-resolved distribution data
scattered from a sample, are also set forth in FIG. 2. The faLCI
scheme in accordance with one embodiment of the disclosed
embodiments is based on a modified Mach-Zehnder, interferometer as
illustrated in FIG. 1A. Broadband light 10 from a superluminescent
diode (SLD) 12 is directed by a mirror 13 (step 60 in FIG. 2) and
split into a reference beam 14 and an input beam 16 to a sample 18
by beamsplitter BS1 20 (step 62 in FIG. 3). The output power of the
SLD 12 may be 3 milliWatts, having a specification of .lamda.o=850
nm, .DELTA..lamda.=20 nm FWHM for example, providing sufficiently
low coherence length to isolate scattering from a cell layer within
tissue. The path length of the reference beam 14 is set by
adjusting retroreflector RR 22, but remains fixed during
measurement. The reference beam 14 is expanded using lenses L1 (24)
and L2 (26) to create illumination (step 64 in FIG. 2), which is
uniform and collimated upon reaching a spectrograph slit 48 in an
imaging spectrograph 29. For example, L1 may have a focal length of
1.5 centimeters, and L2 26 may have focal length of 15
centimeters.
[0039] Lenses L3 (31) and L4 (38) are arranged to produce a
collimated pencil beam 30 incident on the sample 18 (step 66 in
FIG. 2). By displacing lens L4 (38) vertically relative to lens L3
(31), the input beam 30 is made to strike the sample at an angle of
0.10 radians relative to the optical axis. This arrangement allows
the full angular aperture of lens L4 (38) to be used to collect
scattered light 40 from the sample f8. Lens L4 (38) may have a
focal length of 3.5 centimeters.
[0040] The light 40 scattered by the sample 18 is collected by lens
L4 (32) and relayed by a 4f imaging system comprised of lenses L5
(43) and L6 (44) such that the Fourier plane of lens L4 (32) is
reproduced in phase and amplitude at the spectrograph slit 48 (step
68 in FIG. 2). The scattered light 40 is mixed with the reference
field 14 at a second beamsplitter BS2 42 with the combined fields
46 falling upon the entrance slit (illustrated in FIG. 1B as
element 48) to the imaging spectrograph 29 (step 70 in FIG. 2). The
imaging spectrograph 29 may be the model SP2150i, manufactured by
Acton Research for example. FIG. 1B illustrates the distribution of
scattering angle across the dimension of the slit 48. The mixed
fields are dispersed with a high resolution grating (e.g. 1200
1/mm) and detected using a cooled CCD 50 (e.g. 1340.times.400, 20
.mu.m.times.20 .mu.m pixels, Spec10:400, manufactured by Princeton
Instruments) (step 72 in FIG. 2).
[0041] The detected signal 46 is a function of vertical position on
the spectrograph slit 48, .lamda., and wavelength once the light is
dispersed by the spectrograph 29. The detected signal at pixel (m,
n) can be related to the signal 40 and reference fields 16
(E.sub.s, E.sub.r,) as:
I(.lamda..sub.m,y.sub.n)=|E.sub.r(.lamda..sub.m,y.sub.n)|.sup.2+|E.sub.s-
(.lamda..sub.m,y.sub.n)|.sup.2+2ReE.sub.s(.lamda..sub.m,y.sub.n)E.sub.r*(.-
lamda..sub.m,y.sub.n)cos .phi., (1)
[0042] where .phi. is the phase difference between the two fields
30, 16 and . . . denotes an ensemble average in time. The
interference term is extracted by measuring the intensity of the
signal 30 and reference beams 16 independently and subtracting them
from the total intensity.
[0043] In order to obtain depth resolved information, the
wavelength spectrum at each scattering angle is interpolated into a
wavenumber (k=2.pi./.lamda.) spectrum and Fourier transformed to
give a spatial cross correlation, .GAMMA..sub.SR(z) for each
vertical pixel y.sub.n:
.GAMMA..sub.SR(z,y.sup.n)=.intg.dke.sup.zE.sub.s(k,y.sub.n)E.sub.r*(k,y.-
sub.n)cos .phi.. (2)
The reference field 14 takes the form
E.sub.r(k)=E.sub.oexp.left
brkt-bot.-((k-k.sub.o)/.DELTA.k).sup.2.right brkt-bot.exp.left
brkt-bot.-((y-y.sub.o)/.DELTA.y).sup.2.right
brkt-bot.exp[ik.DELTA.l] (3)
where k.sub.o (y.sub.o and .DELTA.k(.DELTA.y) represent the center
and width of the Gaussian wavevector (spatial) distribution and
.DELTA.l is the selected path length difference. The scattered
field 40 takes the form
E.sub.s(k,.theta.)=.SIGMA..sub.jE.sub.oexp[-((k-k.sub.o)/.DELTA.k).sup.2-
]exp[ikl.sub.j]S.sub.j(k,.theta.) (4)
[0044] where S.sub.j represents the amplitude distribution of the
scattering originating from the jth interface, located at depth
l.sub.j. The angular distribution of the scattered field 40 is
converted into a position distribution in the Fourier image plane
of lens L4 through the relationship y=f.sub.4.theta.. For the pixel
size of the CCD 50 (e.g. 20 .mu.m), this yields an angular
resolution (e.g. 0.57 mrad) and an expected angular range (e.g. 228
mrad.).
[0045] Inserting Eqs. (3) and (4) into Eq. (2) and noting the
uniformity of the reference field 14 (.DELTA.y>>>slit
height) yields the spatial cross correlation at the nth vertical
position on the detector 29:
.GAMMA. SR ( z , y n ) = j .intg. k E o 2 exp [ - 2 ( ( k - k 0 ) /
.DELTA. k ) 2 ] exp [ k ( z - .DELTA. l + l j ) ] .times. S j ( k ,
.theta. n = y n / f 4 ) cos .phi. . ( 5 ) ##EQU00001##
Evaluating this equation for a single interface yields:
.GAMMA..sub.SR(z,y.sub.n)=|E.sub.o|.sup.2exp[-((z-.DELTA.l+l.sub.j).DELT-
A.k).sup.2/8]S.sub.j(k.sub.o,.theta..sub.n=y.sub.n/f.sub.4)cos
.phi.. (6)
[0046] Here we have assumed that the scattering amplitude S does
not vary appreciably over the bandwidth of the source light 12.
This expression shows that we obtain a depth resolved profile of
the scattering distribution 40 with each vertical pixel
corresponding to a scattering angle.
[0047] FIG. 3A below shows typical data representing the total
detected intensity (Equation (1), above) of the sum of the
reference field 16 and the field scattered 40 by a sample of
polystyrene beads, in the frequency domain given as a function of
wavelength and angle, given with respect to the backwards
scattering direction. In an exemplary embodiment, this data was
acquired in 40 milliseconds and records data over 186 mrad,
approximately 85% of the expected range, with some loss of signal
at higher angles.
[0048] FIGS. 3B and 3C illustrate the intensity of the reference
and signal fields 14, 30 respectively. Upon subtraction of the
signal and reference fields 14, 30 from the total detected
intensity, the interference 46 between the two fields is realized
as illustrated in FIG. 3D. At each angle, interference data 46 are
interpolated into k-space and Fourier transformed to give the
angular depth resolved profiles of the sample 18 as illustrated in
FIG. 4A. The Fourier transform of the angle-resolved, cross
correlated signal 46, which is the result of signal 40 scattered at
a multitude of reflected angles off the sample 18 and obtained in
the Fourier plane of lens L4 (38), produces depth-resolved
information about the sample 18 as a function of angle and depth.
This provides depth-resolved information about the sample 18.
Because the angle-resolved, cross-correlated signal 46 is
spectrally dispersed, the data acquisition permits data to be
obtained in less than one second. Information about all depths of
the sample 18 at each of the multitude of different points (i.e.
angles) on the sample 18 can be obtained with one scan on the order
of approximately 40 milliseconds. Normally, time domain based
scanning is required to obtain information about all depths of a
sample at a multitude of different points, thus requiring
substantial time and movement of the reference arm with respect to
the sample.
[0049] In the experiments that produced the depth-resolved profile
of the sample 18 illustrated in FIG. 4A, the sample 18 consists of
polystyrene microspheres (e.g. n=1.59, 10.1 .mu.m mean diameter,
8.9% variance, NIST certified, Duke Scientific) suspended in a
mixture of 80% water and 20% glycerol (n=1.36) to provide neutral
buoyancy. The solution was prepared to obtain a scattering length
l=200 .mu.m. The sample is contained in a round well (8 mm
diameter, 1 mm deep) behind a glass coverslip (thickness,
d.about.170 .mu.m) (not shown). The sample beam 30 is incident on
the sample 18 through the coverslip. The round trip thickness
through the coverslip (2 n d=2(1.5)(170 .mu.m)=0.53 mm--see FIG.
4A) shows the depth resolved capability of the approach. The data
are ensemble averaged by integrating over one mean free path (MFP).
The spatial average can enable a reduction of speckle when using
low-coherence light to probe a scattering sample. To simplify the
fitting procedure, the scattering distribution is low pass filtered
to produce a smoother curve, with the cutoff frequency chosen to
suppress spatial correlations on length scales above 16 .mu.m.
[0050] In addition to obtaining depth-resolved information about
the sample 18, the scattering distribution data (i.e. a/LCI data)
obtained from the sample 18 using the disclosed data acquisition
scheme can also be used to make a size determination of the nucleus
using the Mie theory. A scattering distribution 74 of the sample 18
is illustrated in FIG. 4B as a contour plot. The raw scattered
information 74 about the sample 18 is shown as a function of the
signal field 30 and angle. A filtered curve is determined using the
scattered data 74. Comparison of the filtered scattering
distribution curve 76 (i.e. a representation of the scattered data
74) to the prediction of Mie theory (curve 78 in FIG. 5A) enables a
size determination to be made.
[0051] In order to fit the scattered data 76 to Mie theory, the
a/LCI signals are processed to extract the oscillatory component
which is characteristic of the nucleus size. The smoothed data 76
are fit to a low-order polynomial (4.sup.th order was used for
example herein, but later studies use a lower 2.sup.nd order),
which is then subtracted from the distribution 76 to remove the
background trend. The resulting oscillatory component is then
compared to a database of theoretical predictions obtained using
Mie theory 78 from which the slowly varying features were similarly
removed for analysis.
[0052] A direct comparison between the filtered a/LCI data 76 and
Mie theory data 78 may not be possible, as the chi-squared fitting
algorithm tends to match the background slope rather than the
characteristic oscillations. The calculated theoretical predictions
include a Gaussian distribution of sizes characterized by a mean
diameter (d) and standard deviation (ED) as well as a distribution
of wavelengths, to accurately model the broad bandwidth source.
[0053] The best fit (FIG. 5A) is determined by minimizing the
Chi-squared between the data 76 and Mie theory (FIG. 5B), yielding
a size of 10.2+/-1.7 .mu.m, in excellent agreement with the true
size. The measurement error is larger than the variance of the bead
size, most likely due to the limited range of angles recorded in
the measurement.
[0054] As an alternative to processing the a/LCI data and comparing
to Mie theory, there are several other approaches which could yield
diagnostic information. These include analyzing the angular data
using a Fourier transform to identify periodic oscillations
characteristic of cell nuclei. The periodic oscillations can be
correlated with nuclear size and thus will possess diagnostic
value. Another approach to analyzing a/LCI data is to compare the
data to a database of angular scattering distributions generated
with finite element method (FEM) or T-Matrix calculations. Such
calculations may offer superior analysis as there are not subject
to the same limitations as Mie theory. For example, FEM or T-Matrix
calculations can model non-spherical scatterers and scatterers with
inclusions while Mie theory can only model homogenous spheres.
[0055] As an alternative embodiment, the disclosed embodiments can
also employ optical fibers to deliver and collect light from the
sample of interest to use in the a/LCI system for endoscopic
applications. This alternative embodiment is illustrated in FIG.
6.
[0056] The fiber optic a/LCI scheme for this alternative embodiment
makes use of the Fourier transform properties of a lens. This
property states that when an object is placed in the front focal
plane of a lens, the image at the conjugate image plane is the
Fourier transform of that object. The Fourier transform of a
spatial distribution (object or image) is given by the distribution
of spatial frequencies, which is the representation of the image's
information content in terms of cycles per mm. In an optical image
of elastically scattered light, the wavelength retains its fixed,
original value and the spatial frequency representation is simply a
scaled version of the angular distribution of scattered light.
[0057] In the fiber optic a/LCI scheme, the angular distribution is
captured by locating the distal end of the fiber bundle in a
conjugate Fourier transform plane of the sample using a collecting
lens. This angular distribution is then conveyed to the distal end
of the fiber bundle where it is imaged using a 4f system onto the
entrance slit of an imaging spectrograph. A beamsplitter is used to
overlap the scattered field with a reference field prior to
entering the slit so that low coherence interferometry can also be
used to obtain depth resolved measurements.
[0058] Turning now to FIG. 6, the fiber optic faLCI scheme is
shown. Light 12' from a broadband light source 10' is split into a
reference field 14' and a signal field 16' using a fiber splitter
(FS) 80. A splitter ratio of 20:1 is chosen in one embodiment to
direct more power to a sample 18' via the signal arm 82 as the
light returned by the tissue is typically only a small fraction of
the incident power.
[0059] Light in the reference fiber 14' emerges from fiber F1 and
is collimated by lens L1 (84) which is mounted on a translation
stage 86 to allow gross alignment of the reference arm path length.
This path length is not scanned during operation but may be varied
during alignment. A collimated beam 88 is arranged to be equal in
dimension to the end 91 of fiber bundle F3 (90) so that the
collimated beam 88 illuminates all fibers in F3 with equal
intensity. The reference field 14' emerging from the distal tip of
F3 (90) is collimated with lens L3 (92) in order to overlap with
the scattered field conveyed by fiber F4 (94). In an alternative
embodiment, light emerging from fiber F1 (14') is collimated then
expanded using a lens system to produce a broad beam.
[0060] The scattered field is detected using a coherent fiber
bundle. The scattered field is generated using light in the signal
arm 82 which is directed toward the sample 18' of interest using
lens L2 (98). As with the free space system, lens L2 (98) is
displaced laterally from the center of single-mode fiber F2 such
that a collimated beam is produced which is traveling at an angle
relative to the optical axis The fact that the incident beam
strikes the sample at an oblique angle is essential in separating
the elastic scattering information from specular reflections. The
light scattered by the sample 18' is collected by a fiber bundle
consisting of an array of coherent single mode or multi-mode
fibers. The distal tip of the fiber is maintained one focal length
away from lens L2 (98) to image the angular distribution of
scattered light. In the embodiment shown in FIG. 6, the sample 18'
is located in the front focal plane of lens L2 (98) using a
mechanical mount 100. In the endoscope compatible probe shown in
FIG. 7, the sample is located in the front focal plane of lens L2
(98) using a transparent sheath (element 102).
[0061] As illustrated in FIG. 6 and also FIG. 7B, scattered light
104 emerging from a proximal end 105 of the fiber probe F4 (94) is
recollimated by lens L4 (104) and overlapped with the reference
field 14' using beamsplitter BS (108). The two combined fields 110
are re-imaged onto the slit (element 48' in FIG. 7) of the imaging
spectrograph 29' using lens L5 (112). The focal length of lens L5
(112) may be varied to optimally fill the slit 48'. The resulting
optical signal contains information on each scattering angle across
the vertical dimension of the slit 48' as described above for the
apparatus of FIGS. 1A and 1B.
[0062] It is expected that the above-described a/LCI fiber-optic
probe will collect the angular distribution over a 0.45 radian
range (approx. 30 degrees) and will acquire the complete depth
resolved scattering distribution 110 in a fraction of a second.
[0063] There are several possible schemes for creating the fiber
probe which are the same from an optical engineering point of view.
One possible implementation would be a linear array of single mode
fibers in both the signal and reference arms. Alternatively, the
reference arm 96 could be composed of an individual single mode
fiber with the signal arm 82 consisting of either a coherent fiber
bundle or linear fiber array.
[0064] The fiber probe tip can also have several implementations
which are substantially equivalent. These would include the use of
a drum or ball lens in place of lens L2 (98). A side-viewing probe
could be created using a combination of a lens and a minor or prism
or through the use of a convex minor to replace the lens-minor
combination. Finally, the entire probe can be made to rotate
radially in order to provide a circumferential scan of the probed
area.
[0065] Yet another data acquisition embodiment of the disclosed
embodiments could be a fa/LCI system is based on a modified
Mach-Zehnder interferometer as illustrated in FIG. 5A. The output
10'' from a fiber-coupled superluminescent diode (SLD) source 12''
(e.g. Superlum, P.sub.o=15 mW, .lamda.o=841.5 nm,
.DELTA..lamda.=49.5 nm, coherence length=6.3 .mu.m) is split into
sample arm delivery fiber 16'' and a reference arm delivery fiber
14'' by a 90/10 fiber splitter FS (80') (e.g. manufactured by AC
Photonics). The sample arm delivery fiber 16'' can consist of
either of the following for example: (1) a single mode fiber with
polarization control integrated at the tip; or (2) a polarization
maintaining fiber. A sample probe 113 is assembled by affixing the
delivery fiber 16''(NA.apprxeq.0.12) along the ferrule 114 at the
distal end of a fiber bundle 116 such that the end face of the
delivery fiber 16'' is parallel to and flush with the face of the
fiber bundle 116. Ball lens L1 (115) (e.g. f=2.2 mm) is positioned
one focal length from the face of the probe 113 and centered on the
fiber bundle 116, offsetting the delivery fiber 16'' from the
optical axis of lens L1 (115). This configuration, which is also
depicted in FIG. 8B, produces a collimated beam 120 (e.g. P=9 mW)
with a diameter (e.g. 2f.sub.1NA) of 0.5 mm incident on the sample
18'' at an angle of 0.25 rad. for example.
[0066] The scattered light 122 from the sample is collected by lens
L1 (115) and, via the Fourier transform property of the lens L1
(115), the angular distribution of the scattered field 122 is
converted into a spatial distribution at the distal face of the
multimode coherent fiber bundle 116 (e.g. Schott North America,
Inc., length=840 mm, pixel size=8.2 .mu.m, pixel count=13.5K) which
is located at the Fourier image plane of lens L1 (115). The
relationship between vertical position on the fiber bundle, y', and
scattering angle, .theta. is given by y'=f.sub.1.theta.. As an
illustration, the optical path of light scattered 122 at three
selected scattering angles is shown in FIG. 8B. Overall, the
angular distribution is sampled by approximately 130 individual
fibers for example, across a vertical strip of the fiber bundle
116'', as depicted by the highlighted area in FIG. 8C. The 0.2 mm,
for example, thick ferrule (d.sub.1) separating the delivery fiber
16'' and fiber bundle 116 limits the minimum theoretical collection
angle (.theta..sub.min,th=d.sub.1/f.sub.1) to 0.09 rad in this
example. The maximum theoretical collection angle is determined by
d.sub.1 and d.sub.2, the diameter of the fiber bundle, by
.theta..sub.max,th=(d.sub.1+d.sub.2)/f.sub.1 to be 0.50 rad.
Experiments using a standard scattering sample 122 indicate the
usable angular range to be .theta..sub.min=0.12 rad. to
.theta..sub.max=0.45 rad. d.sub.1, for example, can be minimized by
fabricating a channel in the distal ferrule 123 and positioning the
delivery fiber 16'' in the channel. The fiber bundle 116 is
spatially coherent, resulting in a reproduction of the collected
angular scattering distribution at the proximal face. Additionally,
as all fibers in the bundle 116 are path length matched to within
the coherence length, the optical path length traveled by scattered
light 122 at each angle is identical. The system disclosed in
"Fiber-optic-bundle-based optical coherence tomography," by T. Q.
Xie, D. Mukai, S. G. Guo, M. Brenner, and Z. P. Chen in Optics
Letters 30(14), 1803-1805 (2005) (hereinafter "Xie"), incorporated
by reference herein in its entirety, discloses a multimode coherent
fiber bundle into a time-domain optical coherence tomography system
and demonstrated that the modes of light coupled into an individual
fiber will travel different path lengths. In the example herein of
the disclosed embodiments, it was experimentally determined that
the higher order modes are offset from the fundamental mode by 3.75
mm, well beyond the depth (.about.100 .mu.m) required for gathering
clinically relevant data. Additionally, the power in the higher
order modes had a minimal effect on dynamic range as the sample arm
power is significantly less than the reference arm power. Finally,
it should be noted that while the system disclosed in Xie collected
data serially through individual fibers, the example of the
disclosed embodiments herein uses 130 fibers to simultaneously
collect scattered light across a range of angles in parallel,
resulting in rapid data collection.
[0067] The angular distribution exiting a proximal end 124 of the
fiber bundle 116 is relayed by the 4f imaging system of L2 and L3
(f.sub.2=3.0 cm, f.sub.3=20.0 cm) to the input slit 48'' of the
imaging spectrograph 29'' (e.g. Acton Research, InSpectrum 150).
The theoretical magnification of the 4f imaging system is
(f.sub.3/f.sub.2) 6.67 in this example. Experimentally, the
magnification was measured to be M=7.0 in this example with the
discrepancy most likely due to the position of the proximal face
124 of the fiber bundle 116 with relation to lens L2 (126). The
resulting relationship between vertical position on the
spectrograph slit 48'', y, and .theta. is
y=Mf.sub.1(.theta.-.theta..sub.min). The optical path length of the
reference arm is matched to that of the fundamental mode of the
sample arm. Light 127 exiting the reference fiber 14'' is
collimated by lens L4 (128) (e.g. f=3.5 cm, spot size=8.4 mm) to
match the phase front curvature of the sample light and to produce
even illumination across the slit 48'' of the imaging spectrograph
29''. A reference field 130 may be attenuated by a neutral density
filter 132 and mixed with the angular scattering distribution at
beamsplitter BS (134). The mixed fields 136 are dispersed with a
high resolution grating (e.g. 1200 lines/mm) and detected using an
integrated, cooled CCD (not shown) (e.g. 1024.times.252, 24
.mu.m.times.24 .mu.m pixels, 0.1 nm resolution) covering a spectral
range of 99 nm centered at 840 nm, for example.
[0068] The detected signal 136, a function of wavelength, 2, and 0,
can be related to the signal and reference fields (Es, Er) as:
I(.lamda..sub.m,.theta..sub.n)=|E.sub.r(.lamda..sub.m,.theta..sub.n|.sup-
.2+|E.sub.s(.lamda..sub.m,.theta..sub.n)|.sup.2+2ReE.sub.s(.lamda..sub.m,.-
theta..sub.n)E.sub.r*(.lamda..sub.m,.theta..sub.n)cos(.phi.),
(1)
where .phi. is the phase difference between the two fields, (m,n)
denotes a pixel on the CCD, and . . . denotes a temporal average.
I(.lamda..sub.m,.theta..sub.n) is uploaded to a PC using LabVIEW
manufactured by National Instruments software and processed in 320
ms to produce a depth and angle resolved contour plot of scattered
intensity. The processing of the angle-resolved scattered field to
obtain depth and size information described above, and in
particular reference to the data acquisition apparatus of FIGS. 1A
and 1B, can then be used to obtain angle-resolved, depth-resolved
information about the sample 18'' using the scattered mixed field
136 generated by the apparatus in FIG. 8.
[0069] The embodiments set forth above represent the necessary
information to enable those skilled in the art to practice the
disclosed embodiments and illustrate the best mode of practicing
the disclosed embodiments. Upon reading the following description
in light if the accompanying drawings figures, those skilled in the
art will understand the concepts of the disclosed embodiments and
will recognize applications of these concepts not particularly
addressed herein. It should be understood that these concepts and
applications fall within the scope of the disclosure.
[0070] Those skilled in the art will recognize improvements and
modifications to the preferred embodiments of the disclosed
embodiments. All such improvements and modifications are considered
within the scope of the concepts disclosed herein and the claims
that follow.
* * * * *