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Title:
DIFFRACTION-GRATING-BASED COMMON-PATH INTERFEROMETER FOR IMAGING FOURIER-TRANSFORM SPECTROSCOPY
Document Type and Number:
WIPO Patent Application WO/2016/115321
Kind Code:
A2
Abstract:
An interferometer employing a 4f optical imaging system, through which both reference and sample beams are propagated, between diffraction gratings configured as input and output beam-splitting / beam-combining components. A Fourier-transform spectrometer utilizing the same and a microscope as an input optical sub-system. The interferometer includes a variable-phase-delay optical element in a Fourier plane of the imaging spectrometer defined between lens elements of the 4f optical system. In a special case, an additional 4f optical imaging system is used at the output of the interferometer, through which interferograms are registered at the detector. Light output collected by the detector has the same optical path difference between reference and sample beams at any point across field-of-view. A method for performing imaging spectrometry and forming images of an object under the microscope.

Inventors:
WADDUWAGE DUSHAN NAWODA (SG)
SINGH VIJAY RAJ (SG)
CHOI HEEJIN (US)
YAQOOB ZAHID (US)
SO PETER (US)
Application Number:
PCT/US2016/013366
Publication Date:
July 21, 2016
Filing Date:
January 14, 2016
Export Citation:
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Assignee:
MASSACHUSETTS INST TECHNOLOGY (US)
UNIV SINGAPORE (SG)
International Classes:
G01J3/453; G01B9/02
Other References:
KORENBERG, M J ET AL.: "Raman spectral estimation via fast orthogonal search", ANALYST, vol. 122, no. 9, 1997, pages 879 - 882, XP002092858, DOI: doi:10.1039/a700902j
KORENBERG ET AL.: "Raman spectral estimation via fast orthogonal search", ANALYST, vol. 122, no. 9, 1997, pages 879 - 882, XP002092858, DOI: doi:10.1039/a700902j
Attorney, Agent or Firm:
SIDORIN, Yakov (One South Church Avenue Suite 170, Tucson AZ, US)
Download PDF:
Claims:
CLAIMS

What is claimed is:

1. A common-path optical interferometer comprising:

a first lens system including first and second lens elements separated, along an axis of the interferometer, by a distance substantially equal to a sum of the first and second focal lengths, said first and second focal lengths being respective focal lengths of said first and second lenses, said first lens system defining an only arm of said interferometer;

an input beam-splitter disposed in front of said first lens system and separated therefrom by the first focal length; and

an output beam-splitter disposed behind said first lens system at a distance equal to the second focal length;

wherein each of said beam-splitters is configured to form first and second diffracted beams from an optical wave incident thereon, the first and second diffracted beams inclined at different angles with respect to the axis.

2. An interferometer according to claim 1, wherein the first and second diffracted beams have the same order of diffraction.

3. An interferometer according to claim 1, wherein reference and signal beams of light propagating through said only arm are defined, respectively, by said first and second diffracted beams.

4. An interferometer according to claim 1, further comprising at least one phase-delay component positioned across at least one of said first and second diffracted beams at a first Fourier transform plane defined by the first lens element, said at least one phase-delay component structured to vary a phase-delay introduced into the at least one of said first and second diffracted beams, as a result of spatial reorientation of said component.

5. An interferometer according to claim 4, wherein the variable phase-delay optical element includes first and second optical prismatic elements having respectively -corresponding portions with first and second shapes that are complementary to one another.

6. An interferometer according to claim 4, wherein the variable phase-delay optical element includes a slab of optical material, the variable phase-delay optical element configured to rotate said slab about an axis that is perpendicular to the axis of the 4f optical system.

7. An interferometer according to claim 4, wherein the variable phase-delay optical element includes a step-phase plate of optical material, the variable phase-delay optical element configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system.

8. An interferometer according to claim 1, further comprising a second lens system including third and forth lens elements disposed to receive light from the output beam-splitter, to form a plane between the third and fourth elements in which said light is focused.

9. An interferometer according to claim 1, further comprising an input optical system defining a first intermediate image plane, in light passing therethrough, at the input beam-splitter.

10. An imaging spectrometer comprising an interferometer according to claim 1, and further comprising

at least one phase-delay component positioned across at least one of said first and second diffracted beams at a first Fourier transform plane defined by the first lens element;

a second lens system including third and fourth lens elements separated, along the axis, by a distance substantially equal to a sum of the third and fourth focal lengths, said third and fourth focal lengths being respective focal lengths of said third and fourth lenses; and

an optical detector disposed to receive light that has propagated through the first and second lens systems.

11. An imaging spectrometer according to claim 10, having a uniform distribution of phase of light, received by the optical detector, across a field of view (FOV) of the optical detector.

12. An imaging spectrometer comprising:

an input optical system defining a first intermediate image plane in light passing therethrough; and

a common-path interferometer including a 4f optical system positioned to form an optical conjugate of the first intermediate image plane in a second intermediate image plane that is located at a focal distance behind the 4f optical system, the focal distance being equal to a focal length of a lens element of the 4f optical system;

a first transmission diffraction grating positioned in the first intermediate image plane perpendicularly to an axis of the 4f optical system; and

a variable phase-delay optical element in a Fourier plane defined by the 4f optical system between lens elements of the 4f optical system.

13. An imaging spectrometer according to claim 12, further comprising

a second transmission grating positioned in the second intermediate image plane perpendicularly to the axis of the 4f optical system; and

an auxiliary 4f optical system positioned co-axially with the 4f optical system to form an optical conjugate of the second intermediate image plane at a location separated from the auxiliary 4f optical system by a focal length of a lens element of the 4f auxiliary optical system; and

an optical detector positioned in such location that light received by the optical detector from the first diffraction grating has a uniform distribution.

14. An imaging spectrometer according to claim 12, wherein the variable phase-delay optical element includes first and second optical prismatic elements having respectively-corresponding portions with first and second shapes that are complementary to one another.

15. An imaging spectrometer according to claim 12, wherein the variable phase-delay optical element includes a slab of optical material, the variable phase-delay optical element configured to rotate said slab about an axis that is perpendicular to the axis of the 4f optical system.

16. An imaging spectrometer according to claim 12, wherein the variable phase-delay optical element includes a step-phase plate of optical material, the variable phase-delay optical element configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system.

17. An imaging spectrometer according to claim 12, further comprising an optical detector disposed in the second intermediate imaging plane to receive light that has propagated along the axis of the 4f optical system; wherein light received at the optical detector has a sinusoidally-modulated distribution of phase across a field of view (FOV) of the optical detector.

18. A common-path optical interferometer comprising:

a transmission diffraction grating disposed in a first intermediate image plane;

a 4f optical system having an axis and positioned to form an optical conjugate of the first intermediate image plane in a second intermediate image plane that is located at a focal distance behind the 4f optical system, the focal distance being substantially equal to a focal length of a lens element of the 4f optical system;

a variable phase-delay optical element in a Fourier plane defined by the 4f optical system between lens elements of the 4f optical system,

wherein a distribution of phase of a collimated beam of light, incident along the axis into the transmission diffraction grating and transmitted through the 4f optical system, is sinusoidally-modulated across a field of view (FOV) defined by said light upon transmission through the 4f imaging system at the second intermediate image plane.

19. An interferometer according to claim 18, wherein the variable phase-delay optical element includes first and second optical prismatic elements having respectively-corresponding portions with first and second shapes that are complementary to one another.

20. An interferometer according to claim 18, wherein the variable phase-delay optical element includes a slab of optical material, the variable phase-delay optical element configured to rotate said slab about an axis that is perpendicular to the axis of the 4f optical system.

21. An interferometer according to claim 18, wherein the variable phase-delay optical element includes a step-phase plate of optical material, the variable phase-delay optical element configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system.

22. An imaging spectrometer comprising an interferometer according to claim 18, further comprising an optical detector positioned across the axis in a plane that is optically conjugate to the first intermediate image plane as defined in transmission of light through the 4f optical system.

23. A method for performing imaging spectrometry, the method comprising: propagating a beam of light, delivered with an optical system from an object, through a common- path interferometer including a 4f optical system; and

acquiring light transmitted through the common-path interferometer with an optical detector to determine spectral distribution of said light.

24. A method according to claim 23, further comprising defining a distribution of phase of light from said beam of light that has propagated through said common-path interferometer, to be sinusoidally- modulated across a field of view (FOV) of the optical detector.

25. A method according to claim 23, wherein said propagating includes forming first and second diffracted beams from the beam of light transmitted through a first diffraction grating disposed in a first plane;

propagating the first and second diffracted beams through the 4f optical system to transmit light from the first and second diffracted beams through a second diffraction grating disposed in a second plane, wherein the second plane is optically-conjugate to the first plane in light propagated through the 4f optical system.

26. A method according to claim 25, further comprising

transmitting light from a collimated beam of light, that has propagated through the 4f optical system, through an auxiliary 4f optical system positioned co-axially with the 4f optical system to form an interferogram at the optical detector, said optical detector being disposed at a plane that is optically- conjugate to the first plane.

27. A method according to claim 25, further comprising defining a distribution of phase of light acquired by the optical detector, to be uniform across a field of view (FOV) of the optical detector.

28. A method according to claim 23, further comprising optically delaying one of the first and second diffracted beams with a spatially-repositionable variable phase-delay optical element disposed in a Fourier transform plane, the Fourier transform plane defined by a lens element of the 4f optical system with respect to said collimated beam of light.

29. A method according to claim 28, wherein said optically delaying includes disposing first and second optical prismatic element across one of the first and second diffracted beams, said firs and second optical prisms having respectively-corresponding portions with first and second shapes that are complementary to one another.

30. A method according to claim 28, wherein said optically delaying includes disposing a slab of optical material across one of the first and second diffracted beams, and wherein the variable phase-delay optical element is configured to rotate said slab about an axis that is perpendicular to the axis of the 4f optical system.

31. A method according to claim 28, wherein said optically delaying includes positioning a step- phase plate of optical material across one of the first and second diffracted beams, and wherein the variable phase-delay optical element is configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system

32. A method for measuring a spectral characteristic at all pixels of an image of a two-dimensional scene, the method comprising:

for each of a plurality of chosen light sources, forming a respectively-corresponding

interferogram at an output of an imaging spectrometer, wherein the imaging spectrometer is configured to produce, at the output, a distribution of light having uniform distribution of phase across a field-of-view (FOV) corresponding to said output, and wherein each of the plurality of chosen light source has a known spectral signature;

with an optical detector, randomly sampling each of formed interferograms with varying number of samples and optical-path-difference (OPD) values to obtain optical data, the OPD values representing phase delays of light propagating through the imaging spectrometer towards the output;

fitting said optical data to respectively -corresponding known spectral signatures of the plurality of chosen light source to calculate values representing accuracies of said fitting;

selecting OPD values corresponding to the smallest number of samples that satisfy a threshold accuracy of fitting; and

performing interference measurements of the two-dimensional scene for each of selected OPD values to determine the spectral characteristic.

33. A method according to claim 32, further comprising varying the OPD values, representing values of OPD between reference and signal beams of an interferometer of said imaging spectrometer, wherein said varying the OPD values is not random.

34. A method according to claim 32, further comprising transmitting light through an interferometer of said imaging spectrometer, wherein the interferometer is a common-path interferometer.

35. A method according to claim 34, further comprising non-randomly changing said ODP values, wherein the OPD values are values of OPD between reference and signal beams of said common-path interferometer.

Description:
DIFFRACTION-GRATING-BASED COMMON-PATH INTERFEROMETER FOR IMAGING FOURIER-TRANSFORM SPECTROSCOPY

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims priority from and benefit of the US Provisional Patent

Application No. 62/125,279 filed on January 16, 2015 and titled "Diffraction Grating Based Common Path Interferometer for Imaging Fourier Transform Spectroscopy", the disclosure of which is incorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was partially made with government support under Grant Number

P41EB015871 awarded by the National Institute of Health. The U.S. govemment has certain rights in the invention.

TECHNICAL FIELD

[0003] The present invention relates to imaging spectroscopy and, more particularly, to Fourier- transform spectroscopy apparatus and method employing a grating -based common-path interferometer.

BACKGROUND

[0004] Imaging Fourier transform spectroscopy (FTS) is a wide-field imaging spectroscopic technique used for hyperspectral imaging. In implementing the FTS, an interferometer is used to measure interference patterns for every pixel in the field of view (FOV) simultaneously, at controlled optical path differences (OPDs). The interferometer used for FTS measurements should satisfy at least two criteria. First, it should be stable enough (in terms of phase of the optical signal) to allow for measurement of optical interference produced by optically-weak signals. The Sagnac interferometer is considered to be the industry standard in this respect because of its common-path design providing for excellent phase stability during the measurements. The second requirement - which the Sagnac interferometer does not necessarily satisfy - is the requirement of phase uniformity (PU) throughout the FOV of the device, which is required for high- throughput applications. The importance of PU is evident when FTS is used with down-sampled algorithms such as fast orthogonal search, for example (which use may be required for data- acquisition at higher speeds. SUMMARY

[0005] An embodiment of the invention provides a common-path optical interferometer and a

Fourier-transform spectrometer utilizing the same. The interferometer includes a first lens system having first and second lens elements separated, along an axis of the interferometer, by a distance equal to a sum of the first and second focal lengths. The spectrometer includes an input optical system (such as a microscope) at the input of the interferometer to provide imaging of an object in a collimated beam delivered by the microscope to the interferometer. Here, the first and second focal lengths are respective focal lengths of the first and second lenses. This first lens system defining a single, only arm of the interferometer. Interferometer further includes an input beam-splitter disposed in front of said first lens system and separated from it by the first focal length; and an output beam-splitter / beam-combiner disposed behind the first lens system at a distance equal to the second focal length. Each of these beam-splitting / beam-combining elements is configured to form first and second diffracted beams from an optical plane wave incident on such element, such that the first and second diffracted beams have the same order of diffraction. In one case, the reference and signal beams of light propagating through such single, only arm are defined, respectively, by the first and second diffracted beams. The input optical system is positioned such as to define a first intermediate image plane, in light passing therethrough, at the input beam-splitter.

[0006] An interferometer may further include at least one phase-delay component positioned across at least one of the first and second diffracted beams at a first Fourier transform plane, which plane is defined by the first lens element in the incident optical plane wave. Such at least one phase-delay component is structured to vary a phase-delay introduced into the at least one of the first and second diffracted beams, as a result of spatial reorientation of this phase-delay component. Depending on the specifics of the

implementation, the variable phase-delay optical element includes either (a) first and second optical prisms having respectively-corresponding portions with first and second shapes that are complementary to one another; or (b) a slab of optical material, the variable phase-delay optical element configured to rotate said slab about an axis that is perpendicular to the axis of the 4f optical system; or (c) a step-phase plate of optical material, the variable phase-delay optical element configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system. Alternatively or in addition, the interferometer may further include a second lens system with third and fourth lens elements separated, along the axis, by a distance equal to a sum of the third and fourth focal lengths, said third and fourth focal lengths being respective focal lengths of said third and fourth lenses. An optical detector, disposed to receive light that has propagated through the first and second lens systems, received such light with a uniform distribution of phase of light across a field of view (FOV) of the optical detector.

[0007] Embodiments of the invention also provide an imaging spectrometer that includes an input optical system defining a first intermediate image plane in light passing therethrough; and a common-path single-pass interferometer. A common-path interferometer contains (i) a 4f optical system positioned to form an optical conjugate of the first intermediate image plane in a second intermediate image plane that is located at a focal distance behind the 4f optical system, the focal distance being equal to a focal length of a lens element of the 4f optical system; (ii) first and second transmission diffraction gratings positioned in the first and second intermediate image planes perpendicularly to an axis of the 4f optical system; and (iii) a variable phase-delay optical element in a Fourier plane defined by the 4f optical system between lens elements of the 4f optical system. In a specific case, the imaging spectrometer may also include an auxiliary 4f optical system positioned co-axially with the 4f optical system, at the output of it, to form an optical conjugate of the second intermediate image plane at a location separated from the auxiliary 4f optical system by a focal length of a lens element of the 4f auxiliary optical system. The variable-phase-delay optical element can be configured in ways referred to above. An optical detector, disposed to receive light that has propagated along the axis of the 4f optical system and the auxiliary 4f optical system, records interferograms in light that has the same optical path difference between the reference and sample beams of the interferometer for every point across the FOV.

[0008] Embodiments of the invention additionally provide a common-path optical interferometer that includes a transmission diffraction grating disposed in a first intermediate image plane; and a 4f optical system having an axis and positioned to form an optical conjugate of the first intermediate image plane in a second intermediate image plane that is located at a focal distance behind the 4f optical system, where the focal distance is equal to a focal length of a lens element of the 4f optical system. The interferometer also includes a variable phase-delay optical element in a Fourier plane defined by the 4f optical system between lens elements of the 4f optical system and is configured to form, from a collimated beam of light incident along the axis into the transmission diffraction grating and transmitted through the 4f optical system, a distribution of light the phase of which is sinusoidally-modulated across a field of view (FOV) defined by such light upon transmission through the 4f imaging system. The variable phase-delay optical element may be implemented in any of the forms mentioned above. In a specific implementation of such interferometer an imaging spectrometer is formed to include the interferometer and an optical detector positioned across the axis in a plane that is optically conjugate to the first intermediate plane as defined in transmission of light through the 4f optical system. [0009] Embodiments further provide a method for performing imaging spectrometry. The method includes the steps of (i) propagating light from a collimated beam of light, delivered with a microscope from an object, through a common-path interferometer including a 4f optical system; and (ii) acquiring light transmitted through the common-path interferometer with an optical detector to determine spectral distribution of such light. The method may further include a step of defining a distribution of phase of light, which has propagated through said common-path interferometer, to be sinusoidally-modulated across a field of view (FOV) of the optical detector. In particular, the method may include a step of optically delaying one of the first and second diffracted beams with a spatially-repositionable variable phase-delay optical element disposed in a Fourier transform plane, which plane is defined a lens element of the 4f optical system with respect to said collimated beam of light. In a specific case, the process of optically delaying includes positioning a step-phase plate of optical material across one of the first and second diffracted beams, and wherein the variable phase-delay optical element is configured to rotate the step-phase plate about an axis of said plate, where the axis of the plate is parallel to the axis of the 4f optical system.

[0010] The step of propagating includes, in one implementation, forming first and second diffracted beams from the collimated beam of light transmitted through a first diffraction grating disposed in a first plane; and propagating the first and second diffracted beams through the 4f optical system to transmit light from the first and second diffracted beams through a second diffraction grating disposed in a second plane. Here, the second plane is defined as being optically-conjugate to the first plane in light propagated through the 4f optical system.

[0011] In such implementation the method additionally includes transmitting light from a collimated beam of light (that has already propagated through the 4f optical system) through an auxiliary 4f optical system positioned co-axially with the 4f optical system to form an interferogram at the optical detector disposed at a plane that is optically-conjugate to the first plane. Alternatively or in addition, such implementation may involve defining a distribution of phase of light acquired by the optical detector, to be uniform across a field of view (FOV) of the optical detector.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The invention will be more fully understood by referring to the following Detailed

Description of Specific Embodiments in conjunction with the generally not-to scale Drawings, of which:

Figs. 1A, IB, and 1C schematically illustrate Fourier Transform Spectrometry methodology according to the present invention. Fig. 1A depicts a logical schematic of an embodiment of the system containing a common-path interferometer that is configured according to the idea of the invention; Fig. IB provides an illustration of the process of recovery of spectra according to the proposed methodology; Fig. 1C shows examples of images of an object recovered with the embodiments of Figs. 1A, IB in different spectral regions (indicated by four dominant wavelengths);

Fig. 2A is a schematic representation of an optical train of the imaging spectrometer of the invention that employs an embodiment of the common-path interferometer of the invention;

Fig. 2B is a schematic diagram of the input optical sub-system preceding the common-path interferometer of the embodiment of Fig. 2A;

Fig. 3 is a schematic diagram of prism(s) for use with embodiments of the common-path interferometer and the FTS utilizing such interferometer, to enable prism-based scanning of OPD;

Fig. 4 is a schematic diagram illustrating related methodology of implementing the OPD scanning in embodiments of the invention;

Figs. 5A, 5B, 5C illustrate plan top and plan side views of an embodiment of a phase-plate configured to achieve the incremental variation of the OPD between the reference and sample beams inside the common-path interferometer of the embodiment of Fig. 2A;

Fig. 6 illustrates the use of a combination (stacked) step-phase plates configured for an extended range OPD scanning;

Figs 7A, 7B, 7C, 7D illustrate the results of determination of output spectra from interferograms acquired by the optical detector of the system for different types of illumination and/or objects;

Figs. 8A, 8B, 8C, and 8D provide images and plots representing phase-stability of the interferometer of Fig. 2A.

Figs. 9A, 9B, 9C, 9D, and 9E illustrate data comparing phase-uniformity of operation of various interferometers, signifying the value of use of such interferometers in FTS measurements. Fig. 9A: FOV pattern in Michelson interferometer; Fig. 9B: FOV pattern in the proposed common-path interferometer of Fig. 2A; Fig. 9C: FOV pattern in Sagnac interferometer; Fig. 9D: Experimental image of the object in the FOV of proposed system of Fig. 2A; Fig. 9E: Experimental image of an object obtained with a Sagnac- interferometer-based spectrometer;

Fig. 10 is a schematic diagram of an optical train of the imaging spectrometer of the invention that employs a related embodiment of the common-path interferometer of the invention;

Figs. 11A, 11B, l lC, 11D illustrate the results of simulated measurement of peak wavelength for a quantum dot (peak wavelength range 450 nm - 850 nm) using an IFTS system and Fast Orthogonal Search methodology (FOS). Fig. 11A: Full interferogram for the emission spectrum of a representative quantum dot with a peak wavelength of 660 nm. N measurements were taken (first randomly -see Fig. 11C, then optimally -see Fig. 1 ID) and used in FOS to find the most dominant wavelength. Fig. 1 IB: The spectrum of the quantum in Fig. 11A. Here FOS recovers only the peak wavelength not the full spectrum and therefore the number of measurements and hence the measurement time can be extremely low. Fig. 11C: The measurement error vs. N (i.e. the number of measurements) when the measurements' OPD positions were chosen randomly. Shown in graph are the worst case, average case and best case. The broad difference vs. the best and average case suggests that there are optimized OPD position sets that lead to better results. Fig. 1 ID: The measurement error vs. N in the presence of Poisson noise for the most optimum OPD position set. Number of photos per measurement: curve a-100, curve b - 1000, curve c -10,000. T his suggests that despite Poisson noise there are IFTS configurations that can measure peak wavelength of a quantum dot (in 450 nm- 850 nm range) using less than 2000 photons to an accuracy of lnm;

Figs. 12A, 12B, 12C, 12D, 12E, 12F illustrate recovery of peak wavelengths' values with the use of FOS, for 6 micron fluorescent beads (ex/em = 503/511 nm). The recovery has been accomplished with the sued of complete Nyquist sampled interferogram (Figs. 12A, 12D), with the use of optimized compressive sampled interferogram with 20% samples (Figs. 12B, 12E), and with the use of optimized compressive sampled interferogram with 5% samples (Figs. 12C, 12F). Here, Figs. 12A, 12B, 12C are intensity weighted true color images and Figs. 12D, 12E, 12F are their respective non-weighted counterparts;

Figs. 13A, 13B, 13C, 13D, 13E, 13F illustrate recovery of peak wavelengths' values with the use of FOS, for mouse muscle tissue sample with regenerated cells;

Figs. 14A, 14B illustrate average measurement error (in nm) with random sampling (curves 1, 2, 3 in Fig. 14B) and optimized sampling (curve 4) for mouse muscle tissue sample with regenerated cells, for varying compression ratios. Here, «=number of compressive measurements and N=number of Nyquist measurements. Fig. 14A shows the pixels over which the peak wavelength measurements were averaged.

Generally, the sizes and relative scales of elements in Drawings may be set to be different from actual ones to appropriately facilitate simplicity, clarity, and understanding of the Drawings. For the same reason, not all elements present in one Drawing may necessarily be shown in another.

DETAILED DESCRIPTION

[0013] In accordance with examples of embodiments of the present invention, methods and apparatus are disclosed for performing Fourier Transform Spectroscopy measurements. The proposed methodology relies on implementation of a specific common-path interferometer in which the input and output beam-splitters are formed by diffraction gratings while the optical train between the beamsplitters includes a ^/ " imaging system to achieve both the uniform phase (corresponding to the uniformity of the OPD between the reference and signal beams of the interferometer) at any point across the FOV and,

simultaneously, to reduce the susceptibility of the FTS system to unexpected or random phase-shifts between the reference and signal beams. [0014] Fourier transform spectroscopy is a powerful method to resolve complex spectral signals, especially those received from unknown endogenous molecules. The FTS system can be operated in a wide- field mode, which facilitates its use with many high-throughput, high-content imaging systems. In FTS, an interferometer is introduced in the optical path for scanning of the value of the optical path difference (OPD) between the signal and reference beams, and the Fourier transform of the interferogram acquired in light transmitted through the OPD scanning element represents the spectrum of light for each point.

[0015] There are a number of limitations in existing wide field FTS designs. A design based on the

Michelson interferometer, for example, is straightforward and the OPD is uniform across the whole field of view. Such design allows efficient spectral recovery of the information based on strategically-defined sampling at very few OPD values. However, a Michelson-based design (being a double-path design) is highly sensitive to phase-shifts or length changes between the reference and sample arms and at least for that reason cannot be used for sensitive measurements of low-intensity and low-coherent signals such as fluorescence emissions, for example.

[0016] An alternative design employing a Sagnac interferometer is very stable and commonly used.

At the same time, however, the Sagnac design is known to have different OPD values corresponding to different locations in the image. In other words, the FTS system based on the Sagnac design produces an inherent phase tilt, resulting in linear variation of the OPD varies across the FOV. This, in rum, inevitably requires the user (or an applicable automated system) to perform a sequential scan over a large range of OPD values, which makes this implementation practically incompatible with high-throughput imaging.

[0017] Embodiments of the present invention provide a novel common-path interferometer design for use in full-field FTS measurement, which design is characterized by phase stability comparable with that of the Sagnac-interferometer-based FTS system and by uniform OPD, thereby allowing strategic sampling of OPD values for spectral recovery, comparable to that of the Michelson-based design.

[0018] Figs. 1A, IB, 1C provide a schematic illustration of the proposed methodology.

Specifically, with the use of the FTS system of the invention that utilizes an embodiment of the proposed common-path interferometer (Fig. 1A), the optical detection unit (that includes an optical detector and electronic circuitry / processor configured to acquire optical data from the detector and implement a judiciously chosen data-processing algorithm to extract the spectral information contained in recorded optical images) analyzes multiple images of the object taken at different OPD values between the reference and signal beams of the interferometer and derives the frequency spectrum of the object based on distribution of the Interferograms at respective OPD values across the FOV of the system; Fig. IB.

[0019] Implementations of the present invention provide solutions to the persisting problem caused in FTS measurement systems of related art by the presence of phase-instability and/or non-uniformity of the OPD (between the signal and reference beams) throughout the FOV. The solution is implemented by combining a 4/-optical -system based common-path interferometer (in which the reference and signal beams are formed by a first diffraction grating configured as an input optical beamsplitter while an OPD between the reference and signal beams is introduced upon propagation of one of the diffracted beams to the second diffraction grating configured as an output optical beamsplitter / combiner) with another 4f optical system forming an FTS image at the final image plane. A term 4f optical system (or a similar term) for the purposes of this application refers to and denotes an optical system that includes two lens elements positioned such as to establish two optical conjugates one of which is located at a first focal distance in front of the optical system (the first focal distance being equal to the focal length of the first lens element) and another of which is located at a second focal distance behind the optical system (the second focal distance being equal to the focal length of the second of the two lens elements). As understood in the field of optics, an object point and an image point of an optical system are said to be conjugate points. In terms of imaging, therefore, as is well recognized in the art, when an image of an object placed at the location of one of the optical conjugates is formed at the location of the second optical conjugate. A 4f optical system of an embodiment is configured to perform a cascade of two Fourier Transforms (FTs) with respect to light distribution at the entrance of the system. Generally, and for the purposes of this disclosure and invention as claimed, the two lens elements of the 4f optical system may have different focal lengths. In a specific case, the two lens elements of the 4f optical system have substantially equal focal lengths.

Example of a System.

[0020] According to the idea of the invention, the transmission (Tx) gratings are used at the input and output of the common-path interferometer for light-beam splitting/combining; the only, single arm of such interferometer (along which the reference and signal beams, formed by diffracted beams of +1 and -1 orders, propagate) is defined by as a lens system configured as a 4f system; phase-delay optical element(s) is/are inserted across one or both of the reference and signal beams inside the interferometer before these beams are recombined together at an output diffraction grating to form, at a chosen image plane, a light distribution representing the results of interference between the signal and reference beams. For the purposes of this disclosure and claims, and unless specified otherwise, the term common-path interferometer refers to and is defined as an interferometer in which both the reference beam and the signal beam propagate, between the input and output beam-splitting / combining components, through the same lens system disposed to perform a cascade of two Fourier Transforms on light distribution at the input beam-splitting component and to place the light distribution resulting from such cascade of spatial transformation at the output beam- splitting component. Being a common-path interferometric set-up, this is highly stable to any changes in the phase of light propagating therethrough and/or length of the interferometric arm, which facilitates an effective capture of precisely controlled OPD-scans over a large FOV. The FT of the interferogram representing such OPD-scan is further formed and captured by the optical detector to recover the spectrum for each pixel in the FOV.

[0021] This is schematically presented in the embodiment 200 of the imaging spectrometer of Fig.

2A, which utilizes the interferometer 210. The interferometer 210 is inserted in the path of the light beam 214 received from an optical system 220 (a microscope objective in one specific case) and the additional optical sub-system labeled as AUX. In one specific implementation, the light beam 214 is a collimated beam representing an optical plane wave (that is, an optical wave the wavefront of which is substantially plane) incident onto the interferometer 210 from the optical system 220 through the sub-system AUX. Within the interferometer, as discussed below, two first-order diffracted beams 230A, 230B are formed. The interferograms, defined by light from the two diffracted beams 230A, 230B at the final image plane 234 for each (x,y) location of the field of view, are recorded at the optical detector 238. The first-order diffracted beams 230A, 230B are created by an input (or first) diffraction grating 242 (also labeled as Gl and positioned at an intermediate image plane 246) from the incident light 214 and form signal and reference beams of the interferometer 210. The zeroth order of diffraction (just like any other diffracted beam besides first-order diffraction beams formed by the grating 242) is preferably blocked from propagating down the optical axis of the interferometer.

[0022] The diffracted beams are optically relayed to the second image plane 280 by a 4f system that includes first and second lens elements 254, 258, which lens elements have respectively-corresponding focal lengths fi and Ϊ2. It is appreciated that, in general, the values of fi and Ϊ2 may be equal or different from one another - this does not alter the principle of operation of the proposed system. Plane 262, in which the focal point of the lens element 254 and the focal point of the element 258 coincide on the optical axis, is a Fourier transform plane of the 4f optical system, in which an image of an object viewed with the microscope 220 is formed in light from the collimated beam 214. In other words, the Fourier transform plane of the 4f optical system is located between the lens elements forming the system at one respectively-corresponding focal length away from each of these lens element.

[0023] At the Fourier transform plane 262 of the 4f optical system formed by lens element 254, 258, an OPD is variably introduced between the beams 230A, 230B with the use of prismatic elements 266, 272 (or with the use of alternative means discussed below). Specifically, as discussed below, the OPD between the beams 230A, 230B can be continuously varied with the use of the prismatic elements 266, 272 or continuously or discretely varied with the use of the alternative means. After the variable OPD has been introduced, the beams 23 OA, 230B are further relayed by the lens 258 to the second (output) diffraction Tx- grating 276 (G2) disposed at the second image plane 280 to be recombined into combined light output 284. [0024] The combined light output 284 is relayed with a telescopic arrangement 288 formed by another 4f system that includes lens elements 290, 292 and that is complemented with a opaque screen, containing an aperture A and disposed to block the off-axis diffracted orders at the Fourier transform plane 294 of the arrangement 288. Again, the focal points of the lens elements 290, 292 coincide on axis at the Fourier transform plane 294 between the lens elements 290, 292.

[0025] The two on-axis components of the combined light output 284 interfere at the detector plane

234. The data-processing circuitry (not shown), operably connected to the detector 238 acquires output interferometric data from each of the detector pixels representing an interferogram to recover the

spectrum/spectra of source(s) of light at the optically-conjugate object plane viewed by the microscope 220.

[0026] In one specific example of the system 200,

transmission grating beamsplitters (Edmund Optics, Stock No. #46-073) are utilized as diffraction gratings 242, 276 that have groove-density of 110 Grooves/mm, and area of 12.7 mm 2 ; achromatic doublets with SM2-Threaded Mount and ARC of 400-700 nm (Thorlabs, AC508-150-A-ML) are used as lenses 254, 258 (fi =f 2 =150 mm, diameter of 2"); the achromatic doublet, SM2-Threaded Mount, ARC of 400-700 nm (Thorlabs, AC508-100-A-ML) is used as the lens 290 (f 3 =100 mm, diameter of 2"); and the achromatic doublet, SM2-Threaded Mount, ARC of 400-700 nm (Thorlabs, AC508-200-A-ML) is used as the lens 292 (f 4= 200 mm, diameter of 2"). PROSILICA GE 680 see

alliedvision.com/en/products/cameras/detail/680.html) is used as the optical detector 238.

[0027] Notably, in advantageous contradistinction with, for example, the imaging spectrometer utilizing the Sagnac interferometer, the present imaging spectrometer employing the common-path interferometer of Fig. 2A exhibits the phase of light that remains uniform across the whole FOV of the optical detector, i.e. identical for any point within such FOV. In other words, each pixel of the optical detector 238 receives the signal corresponding to the same value of the OPD between the reference and signal beams.

[0028] Fig. 2B provides a more detailed example itemizing the optical sub-system AUX of Fig. 2A.

Here, light from the optionally spectrally-tunable Laser is delivered through the optical system 220 with the use of the beamsplitter Dio to irradiate the object, and light emanating from the object is collected and directed towards the interferometer 500 with the reflector M, the collimating lens element L, and spectral filter F to form the input beam 214.

[0029] In further reference to Figs. 2A, 2B, the embodiment 200 may be optionally equipped with programmable electronic circuitry 294 (containing a programmable computer processor, for example) configured to govern the operation of the laser and/or determine data representing spectral distribution of light incident onto the detector 238 and/or determine data representing image(s) of the object, formed at the detector in different spectral bands, as well as, optionally. In one implementation, the electronic circuitry 294 is additionally programmed to operate, via appropriate electronic drives, the variable phase-delay element(s) as discussed below, i.e. scan the value representing the optical path difference between the reference and signal beams propagating through the common-path interferometer.

Embodiments of Variable Phase-Delay Element Configured for Scanning the Value of OPD.

[0030] Three independent techniques are discussed below to introduce an OPD between the reference and signal beams of the common-path interferometer of the system of the invention. These techniques can be used individually or in combination, depending on the end-user application.

[0031] Example 1: Prism-based OPD scanning.

[0032] The combination of prisms 300 discussed below is positioned across the reference and signal beams in the set-up 200 in plane 262. As schematically shown in Fig. 3, a translation scanning mechanism 310 is employed to carry a first prismatic element or prism 316 (made of optical material such as glass, for example, and having a wedge 316A at one end of it) and to repositionably move the wedge 316A of the prism 316 along an axis shown with an arrow 320 with respect to a second prismatic element or prism 324, the shape of which is complementary to the shape of the wedge 316. Phrased differently, the first prism 316 is formed to include a V-shaped wedge 316A extending outwardly at part of the body of the prism 316, while the second prism 324 is formed to include a V-groove 324A extending inwardly in the body of the prism 324. The shapes of the V-shaped wedge 316A and the V-groove 324A are complementary to one another. As shown, the prisms 316, 324 form a dove-tail type of contraption in which the protruding wedge of the prism 316 fits into the complementarily-shaped gap indentation of another.

[0033] The scanning mechanism 310, in operation, varies the thickness of the airgap 328 between the wedges 316, 324, the variation of which airgap in turn introduces a variable change in the optical path length (OPL) of the beam 23 OA as compared to that of the beam 230B. In reference to Fig. 2A, the fixed phase that beam 230B accrues upon transmission through the prism 324 is φ, while the phase φ + Δφ that beam 230A accrues upon transmission through the portions of the prisms 316, 324 and the gap 328 is varied due to variation of Δφ attributed to the instantaneous thickness of the airgap 328. It is understood, therefore, that, generally, the variable phase-delay optical element 300 includes first and second prisms having respectively-corresponding portions with first and second shapes that are complementary to one another. [0034] Example 2: OPD scanning due to rotation of an optically-isotropic transparent slab of material.

[0035] In an alternative embodiment 440 of Fig. 4, two thin optically-isotropic transparent slabs

444A, 444B of equal thickness are inserted, respectively, across +1 order and -1 order diffracted beams 230A, 230B (in place of wedges discussed in reference to embodiment 300), and one of the slabs - as shown, 444A - is caused to rotate to increase the OPL for one of the beams (as shown - beam 23 OA), for example with the use of a motor (not shown) operably cooperated with the slab. It is understood, therefore, that the variable phase-delay optical element of the common-path interferometer in this embodiment includes a slab of optical material, and the variable phase-delay optical element is configured to rotate such slab about an axis that is perpendicular to the axis of the 4f optical system.

[0036] The slabs can be made of glass or plastic, for example. It is appreciated that, if this approach is chosen in practice, the relative OPD between the beams 230A, 230B can be made zero and increased but not decreased, resulting in generation of only single-sided interferograms at the plane 234 of Fig. 2A. The OPD adjustment can be practically extended to allow for generation of a double-sided interferogram at plane 234 by independently rotating the slab 444B as well.

[0037] Example 3: Phase-plate-based OPD Stepping

[0038] In yet another alternative embodiment, a judiciously structured phase plate is used to achieve the user-defined variation of the OPD between the reference and signal beams of the interferometer of the embodiment. As shown in top and side plan views of Figs. 5 A, 5B, 5C an embodiment 500 of the phase plate includes a disk made of optically transparent material one surface of which (for example, back surface shown as 500-A) is flat, while another surface of which (for example, a front surface shown as 500-B) is shaped to define several steps the planar levels of which sequentially differ from one another by a predetermined nonzero height Ah. Such phase plate is referred to herein as a step-phase plate. As shown in the embodiment 500, one half of the step-phase plate (labeled as 500-R) has a thickness of AH, while the other half is divided into several segments (sectors, the angular measures of which as viewed from the axis of rotation 510 of the step-phase plate are equal to one another). The half of the step-phase plate having thickness AH at any point across such half is a reference portion of the step-phase plate (labeled as R). A sector that is immediately- adjacent to the reference portion (shown as sector 500-0) has the same thickness AH as the reference portion. The sector 500-1, which is immediately adjacent to the sector 500-0, has a thickness (AH + Ah). The sector 500-2, which is immediately adjacent to the sector 500-1, has athickness (AH+ 2Ah); and so on.

[0039] Generally, the number N of such immediately-adjacent to one another sectors is limited only by the width of a diffracted beam formed by the grating 242 (as shown in this example, by the cross-sectional width of the beam 23 OA): in operation, the footprint of the diffracted beam formed at the sector of the step- phase plate should not exceed the local width of the sector. As shown in embodiment 500, the step-phase plate includes six (6) sectors (labeled 500-0, 500-1, 500-2, 500-3, 500-4, and 500-5) in addition to the reference half 500-R of the step-phase plate. Each of these sectors subtends the same angle of 30 degrees as viewed fro the axis of rotation 510 of the plate 500.

[0040] Fig. 5C illustrates a cross-section of the step-phase plate 500 along the straight line 520, which connects the footprints of the diffracted beams 23 OA, 230B formed in operation of the system at the step-phase plate. As shown in the example of Fig. 5C, beams 230A, 230B are traversing, respectively, the sector 500-1 and the reference portion 500-R.

[0041] It is appreciated that, in operation, a step-phase plate (such as, for example, a six-sector step-phase plate shown in Figs. 5A, 5B, 5C) is inserted across the beams 230A, 230B at the Fourier transform plane 262 to from the variable phase-delay optical element configured to introduce a phase delay into one of the beams 230A, 230B with respect to another as a function of the angle of rotation of the step-phase plate about the axis 510. Generally, the variable phase-delay between the beams 23 OA, 230B will be decreased (by an increment equal to the phase acquired by light from the beam 23 OA upon propagation through a Δ 2-thick layer of the step-phase plate) each time the step-phase plate is rotated by an angle of (180/N) degrees. In the specific example of the plate 500, the variable phase-delay will be decreased (by an increment equal to the phase acquired by light from the beam 23 OA upon propagation through a Δ 2-thick layer of the plate 500) each time the plate 500 is rotated by an angle of 30 degrees about the axis 510 in the counterclock-wise direction. If the rotation of the step-phase plate is arranged in the clockwise direction then the variable phase- delay between the beams 230A, 230B (and, therefore, the optical path difference between these two beams) is, accordingly, incrementally increased. It is appreciated, therefore, that in one embodiment the variable phase-delay optical element of the interferometer includes a step-phase plate of optical material, and that such variable phase-delay optical element is configured to rotate the step-phase plate about an axis of the plate that is parallel to the axis of the 4f optical system.

[0042] Double sided interferograms can be generated by stepping the phase plate up to 360 degrees with respective step angles.

[0043] To increase the OPD scanning resolution or scanning range, multiple step-phase plates can be used in combination as shown in the example of Fig. 6. Here, three step-phase plates 600A, 600B, 600C are shown to be positioned co-axially, one after another across the beams 230A, 230B (as a stack of closely- disposed step-phase plates) at the Fourier transform plane 262. The increments of change of the OPD, defined by at least two of the plates 600A, 600B, 600C are different from one another. In the example of Fig. 6, such increments are different for all the plates and are defined by layers of step-phase plate material having thicknesses Ahl, Ah2, and Ah3, respectively. Optionally, each of the plates 600A, 600B, 600C is configured to be rotated independently about the axis 510. It is appreciated, therefore, that the variable phase-delay optical element of the interferometer may include a stack of optical step-phase plates at least two of which define unequal increments of the phase delay to be introduced between the signal and reference beams of the interferometer.

Principle of Operation of an Embodiment

[0044] In reference to Fig.2 A, and assuming the light distribution (optical signal) produced by the microscope 220 at the plane 246 to a plane-wave distribution,

[0045] E 0 (x, y, z, t) =∑ k E x, y, k)e ik t z~c (1),

[0046] the electric field immediately after the propagation through first grating 242 (that is, immediately to the right of it as shown in Fig.2A) can be expressed as

[0047] E 1 x, y, z, t) = E 0 x, y,z,t)x^x {e ik ° x + e ~ik ° x ) (2)

[0048] Then at the first Fourier plane 262, a finite phase delay, φ, is introduced to beam 230A as compared to beam 230B so that right before the second grating 276 the electric field can be expressed as

[0049] E 2 (x,y,z,t) = E 0 (x,y,z,t)x^x(e ik ° x + e ~ ^ k ° x -^) (3)

[0050] So adding the phase delay φ is equivalent to inserting in beam 230A a parallel glass slab of thickness T chosen such that

[0051] φ = kT(n glass - n air ) = kTAn (4),

[0052] where n glass and n^ are refractive indices of glass and air, respectively. Upon further propagation and after the second grating 276 the electric field can be expressed as :

[0053] E 3 x, y, z, t) = E 2 x, y,z,t)x x {e ik ° x + e ~ik ° x ) (5 A)

[0054] or

[0055] E 3 x, y, z, t) = E 0 x, y, z, t) X X {e i2k ° x + e -^ χ -φ) + 1 + e ίφ) ( 5B )

[0056] At the second Fourier plane 294 the off-axis diffraction orders are blocked from propagating further, and the electric field right before the plane 234 of the detector 238 is

[0057] E 4 (x,y,z, t) =∑ k E(x,y,k e ik ^- ct -'f ) /^ xjx (e^/ 2 + e " ^ 2 ) (6)

[0058] The intensity at the plane of the detector, therefore, is:

[0059] I det (x,y,O =∑ k E 2 (x,y,k)cos 2 g) = ∑ k E 2 (x,y, k)[l + cos (kTAn)] (7)

[0060] Such intensity, acquired by the detector 238, is a function of thickness T and represents the interferogram formed at the position (x,y) at plane of the detector. The first term in Eq. (7) is a DC term, while the second term contains the spectral information of light arriving at the detector from the optically- conjugate point at the sample (viewed under the microscope 220). As illustrated in Fig. 7, the intensity of the source spectrum \E 2 (x, y, k) | can be recovered by taking the cosine transform of the AC term of the interferogram of Eq. (7). Alternatively, the recovery of spectral information from the interferogram can be effectuated with the use of, for example, data-fitting techniques such as a fast orthogonal search (FOS) (see, for example, Korenberg, M J et al, , "Raman spectral estimation via fast orthogonal search." Analyst 122.9 (1997): 879-882; incorporated herein by reference)

[0061] As was already alluded to above, non-common path interferometers such as the conventional

Michelson interferometer, for example, are unstable with respect to phase disturbances. In contradistinction, an embodiment of the invention facilitates the measurement of interferograms caused by weak incoherent signals that the Michelson-based design, for example, cannot handle. To illustrate the degree of phase- stability of the new imaging FTS system structured around the novel common-path interferometer-based system 200, measurements of laser light reflected from a flat surface at the sample plane have been performed, with results presented in Figs. 8A, 8B, 8C, and 8D. Here, Fig. 8A presents a mean-intensity image based on measurements acquired at 1,000 time points over 50 seconds. Fig. 8B is a mean-phase image drawn from the same data set as that of Fig. 8A. Fig. 8C is a plot of phase-change with time occurring at the point PI in the image field of Fig. 8C. Fig. 8D is a plot of phase-change with time occurring at the point P2 in the image field of Fig. 8C. Insets labelled "AC*" in Figs. 8C, 8D provide plots representing respectively- corresponding autocorrelation of the change in phase angle as a function of time.

[0062] Similarly, in advantageous contradistinction with the Sagnac-interferometer-based FTS (in which the operation of the system is known to be substantially slowed down due to necessity to recover the zero-level OPD data, from the generally linearly-varied across the FOV, by scanning across the FOV in order to generate sufficient number of interferogram sample for all pixels in the FOV of the system), the operation of the present embodiment allows for random data acquisition and is not limited to sequential data acquisition like the Sagnac-based system. As will be readily appreciated by a skilled artisan, the Sagnac-based system is not effective to be used under conditions of random data-acquisition. Accordingly, unlike the Sagnac-based FTS, embodiments of the present invention are compatible and can be operated with high-throughput data- fitting algorithms such as FOS (which requires random data acquisition), while at the same time maintaining a near-uniform distribution of OPD across the FOV. A skilled artisan will readily recognize the latter from empirically-obtained information shown in Figs. 9A, 9B, 9C, 9D, and 9E, in which Figs. 9A, 9B, and 9C illustrate phase uniformity (observed via OPD scanning) across the FOV patterns in a Michelson

interferometer, in the embodiment of the common-path interferometer configured according to the idea of the invention, and in a Sagnac interferometer. Fig. 9D provides an image of a surface of a standard USAF target, formed in laser light reflected from such surface and acquired with the embodiment of system 200, while Fig. 9E shown the image of a specimen of pre-determined cells empirically acquired with a Sagnac-interferometer based spectrometer.

[0063] A skilled artisan will readily appreciate, therefore, that implementations of the idea of an imaging spectrometer based on the proposed common-path interferometer lend themselves to be an add-on to any commercially-available widefield fluorescence microscope system, thereby enabling high-throughput hyperspectral imaging capabilities to the already-existing microscopes. Along with custom-designed data- processing algorithms, the proposed FTS system can be used to uniquely separate close emission spectra from known biological fluorophores. The proposed design can also be used with most of the high throughput imaging cytometer systems to enhance their information manifold. Furthermore, the embodiments of the spectrometer in combination with structured light illumination can be used to design new depth resolved wide field hyperspectral imaging systems. Furthermore, the proposed interferometer can also be used to do faster Raman imaging.

[0064] In accordance with examples of embodiments, a unique common-path interferometric system and an imaging spectrometer employing such common-path interferometric system are provided. While the disclosure recites specific values chosen for these embodiments are recited, it is to be understood that, within the scope of the invention, the values of any or all of parameters may vary over wide ranges to suit different applications. It will be understood by those of ordinary skill in the art that modifications to, and variations of, the illustrated embodiments may be made without departing from the inventive concepts disclosed herein. For example, any of the diffraction gratings 242, 276 of the embodiment 200 (discussed in reference to Fig. 2A) can be configured to operate in reflection, in which case the corresponding embodiment of the common-path interferometer (not shown) will have a folded optical path for light propagating therethrough.

[0065] Alternatively, a related embodiment 1000 of the common-path interferometer and an imaging spectrometer employing such common-path interferometer, as shown schematically in Fig. 10, may be used. In comparison with the embodiment 200 of Fig. 2A, the embodiment 1000 includes only one diffraction grating 242 disposed at a distance f^ in front of the 4f optical system comprising the lens elements 254, 258, and instead of the grating 276 present at the second intermediate image plane 280, the optical detector 238 is disposed at the plane 280 to directly acquire the interferogram formed by the beams 230A, 230B that have propagated through the 4f optical system. While the variable phase-delay optical element 1010, positioned at the Fourier transform plane 262 within the 4f optical system, is shown as a combination of prisms discussed in reference to embodiments 200 and 300, it is appreciated that any of the embodiments of the variable phase-delay optical elements of Figs. 4, 5A, 5B, 5C, and Fig. 6 can be used instead. An imaging spectrometer 1000 has a phase distribution of light, received by the optical detector 238, that is sinusoidally-modulated across a field of view (FOV) defined by light that has propagated from the microscope through the 4f imaging system towards the detector. Just as in the embodiment 200, the embodiment 1000 may be equipped with programmable electronic circuitry (not shown) configured to govern the operation of the laser and/or determine data representing spectral distribution of light incident onto the detector 238 and/or data representing image(s) of the object, formed at the detector in different spectral bands.

[0066] The present invention also provides a method for performing imaging spectroscopy, in which light from a collimated beam of light, delivered with a microscope from an object, is propagated through a common-path interferometer including a 4f optical system; the light transmitted through the common-path interferometer is further acquired with an optical detector to determine spectral distribution of said light.

The process of imaging spectroscopy according to the embodiment of the invention additionally includes defining a distribution of phase of collimated light that has propagated through such common-path interferometer, to be sinusoidally-modulated across a field of view (FOV) of the optical detector.

Alternatively or in addition, the step of propagating includes (i) forming first and second diffracted beams from the collimated beam of light transmitted through a first diffraction grating disposed in a first plane; and (ii) propagating the first and second diffracted beams through the 4f optical system to transmit light from the first and second diffracted beams through a second diffraction grating disposed in a second plane, wherein the second plane is optically-conjugate to the first plane in light propagated through the 4f optical system. In a specific embodiment, the method may further include transmitting light from a collimated beam of light, that has propagated through the 4f optical system, through an auxiliary 4f optical system positioned co-axially with the 4f optical system to form an interferogram at the optical detector, when the optical detector is disposed at a plane that is optically-conjugate to the first plane. In such a specific case, the distribution of phase of light acquired by so-positioned optical detector, is defined to be uniform across a field of view of the optical detector. The method may further include a step of optically delaying one of the first and second diffracted beams with a spatially-repositionable variable phase-delay optical element disposed in a Fourier transform plane, which Fourier transform plane is defined by a lens element of the 4f optical system with respect to the collimated beam of light delivered by the microscope from the object. In related embodiments, the step of optically delaying may include (a) disposing first and second optical prisms across one of the first and second diffracted beams, where such first and second optical prisms have respectively-corresponding portions with first and second shapes that are complementary to one another; or (b) disposing a slab of optical material across one of the first and second diffracted beams, where the variable phase-delay optical element is configured to rotate such slab about an axis that is perpendicular to the axis of the 4f optical system; or (c) positioning a step-phase plate of optical material across one of the first and second diffracted beams, where the variable phase-delay optical element is additionally configured to rotate said step-phase plate about an axis of said plate, the axis of said plate being parallel to the axis of the 4f optical system.

Methodology of Compressive Spectral Recovery

[0067] Compressive Spectral Recovery methodology, used with an embodiment of the invention to procure the data from the optical interferogram acquired by the detector 238, is further discussed below.

[0068] The FTS measurement corresponding to each value of the OPD (i.e. I det (T)) provides a weighted sampling of the full spectral range (i.e. E 2 (: )), not an individual spectral band.

[0069] I det (x, y, T) = ∑ k E 2 (x, y, k) [1 + cos (kTAn)] (8)

[0070] Hence, each FTS measurement provides information about all wavelengths in the spectrum; therefore, it may be amendable in some cases to extract full spectral information with sparse sampling especially when prior information is available or when only partial statistics of the spectrum (such as peak emission wavelengths) are of interest. For example, compressive algorithms, such as fast orthogonal search (FOS), can be used with significantly less number of measurements than exhaustive sampling of Fourier space (see, for example, Korenberg et al., "Raman spectral estimation via fast orthogonal search." Analyst 122.9 (1997): 879-882.)

[0071] The results of the simulations shown in Figs. 11 A, 11B, 11C, 11D suggest that imaging FTS methodology of the present invention can compressively measure the peak emission wavelength of at least a quantum dot in a 400 nm spectral range with an accuracy of lnm and with as few as 7 measurements in noiseless conditions (see Fig. 1 IC). For comparison, wavelength-tunable-filter-based spectrometers require at least 200 measurements in similar conditions. Furthermore, in the presence of noise the same results can be achieved with less than 2000 photons and 20 measurements using the imaging FTS methodology of the present invention (see Fig. 1 ID), while tunable-filters-based spectrometers cannot detect at least a single photon per band with same number of photons due to its poor optical throughput.

[0072] It is appreciated, however, that in order to achieve such high compression levels with the use of the imaging FTS of the present invention the OPD positions (at which the measurements are taken) should preferably be optimized. Fig. 1 IC illustrates the measurement error, defined as a difference between the measured peak wavelength and the actual peak wavelength, i.e.,

\measured peakwavelength - actual peak wavelength\, for the same number of OPD samples when the samples are taken at random. Notably, some sample combinations performs extremely well than the average case. For instance, for 6 OPD positions the best set gives near zero error while the measurement error is more than 200 nm on average. It is preferred, therefore, that the measurements are taken on these best OPD positions referred to hereinafter as as "optimized OPD positions", OOPs). The process of measurement of partial interferogram only at these optimized OPD positions is referred to as "optimized compressed measurements", or OCMs.

[0073] Since the measurements are preferably taken at the optimized OPD positions (for all the pixels in the FOV), it may be important that all the pixels of the FOV correspond to the same OPD once the interferometer steps in to an optimized OPD position. In other words, the OPD (or, alternatively, the phase of light) should be uniform across the FOV in order to perform optimized compressed measurements. Sagnac- interferometer based system is not capable of performing optimized compressed measurements simply because (a) it inevitably has a phase tilt across the FOV and therefore (b) it cannot "step" in to optimized OPD positions for all the pixels in the FOV at the same time. Therefore, attempting to perform optimized compressed measurements with Sagnac is highly inefficient.

[0074] Finding the OOPs is not a straightforward task. According to the present methodology, full interferograms are measured for known light sources that illuminate the entire FOV at a time and then random OPD sets are chosen to recover the wavelength maximum (wavelength corresponding to the peal, or peal wavelength) to find out OOPs in an exhaustive search. Once OOPs are found, they can be recorded and OCMs can be performed for the specimens of interest. The procedure is summarized below.

[0075] Example.

[0076] The optimum OPD positions were selected as following. Emission light from Green LED source (Thorlabs, M530L3) and Red LED source (Thorlabs, M625L3) were measured throughout the entire FOV with Nyquist sampling criterion. The spectral cubes were recovered using Fourier transformation procedure and ground truth peak-wavelengths were extracted. Then for a specified compression ratio n/N (where n = number of compressive measurements, N= number of Nyquist measurements), n measurements were drawn at random OPD positions. These were then used to recover the peak wavelength using a FOS like algorithm. Then the measurement error, i.e. absolute difference of recovered peak from the ground truth, was calculated. This was repeated a sufficiently large number of times and the OPD position set which resulted the minimum measurement error for both green and red LEDs was chosen as the OOPs.

[0077] Following the above steps, the interference images for test specimens were taken at OOPs and the peak wavelengths were recovered using FOS. Figs. 12A , 12B, 12C, 12D, 12E, and 12F show the peak wavelengths respectively for no compression (i.e. compression ratio of n/N =1, Figs. 12A, 12E), compression ratio of n/N=0.2 (Figs. 12B, 12E), and compression ratio of n/N= 0.05 (Figs. 12C, 12F) for the fluorescent bead specimen. Fig. 13A, 13B, 13C, 13D, 13E, and 13F show results of the same imaging procedure performed with the mouse muscle specimen. For beads acceptable results were acquired up to lowest compression ratio of 0.05, while for mouse muscle specimen the scheme broke down after 0.2 (compare Figs. 13E and 13F). In Figs. 14A, 14B, we quantitatively compare our optimized sampling strategy with conventional random sampling in compressive sensing. Agreeing with the qualitative results, for the mouse muscle specimen, average measurement error of optimized sampling reached the best case of random sampling with n/N = 0.2. For that compression ratio measurement error for average case in random sampling remained relatively higher and the worst case was unacceptably high. The average and worst case of random sampling seemed to converge after n/N=0.5 . This suggests that the proposed strategic sampling scheme guarantees near best results for compression ratios above a certain threshold (0.2 for mouse muscle specimen)

[0078] It is appreciated, therefore, that embodiments of the methodology of compressive spectral recovery provide a method for measuring of a spectral characteristic at all pixels of a two-dimensional scene. Such method includes: for each of a plurality of chosen light sources, forming a respectively- corresponding interferogram at an output of an imaging spectrometer (where the imaging spectrometer is configured to produce, at the output, a distribution of light having uniform distribution of phase across a field-of-view (FOV) corresponding to said output, and where each of the plurality of chosen light source has a known spectral signature). The method further includes randomly sampling each of formed interferograms with varying number of samples and at optical- path-difference (OPD) values to obtain data; and fitting said data to respectively-corresponding known spectral signatures of the plurality of chosen light source to calculate values representing accuracies of said fitting. The OPD values represent values of OPD of light propagating through the imaging spectrometer. The method additionally includes selecting OPD values corresponding to the smallest number of samples that satisfy a requirement of a threshold accuracy of fitting; and performing interference measurements of the two-dimensional scene at each of selected OPD positions to recover the spectral characteristic. The method may additionally include non- randomly varying the OPD values, which representing values of OPD between reference and signal beams of an interferometer of the imaging spectrometer. Alternatively or in addition, the method may include the steps of transmitting light through a common-path interferometer of the imaging spectrometer and/or non-randomly changing the values of OPD between reference and signal beams of such common-path interferometer.

[0079] For the purposes of this disclosure and the appended claims, the use of the terms

"substantially", "approximately", "about" and similar terms in reference to a descriptor of a value, element, property or characteristic at hand is intended to emphasize that the value, element, property, or characteristic referred to, while not necessarily being exactly as stated, would nevertheless be considered, for practical purposes, as stated by a person of skill in the art. These terms, as applied to a specified characteristic or quality descriptor means "mostly", "mainly", "considerably", "by and large", "essentially", "to great or significant extent", "largely but not necessarily wholly the same" such as to reasonably denote language of approximation and describe the specified characteristic or descriptor so that its scope would be understood by a person of ordinary skill in the art. In one specific case, the terms "approximately", "substantially", and "about", when used in reference to a numerical value, represent a range of plus or minus 20% with respect to the specified value, more preferably plus or minus 10%, even more preferably plus or minus 5%, most preferably plus or minus 2% with respect to the specified value. As a non-limiting example, two values being "substantially equal" to one another implies that the difference between the two values may be within the range of +/- 20% of the value itself, preferably within the +/- 10% range of the value itself, more preferably within the range of +/- 5% of the value itself, and even more preferably within the range of +/- 2% or less of the value itself.

[0080] The use of these term in describing a chosen characteristic or concept neither implies nor provides any basis for indefiniteness and for adding a numerical limitation to the specified characteristic or descriptor. As understood by a skilled artisan, the practical deviation of the exact value or characteristic of such value, element, or property from that stated falls and may vary within a numerical range defined by an experimental measurement error that is typical when using a measurement method accepted in the art for such purposes.

[0081] For example, a reference to an identified vector or line or plane being substantially parallel to a referenced line or plane is to be construed as such a vector or line or plane that is the same as or very close to that of the referenced line or plane (with angular deviations from the referenced line or plane that are considered to be practically typical in related art, for example between zero and fifteen degrees, preferably between zero and ten degrees, more preferably between zero and 5 degrees, even more preferably between zero and 2 degrees, and most preferably between zero and 1 degree). For example, a reference to an identified vector or line or plane being substantially perpendicular to a referenced line or plane is to be construed as such a vector or line or plane the normal to the surface of which lies at or very close to the referenced line or plane (with angular deviations from the referenced line or plane that are considered to be practically typical in related art, for example between zero and fifteen degrees, preferably between zero and ten degrees, more preferably between zero and 5 degrees, even more preferably between zero and 2 degrees, and most preferably between zero and 1 degree). A term "substantially rigid", when used in reference to a housing or structural element providing mechanical support for a contraption in question, generally identifies the structural element that rigidity of which is higher than that of the contraption that such structural element supports. As another example, the use of the term "substantially flat" in reference to the specified surface implies that such surface may possess a degree of non-flatness and/or roughness that is sized and expressed as commonly understood by a skilled artisan in the specific situation at hand. [0082] Embodiments of the invention have been described as including a processor or electronic circuitry controlled by instructions stored in a memory. Those skilled in the art should also readily appreciate that instructions or programs defining the functions of the present invention may be delivered to a processor in many forms, including, but not limited to, information permanently stored on non-writable storage media (e.g. read-only memory devices within a computer, such as ROM, or devices readable by a computer I/O attachment, such as CD-ROM or DVD disks), information alterably stored on writable storage media (e.g. floppy disks, removable flash memory and hard drives) or information conveyed to a computer through communication media, including wired or wireless computer networks. In addition, while the invention may be embodied in software, the functions necessary to implement the invention may optionally or alternatively be embodied in part or in whole using firmware and/or hardware components, such as combinatorial logic, Application Specific Integrated Circuits (ASICs), Field-Programmable Gate Arrays (FPGAs) or other hardware or some combination of hardware, software and/or firmware components.

[0083] References throughout this specification to "one embodiment," "an embodiment," "a related embodiment," or similar language mean that a particular feature, structure, or characteristic described in connection with the referred to "embodiment" is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment," "in an embodiment," and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment. It is to be understood that no portion of disclosure, taken on its own and in possible connection with a figure, is intended to provide a complete description of all features of the invention.

[0084] Within this specification, embodiments have been described in a way that enables a clear and concise specification to be written, but it is intended and will be appreciated that embodiments may be variously combined or separated without parting from the scope of the invention. In particular, it will be appreciated that all features described herein at applicable to all aspects of the invention.

[0085] In addition, when the present disclosure describes features of the invention with reference to corresponding drawings (in which like numbers represent the same or similar elements, wherever possible), the depicted structural elements are generally not to scale, and certain components are enlarged relative to the other components for purposes of emphasis and understanding. It is to be understood that no single drawing is intended to support a complete description of all features of the invention. In other words, a given drawing is generally descriptive of only some, and generally not all, features of the invention. A given drawing and an associated portion of the disclosure containing a description referencing such drawing do not, generally, contain all elements of a particular view or all features that can be presented is this view, at least for purposes of simplifying the given drawing and discussion, and directing the discussion to particular elements that are featured in this drawing. [0086] A skilled artisan will recognize that the invention may possibly be practiced without one or more of the specific features, elements, components, structures, details, or characteristics, or with the use of other methods, components, materials, and so forth. For example, the interferometer system such as the system 200 of Fig. 2A can be appropriately modified such that diffracted beams of different orders - in one instance, the zeroth and +lst orders of diffraction - are chosen as the reference and signal beams propagating between the gratings Gl, G2, while other diffraction orders generated by the grating Gl are blocked from propagation towards G2. Generally, therefore, the reference and signal beams are chosen from the first and second diffracted beams formed by the diffraction grating Gl and inclined at different angles with respect to the axis of the common-path interferometer. In the specific case of Fig . 2A, where the + 1 st and - 1 st diffracted order beams are used as reference and signal beams, one of these beams is inclined at a negative first angle with respect to the axis of the interferometer, while another is inclined at a positive (and therefore different from the fist angle) angle with respect to the axis of the interferometer.

[0087] Therefore, although a particular detail of an embodiment of the invention may not be necessarily shown in each and every drawing describing such embodiment, the presence of this particular detail in the drawing may be implied unless the context of the description requires otherwise. In other instances, well known structures, details, materials, or operations may be not shown in a given drawing or described in detail to avoid obscuring aspects of an embodiment of the invention that are being discussed. Furthermore, the described single features, structures, or characteristics of the invention may be combined in any suitable manner in one or more further embodiments. Disclosed aspects, or portions of these aspects, may be combined in ways not listed above. Accordingly, the invention should not be viewed as being limited to the disclosed embodiment(s).