WO2023145207A1 - 分光測定装置、及び分光測定方法 - Google Patents

分光測定装置、及び分光測定方法 Download PDF

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Publication number
WO2023145207A1
WO2023145207A1 PCT/JP2022/042752 JP2022042752W WO2023145207A1 WO 2023145207 A1 WO2023145207 A1 WO 2023145207A1 JP 2022042752 W JP2022042752 W JP 2022042752W WO 2023145207 A1 WO2023145207 A1 WO 2023145207A1
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fibers
sample
light
fiber unit
face
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English (en)
French (fr)
Japanese (ja)
Inventor
康昭 熊本
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University of Osaka NUC
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Osaka University NUC
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Priority to JP2023576643A priority Critical patent/JP7782868B2/ja
Priority to US18/729,988 priority patent/US20250116552A1/en
Priority to EP22924059.3A priority patent/EP4471392A4/en
Publication of WO2023145207A1 publication Critical patent/WO2023145207A1/ja
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/0218—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using optical fibers
    • G01J3/0221—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using optical fibers the fibers defining an entry slit
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J1/00—Photometry, e.g. photographic exposure meter
    • G01J1/02—Details
    • G01J1/04—Optical or mechanical part supplementary adjustable parts
    • G01J1/0407—Optical elements not provided otherwise, e.g. manifolds, windows, holograms, gratings
    • G01J1/0437—Optical elements not provided otherwise, e.g. manifolds, windows, holograms, gratings using masks, aperture plates, spatial light modulators, spatial filters, e.g. reflective filters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/021—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using plane or convex mirrors, parallel phase plates, or particular reflectors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/0218—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using optical fibers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/0229—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using masks, aperture plates, spatial light modulators or spatial filters, e.g. reflective filters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0205—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows
    • G01J3/0248—Optical elements not provided otherwise, e.g. optical manifolds, diffusers, windows using a sighting port, e.g. camera or human eye
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/027—Control of working procedures of a spectrometer; Failure detection; Bandwidth calculation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/0294—Multi-channel spectroscopy
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/02—Details
    • G01J3/10—Arrangements of light sources specially adapted for spectrometry or colorimetry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/12—Generating the spectrum; Monochromators
    • G01J3/18—Generating the spectrum; Monochromators using diffraction elements, e.g. grating
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/28—Investigating the spectrum
    • G01J3/44—Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/65—Raman scattering

Definitions

  • the present invention relates to a spectroscopic measurement device and a spectroscopic measurement method, and more particularly to a spectroscopic measurement device and a spectroscopic measurement method that spectroscopically measure signal light such as Raman scattered light generated in a sample to generate a spectroscopic image.
  • Patent Document 1 and Non-Patent Documents 1 to 3 disclose methods for measuring Raman spectra.
  • Patent Document 1 discloses a multifocal confocal Raman microscope using a fiber bundle. Light from the pinhole array is incident on the incident end of the fiber bundle.
  • a fiber bundle is arranged in front of the spectroscope.
  • fibers are arranged in a row at the output end.
  • Non-Patent Document 2 multifocus is generated by modulating illumination light with a liquid crystal spatial light modulator.
  • Non-Patent Document 3 discloses a multifocal confocal Raman spectroscopy microscope.
  • Non-Patent Document 3 uses a microlens array to generate multifocus.
  • a multi-fiber is used.
  • the measurement time becomes long.
  • imaging takes a long time because a plurality of spatial coordinate positions are sequentially measured. If the measurement time per coordinate is shortened, the effect of noise will occur. Therefore, there is a demand to measure an appropriate spectral image in a short time.
  • the present disclosure has been made in view of the above points, and an object thereof is to provide a spectroscopic measurement apparatus and spectroscopic measurement method capable of generating an appropriate spectroscopic image with short-time measurement.
  • a spectrometer includes a light source that generates illumination light, a spectroscope that separates signal light from a sample illuminated by the illumination light and detects it with a two-dimensional array photodetector, the sample to the spectroscope, wherein the plurality of fibers are arranged adjacent to each other on the input end face of the fiber unit, and the plurality of fibers are arranged on the output end face of the fiber unit.
  • the detection result of the two-dimensional array photodetector with reference to the fiber units arranged in a multi-line pattern with a gap and the arrangement relationship of the plurality of fibers on the incident end surface and the output end surface of the fiber unit.
  • a processing unit that generates a spectroscopic image of the sample from.
  • the above spectroscopic measurement apparatus further includes a spatial light modulator that modulates the illumination light from the light source so as to selectively illuminate a plurality of points on the sample, and the signal light from the plurality of points passes through the fiber unit. It may be made to each enter into either of the said fibers.
  • the above spectroscopic measurement apparatus may further include a camera that captures an optical image of the sample, and selectively illuminate a plurality of points on the sample that are extracted based on the optical image.
  • the plurality of fibers may be arranged in a hexagonal close-packed arrangement on the incident end surface of the fiber unit.
  • the incident end face of the fiber unit may be arranged at a position conjugate with the sample.
  • the signal lights emitted from the different fibers may be detected so as not to overlap on the light receiving surface of the two-dimensional array photodetector.
  • the spectroscopic measurement method comprises the steps of illuminating a sample with illumination light from a light source, and transmitting signal light from the sample to a fiber unit from an incident end surface in which a plurality of fibers are arranged adjacently. emitting from the output end face of the fiber unit in which the plurality of fibers are arranged in a multi-line pattern with intervals; splitting the signal light emitted from the output end face into two-dimensional A spectroscopic image of the sample from the detection result of the two-dimensional array photodetector, with reference to the step of detecting with an array photodetector and the arrangement relationship of the plurality of fibers on the incident end face and the exit end face of the fiber unit. and generating
  • the spatial light modulator modulates the illumination light from the light source so as to selectively illuminate a plurality of points on the sample, and the signal light from the plurality of points is transmitted to the fiber of the fiber unit. You may make it each inject into either.
  • an optical image of the sample may be captured by a camera, and a plurality of points on the sample extracted based on the optical image may be selectively illuminated.
  • the plurality of fibers may be arranged in a hexagonal close-packed arrangement on the incident end surface of the fiber unit.
  • the incident end surface of the fiber unit may be arranged at a position conjugated to the sample.
  • the signal lights emitted from the different fibers may be detected so as not to overlap on the light receiving surface of the two-dimensional array photodetector.
  • a spectroscopic measurement device and spectroscopic measurement method capable of generating an appropriate spectroscopic image in a short time measurement.
  • FIG. 1 is a schematic diagram showing a spectrometer according to Embodiment 1.
  • FIG. It is a figure which shows fiber arrangement
  • FIG. 4 is a diagram for explaining extraction processing of a ROI (Region Of Interest); It is a figure for demonstrating the spectroscopic measurement of the signal light from ROI.
  • 4A and 4B are diagrams showing images of the incident end and the exit end of the fiber unit 40.
  • FIG. 4 is a diagram showing spectroscopic measurement results of Example 1.
  • FIG. FIG. 10 is a diagram showing spectroscopic measurement results of Example 2;
  • FIG. 10 is a diagram showing spectroscopic measurement results of Example 3;
  • FIG. 10 is a diagram showing spectroscopic measurement results of Example 4;
  • FIG. 1 is a schematic diagram showing the optical system of the spectrometer 1.
  • the spectroscopic measurement device 1 is a spectroscopic microscope that captures a spectroscopic image of the sample S.
  • the spectrometer 1 measures spectral data of Raman scattered light from the sample S.
  • FIG. 1 is a schematic diagram showing the optical system of the spectrometer 1.
  • the spectroscopic measurement device 1 is a spectroscopic microscope that captures a spectroscopic image of the sample S.
  • the spectrometer 1 measures spectral data of Raman scattered light from the sample S.
  • the spectroscopic measurement apparatus 1 includes a spectral illumination optical system 10, an observation illumination light source 18, an observation optical system 20, a spectroscopic measurement optical system 30, a fiber unit 40, a spectroscope 50, and a processing device 60.
  • the spectral illumination optical system 10 is an optical system for guiding the illumination light L1 to the sample S. As shown in FIG.
  • the spectral illumination optical system 10 includes a light source 11 , a spatial light modulator 12 , a dichroic mirror 13 , a dichroic mirror 14 and a lens 15 .
  • the light source 11 is a laser light source that generates illumination light for spectrometry. Illumination light for spectroscopic measurement is simply referred to as illumination light L1.
  • the light source 11 is a DPSS (Diode Pumped Solid State) laser that emits CW (Continuous Wave) light with a wavelength of 660 nm.
  • DPSS Dynamic Switched Solid State
  • CW Continuous Wave
  • the type of light source 11 and the laser wavelength are not particularly limited.
  • the illumination light L1 serves as excitation light that excites the sample S. As shown in FIG. Therefore, the light source 11 generates monochromatic illumination light L1.
  • Illumination light L1 from the light source 11 enters the spatial light modulator 12 .
  • the spatial light modulator 12 spatially modulates the illumination light L1 based on the control signal from the processing device 60 .
  • the spatial light modulator 12 controls the spatial distribution of the illumination light L1 on the sample S. Thereby, on the sample S, the region on which the illumination light L1 is incident can be made into a desired shape and size. For example, the spatial light modulator 12 modulates the illumination light L1 so as to illuminate only the ROI of the sample.
  • the spatial light modulator 12 is, for example, a liquid crystal device such as LCOS (Liquid crystal on silicon).
  • the spatial light modulator 12 is a liquid crystal panel with pixels arranged in an array. By controlling the voltage applied to each pixel, the phase of the reflected light can be modulated.
  • the spatial light modulator 12 is not limited to a reflective liquid crystal device such as LCOS, and may be a transmissive liquid crystal device.
  • the spatial light modulator 12 is not limited to a liquid crystal device, and a DMD or the like can be used.
  • the illumination light L1 from the spatial light modulator 12 is incident on the dichroic mirror 13.
  • the dichroic mirror 13 splits light according to wavelength.
  • the dichroic mirror 13 reflects the wavelength of the illumination light L1. Therefore, the dichroic mirror 13 reflects the illumination light L1 toward the dichroic mirror 14 .
  • the dichroic mirror 14 splits light according to wavelength.
  • the dichroic mirror 14 transmits the wavelength of the illumination light L1. Therefore, the illumination light L ⁇ b>1 transmitted through the dichroic mirror 14 enters the lens 15 .
  • the lens 15 is an objective lens and converges the illumination light L1 on the sample S. Thereby, the sample S is illuminated with the illumination light L1. Further, the illumination light L1 is modulated by the spatial light modulator 12. FIG. Therefore, the illumination light L1 can illuminate a desired area on the sample S.
  • the sample S is mounted on a stage (not shown) or the like. In order to change the illumination position of the sample S, the stage may be a drive stage.
  • the observation illumination light source 18 is, for example, a lamp light source, and generates white observation illumination light L4.
  • the observation illumination light L4 from the observation illumination light source 18 illuminates the sample S.
  • the observation illumination light source 18 is not limited to a white lamp light source.
  • a non-monochromatic light source can be used for the observation illumination light source 18 .
  • the observation optical system 20 is an optical system for guiding the observation light L2 from the sample S to the camera 23.
  • the observation light L2 is light from the area illuminated by the observation illumination light L4.
  • the observation light L2 is scattered light scattered by the sample S, reflected light reflected by the sample S, fluorescence generated by the sample S, or the like.
  • the observation optical system 20 has a lens 15 , a dichroic mirror 14 , a dichroic mirror 13 , a filter 21 and a lens 22 .
  • the observation light L2 from the sample S is refracted by the lens 15 and enters the dichroic mirror 14.
  • the observation light L2 transmitted through the dichroic mirror 14 is incident on the dichroic mirror 13 .
  • the observation light L2 transmitted through the dichroic mirror 13 enters the filter 21 .
  • the filter 21 is an optical filter and transmits part of the light from the sample S.
  • the filter 21 is a wavelength filter that transmits or blocks light depending on the wavelength.
  • the camera 23, which will be described later can detect only the observation light L2 of the desired wavelength. In other words, light having a wavelength that passes through the dichroic mirror 14, the dichroic mirror 13, and the filter 21 becomes the observation light L2.
  • the observation light L2 that has passed through the filter 21 enters the lens 22 .
  • the lens 22 is an imaging lens and forms an image of the sample S on the light receiving surface of the camera 23 .
  • the camera 23 is a two-dimensional photodetector such as a (Charge Coupled Device) camera or a CMOS (Complementary Metal Oxide Semiconductor) image sensor. Therefore, the camera 23 captures a two-dimensional optical image of the sample S.
  • the processing device 60 is an information processing device such as a personal computer.
  • the camera 23 outputs imaging data of the optical image of the sample S to the processing device 60 .
  • the processing device 60 stores the imaging data of the optical image in a memory or the like. That is, the processing device 60 stores luminance data corresponding to the amount of light received for each pixel of the camera 23 .
  • the processing device 60 has a display or the like that displays an optical image.
  • the user can extract the ROI of the sample S by checking the optical image captured by the camera 23 . For example, the user can select the ROI on the display screen of processing device 60 . This identifies the coordinates in the optical image.
  • the dichroic mirror 14 may be arranged so that it can be inserted into and removed from the optical path. When observing the optical image of the sample S, the dichroic mirror 14 may be removed from the optical path. In other words, the dichroic mirror 14 should be inserted into the optical path during spectroscopic measurement.
  • the spectroscopic measurement optical system 30 is an optical system from the sample S to the photodetector 55 of the spectroscope 50 . That is, the spectroscopic measurement optical system 30 guides the signal light L3 generated by the sample S to the photodetector 55 .
  • the spectroscopic measurement optical system 30 has a lens 15 , a dichroic mirror 14 , a filter 31 , a lens 32 , a fiber unit 40 and a spectroscope 50 .
  • the signal light L3 generated by the sample S is incident on the lens 15.
  • Signal light L3 refracted by lens 15 enters dichroic mirror 14 .
  • the dichroic mirror 14 transmits light of laser wavelength.
  • the dichroic mirror 14 reflects the signal light L3 having a wavelength different from that of the illumination light L1 toward the filter 31 .
  • the signal light L 3 reflected by the dichroic mirror 14 enters the filter 31 .
  • the filter 31 is an optical filter and transmits part of the light from the sample S.
  • the filter 31 is a wavelength filter that transmits or blocks light depending on the wavelength.
  • the filter 31 blocks light of the laser wavelength of the light source 11 and transmits light of a predetermined wavelength band.
  • the spectrometer 50 which will be described later, can spectroscopically measure the signal light L3 having a wavelength different from the laser wavelength.
  • the signal light L3 that has passed through the filter 31 is incident on the lens 32 .
  • the lens 32 is an imaging lens and forms an image of the sample S on the incident end of the fiber unit 40 . That is, the incident end surface of the fiber unit 40 is arranged at a position conjugate with the sample S.
  • the fiber unit 40 has a plurality of fibers and guides the incident signal light L3 to the spectroscope 50 .
  • the fiber unit 40 includes an incident side holder 41 , an exit side holder 42 and a connecting portion 43 .
  • the fiber unit 40 is a bundle fiber in which a plurality of fibers are bundled.
  • a fiber unit 40 is arranged in the optical path from the sample S to the spectroscope 50 .
  • the fiber unit 40 includes an incident-side holder 41, an exit-side holder 42, and a connection portion 43.
  • the fiber arrangement differs between the incident end face and the exit end face of the fiber unit 40.
  • the incident-side holder 41 is, for example, a cylindrical holder that accommodates a plurality of fibers inside.
  • the incident side holder 41 fixes a plurality of fibers on the incident end face side of the fiber unit 40 . Therefore, on the incident end side of the fiber unit 40, a plurality of fibers are arranged adjacent to each other. Specifically, a plurality of fibers are arranged in a close-packed arrangement.
  • the output side holder 42 is, for example, a cylindrical holder, and accommodates a plurality of fibers inside.
  • the output side holder 42 fixes a plurality of fibers on the output end face side of the fiber unit 40 .
  • On the output end side of the fiber unit 40 a plurality of fibers are arranged in a multi-line pattern with an interval. Therefore, spectrometry can be performed with a multifocal optical system.
  • the connecting portion 43 connects the incident side holder 41 and the emitting side holder 42 .
  • the connecting portion 43 is a flexible portion arranged between the incident side holder 41 and the emitting side holder 42 .
  • the configuration of the fiber unit 40 will be described later.
  • the output end of the fiber unit 40 is arranged just before the spectroscope 50 .
  • the signal light L3 propagated through the fibers of the fiber unit 40 is emitted from the emission end of the fiber unit 40 and enters the spectroscope 50 .
  • a signal light L3 from the sample S is incident on the spectroscope 50 via the fiber unit 40 .
  • the spectroscope 50 includes a lens 51, a grating 52, a lens 54, and a photodetector 55.
  • a signal light L3 from the fiber unit 40 is incident on the lens 51 .
  • the lens 51 refracts the signal light L3 so that it becomes a parallel light beam.
  • Signal light L3 from lens 51 enters grating 52 .
  • the grating 52 is a wavelength dispersion element that disperses the signal light L3 according to wavelength.
  • the grating 52 has a diffraction angle according to wavelength.
  • the spectral direction of the signal light L3 by the grating 52 is assumed to be the X direction.
  • the grating 52 is shown as a transmissive diffraction grating, the signal light L3 reflected by the grating 52 enters the lens 54.
  • the wavelength dispersion element is not limited to a transmission type diffraction grating, and may be a reflection type diffraction grating, a prism, or the like.
  • the signal light L3 wavelength-dispersed by the grating 52 enters the lens 54 .
  • the lens 54 converges the signal light L3 on the light receiving surface of the photodetector 55 .
  • the photodetector 55 is a two-dimensional array photodetector such as a CCD (Charge Coupled Device) camera or a CMOS (Complementary Metal Oxide Semiconductor) image sensor.
  • the photodetector 55 has, for example, a plurality of pixels arranged along the X direction and the Y direction.
  • the X direction and the Y direction are directions perpendicular to the optical axis of the optical system. That is, the XY plane is a plane perpendicular to the optical axis.
  • the X coordinate of the photodetector 55 corresponds to the wavelength and spatial position of the signal light L3.
  • An image of the output end of the fiber unit 40 is formed on the light receiving surface of the photodetector 55 .
  • the photodetector 55 can capture a spectroscopic image of the sample S spectroscopically separated by the spectroscope 50 .
  • FIG. 2 is a diagram schematically showing the configuration of the fiber unit 40. As shown in FIG.
  • the fiber unit 40 has a plurality of fibers 45.
  • the incident side holder 41 fixes the plurality of fibers 45 on the incident end face 411 of the fiber unit 40 .
  • a plurality of fibers 45 are arranged adjacent to each other on the incident end face 411 of the fiber unit 40 .
  • the incident ends of the fibers 45 are circular, and a plurality of fibers 45 are arranged so that each circle touches the adjacent circle.
  • the incident end surface 411 is positioned conjugate with the sample S. An image of the sample S is formed on the incident end surface 411 .
  • the signal light L3 from one light irradiation spot on the sample enters one or more fibers 45 . By arranging the fibers 45 closer together, the gap between the fibers 45 can be reduced.
  • the area of the sample S that cannot be spectroscopically measured can be reduced. If the signal light L3 from the sample S enters the gap between the fibers 45, the signal light L3 does not reach the spectroscope 50. FIG. By closely arranging the fibers 45, the area of the gaps between the fibers 45 can be reduced. As a result, the area where spectral measurement cannot be performed can be reduced.
  • Non-Patent Document 4 https://www.tem-inc.co.jp/products/detail-29.php discloses a technique for numbering and mapping multiple fibers. Using this technique, the spacing between fibers can be eliminated or reduced. By fusion-bonding the fibers, a honeycomb structure is formed, eliminating gaps between the fibers. Adjacent fibers may also be glued together with an adhesive such as epoxy resin.
  • 1800 fibers 45 are two-dimensionally densely arranged on the incident end surface 411 .
  • a plurality of fibers 45 are arranged in a hexagonal close-packed array. Therefore, one fiber 45 is arranged so as to be in contact with six fibers 45 around it. In other words, each of the plurality of fibers 45 contacts six fibers 45 .
  • the fibers 45 are not arranged in a matrix. That is, the two arrangement directions on the incident end surface 411 are not orthogonal.
  • the incident side holder 41 holds the fibers 45 so that the plurality of fibers 45 are closely arranged on the incident end surface 411 of the fiber unit 40 . That is, the incident side holder 41 fixes the plurality of fibers 45 so that the fibers 45 are arranged adjacent to each other. By doing so, the spectroscopic measurement of the sample S can be performed more appropriately.
  • An image of the output end surface 421 is formed on the light receiving surface of the photodetector 55 .
  • a plurality of fibers 45 are arranged in a multi-line pattern with a gap.
  • the output side holder 42 holds the plurality of fibers 45 so that the plurality of fibers 45 are arranged at intervals.
  • a plurality of fibers 45 are arranged along the X direction and the Y direction on the output end face 421 .
  • a plurality of fibers 45 are arranged in a two-dimensional matrix. The two array directions on the output end face 421 are not orthogonal.
  • a plurality of fibers 45 are arranged at regular intervals in the X direction.
  • a plurality of fibers 45 are arranged at regular intervals in the Y direction. In this way, the output side holder 42 holds the plurality of fibers 45 so that the plurality of fibers 45 are arranged at regular intervals.
  • 1800 fibers 45 are arranged in a 15 ⁇ 120 matrix. That is, the fibers 45 are arranged in a multi-line form of 15 lines, and 120 fibers 45 are included in one line. 120 fibers 45 included in one line are arranged in parallel in the Y direction.
  • 1800 fibers 45 are arranged in a circular area with a diameter of about 2 to 3 cm.
  • a single fiber 45 has a circular diameter of about 45 ⁇ m.
  • a circular area with a diameter of about 90 ⁇ m is detected by the photodetector 55 . That is, a circular area with a diameter of about 90 ⁇ m is enlarged and imaged on the incident end surface 411 of the fiber unit 40 . Therefore, the photodetector 55 simultaneously detects the signal light L3 from this circular area.
  • 1800 fibers 45 are arranged in a rectangular area of about 14 to 15 cm in the X direction and about 6 to 7 cm in the Y direction. It should be noted that multi-slits corresponding to multi-lines may be provided on the output end face 421 .
  • the output side holder 42 fixes the plurality of fibers 45 so that the plurality of fibers 45 are arranged at intervals on the output end face 421 .
  • the density of the plurality of fibers 45 on the incident end face 411 is higher than the density of the plurality of fibers 45 on the exit end face 421 .
  • the fibers 45 are sparsely arranged on the output end face 421 and densely arranged on the incident end face 411 .
  • the density is 60% or more in the region where the fibers 45 are arranged. in short.
  • the fibers 45 are densely arranged so that the fibers 45 occupy 60% or more of the area in which the fibers 45 are arranged.
  • the interstitial area between fibers 45 is preferably less than 40%.
  • the fibers 45 are densely arranged so that the fibers 45 occupy 75% or more of the area in which the fibers 45 are arranged.
  • the area of interstices between fibers 45 is less than 25%.
  • the signal lights L3 emitted from the adjacent fibers 45 enter the photodetector 55 so as not to overlap each other. That is, the signal light L3 from different fibers 45 is detected by different pixels of the photodetector 55. FIG. By doing so, the spectral image can be appropriately measured.
  • the signal light L3 emitted from the fiber unit 40 as described above is split by the spectroscope 50 .
  • the spectral direction of the spectroscope 50 is defined as the X direction.
  • a pixel address in the X direction on the photodetector 55 indicates the spectral wavelength and spatial position.
  • a pixel address in the Y direction indicates a position in a multiline.
  • FIG. 3 is a diagram showing an image of the light receiving surface of the photodetector 55.
  • the pixels of the photodetector 55 are arranged in the X direction and the Y direction.
  • FIG. 3 schematically shows fibers 45 included in one line.
  • the arrangement direction of the fibers 45 included in one line is parallel to the Y direction.
  • the arrangement direction of the lines is parallel to the X direction. Therefore, the arrangement direction of the fibers 45 and the arrangement direction of the pixels are parallel.
  • the spectral direction of the grating 52 is the X direction.
  • the spectral data of one line of fiber 45 is measured in a strip-shaped detection area on the light receiving surface.
  • the detection area where the spectral data of the fiber 45 included in the first line is measured is indicated as a detection area D1.
  • the detection regions where the spectral data of the fiber 45 on the 2nd to 15th lines are measured are shown as detection regions D2 to D15, respectively. Since 120 fibers 45 are included in one line, 120 spectral data can be measured in one detection area D1.
  • the detection areas D1 to D15 are arranged so as not to overlap each other.
  • the detection area D1 and the detection area D2 are shifted in the Y direction.
  • the spectroscope 50 disperses the signal light on the long wavelength side to the +X side and the signal light L3 on the short wavelength side to the -X side.
  • the pixel address corresponding to the longest wavelength in the detection area D1 is on the -X side of the pixel address corresponding to the shortest wavelength in the detection area D2.
  • the filter 31 limits the wavelength range of the signal light L3 so that the detection areas D1 to D15 do not overlap.
  • the distance in the X direction between the fibers 45 on the output end face 421 is set to a predetermined value or more.
  • the fibers 45 are arranged apart from each other on the output end face 421 .
  • fibers 45 included in one line are spaced apart in the Y direction. Therefore, the signal light L3 from the adjacent fiber 45 is incident on pixels with different Y-direction addresses. Signal light L3 from each fiber 45 is detected by different pixels. In other words, the signal light L3 from the other fiber 45 does not enter the pixel into which the signal light L3 from one fiber 45 has entered.
  • the signal light L3 from each fiber 45 is detected without overlapping. Therefore, the Raman spectrum of a plurality of points of the sample S can be measured with a single shot (one frame) of the photodetector 55 without scanning the illumination light L1. This makes it possible to detect Raman scattered light from a two-dimensional area on a plane perpendicular to the optical axis.
  • part of the signal light from each fiber 45 may be detected so as to overlap.
  • the original spectrum can be recovered by mathematical techniques such as compressed sensing techniques.
  • the signal light L3 from one fiber 45 can be wavelength-dispersed to more pixels. Therefore, the wavelength range that can be spectroscopically measured can be widened. Alternatively, wavelength resolution can be improved.
  • the signal light L3 from each fiber 45 can be spectroscopically measured independently. It becomes possible to spectroscopically measure the signal light L3 from 1800 points of the sample S at the same time. In other words, a two-dimensional spectral image of 1800 pixels can be captured in a short time.
  • a signal light L3 from an arbitrary light irradiation spot (one spot) on the sample S is incident on one fiber 45 .
  • the spectrum of the signal light L3 emitted from one fiber 45 corresponds to the spectrum data of one light irradiation spot (one place) on the sample S.
  • the fiber unit 40 has 1800 fibers 45 . Therefore, the spectrometer 1 can detect 1800 Raman spectra. That is, the spectrometer 1 can spectroscopically measure the signal lights L3 from 1800 points on the sample S, respectively. Then, the processing device 60 generates a spectroscopic image of the sample S based on the detection signal of the photodetector 55 . That is, a spectral image is generated based on 1800 spectral data. The process of generating a spectral image by the processing device 60 will be described below.
  • the incident side holder 41 and the emitting side holder 42 are connected by the connecting portion 43 .
  • a plurality of fibers 45 are bundled in the connecting portion 43 so as to be deformable.
  • the correspondence relationship between the position of each fiber 45 on the incident end face 411 and the position of each fiber 45 on the exit end face 421 is known.
  • the position of the incident end face 411 and the position of the exit end face 421 of one fiber 45a are associated with each other.
  • the position of the incident end face 411 of another fiber 45b and the position of the exit end face 421 are associated with each other.
  • the processing device 60 stores information indicating the layout relationship of each fiber 45 .
  • the processor 60 stores the correspondence relationship between the position of the fiber 45 on the incident end face 411 and the position on the exit end face 421 .
  • the processing device 60 generates a spectral image from the detection result of the photodetector 55 by referring to the arrangement relationship of each fiber 45 .
  • the processor 60 rearranges the detection data for each pixel of the photodetector 55 so as to match the fiber arrangement on the incident end face 411 .
  • the processing device 60 constructs a two-dimensional spectral image by two-dimensional mapping.
  • the fibers 45 are arranged more densely on the incident end face 411 than on the outgoing end face 421 . Therefore, the photodetector 55 can detect the signal light L3 from more points on the sample S. If the position on the sample corresponds to the area where the fibers 45 are closely arranged, the signal light propagates through one of the fibers 45 and is detected by the photodetector 55 .
  • the spectroscope 50 can spectroscopically measure the signal lights L3 from the plurality of fibers 45 at the same time. That is, the signal light L3 emitted from the fiber 45 is detected independently of the signal light L3 emitted from the other fibers 45. FIG. This allows the spectroscope 50 to spectroscopically measure the signal light L3 from a larger number of fibers 45 .
  • the spectroscopic measurement device 1 can capture a two-dimensional spectroscopic image without scanning laser light. Therefore, it becomes possible to measure a Raman spectroscopic image in a short time. Furthermore, since the Raman spectrum can be measured in a short period of time, measurement with low phototoxicity is possible. Since there is no need to label with a fluorescent substance or the like, label-free spectroscopic measurement is possible. Also, the signal light L3 from each fiber 45 is separated and detected. By doing so, spectrometry can be performed with a high SN (Signal to Noise) ratio.
  • an optical image is taken by optical observation of the camera 23 before spectroscopic measurement.
  • observation illumination light L4 from the observation illumination light source 18 illuminates the entire field of view of the objective lens.
  • the user or the processing device 60 extracts a plurality of points on the sample as ROI based on the optical image.
  • processor 60 displays an optical image on a monitor.
  • the user designates an area of interest with a mouse or the like while viewing the optical image on the monitor.
  • Processing device 60 stores the coordinates of the designated area. This extracts the ROI.
  • observation illumination light L4 for capturing an optical image is light from a light source different from the illumination light L1 used during spectroscopic measurement.
  • the observation illumination light source 18 and the light source 11 can be switched between when capturing an optical image and when measuring the spectroscopy.
  • the spatial light modulator 12 controls the beam of the illumination light L1 from the light source 11 so as to selectively illuminate the ROI on the sample S.
  • the spatial light modulator 12 modulates the illumination light L1 so that the illumination light L1 is incident only on the ROI. In other words, the spatial light modulator 12 modulates the illumination light L1 so that the illumination light L1 is not applied to areas other than the ROI. As a result, multiple points extracted as ROIs are illuminated at the same time.
  • a processing device 60 generates a binary image for controlling the spatial light modulator 12 .
  • a binary image for example, it is 1 at positions that are ROIs and 0 at positions that are not ROIs.
  • the processor 60 outputs a control binary image to the spatial light modulator 12, which is an LCOS device.
  • the spatial light modulator 12 has a plurality of control pixels and diffracts incident light. The spatial light modulator 12 can control the phase of diffracted light for each pixel.
  • the spatial light modulator 12 By using the spatial light modulator 12, it is possible to form a multi-focus for arbitrary multiple points on the sample S. That is, since the illumination light L1 is focused only on the ROI, it is possible to prevent Raman scattered light from being generated outside the ROI. This allows measurements with a high SN ratio. In addition, phototoxicity to the sample S can be reduced because only the ROI is irradiated with light.
  • the connective tissue and nerves of the biological sample are extracted as ROIs. Therefore, the spatial light modulator 12 modulates the illumination light L1 so that only the connections and nerves are selectively illuminated.
  • a signal light L3 from the ROI propagates to the spectroscope 50 via the fiber unit.
  • the spectroscope 50 spectroscopically measures the signal light L3 from the location illuminated by the illumination light L1.
  • the processing device 60 generates a spectral image based on the spectrum measurement result of the spectroscope 50 .
  • the processing device 60 decodes the spectral measurement results and segment images. Therefore, the processing device 60 generates images based on the segment images of the optical image in areas other than the ROI, and generates images based on the spectral measurement results in the ROI. Thereby, a more appropriate spectral image can be generated and displayed.
  • Spectroscopic images contain spectral information in ROIs.
  • FIG. 5 is a diagram for explaining the fiber unit 40 when the illumination light L1 selectively illuminates only the ROI.
  • FIG. 5 shows an incident end face 411 and an exit end face 421 of the fiber unit 40 . Further, a light receiving surface image of the photodetector 55 that has detected the signal light L3 emitted from the fiber unit 40 is shown.
  • the fiber 45 corresponding to the ROI is shown as the fiber 451
  • the fiber 45 corresponding to the area other than the ROI is shown as the fiber 452.
  • An image of the ROI is formed on the incident end surface 411 .
  • the position of the fiber 451 on the incident end face 411 corresponds to the ROI.
  • a signal light L3 from the ROI enters the fiber 451 . Areas other than the ROI are not illuminated with the illumination light L1. Therefore, the signal light L3 is not generated in regions other than the ROI. Therefore, the signal light L3 does not enter the fiber 452 .
  • the spectroscope 50 splits the signal light L3.
  • the light receiving surface of the photodetector 55 splits the signal light L3 in the X direction.
  • the signal light L3 from each fiber 451 is detected so as not to overlap on the light receiving surface. Therefore, the signal light L3 from each point of the sample S can be spectroscopically measured appropriately. Since light from regions other than the ROI can be reduced, noise light can be suppressed. Therefore, measurement with a high SN ratio is possible. Therefore, it is possible to generate an appropriate spectroscopic image with short-time measurement.
  • the array direction of the multilines and the array direction of the pixels are parallel, but the array direction of the multilines and the array direction of the pixels may not be parallel. Also, the spectral direction and the pixel array direction may not be parallel.
  • FIG. 6 is a diagram showing images of the incident end face 411 and the exit end face 421.
  • FIG. A plurality of fibers 45 are arranged close to each other on the incident end face 411 .
  • a plurality of fibers 45 are in a close-packed array.
  • a plurality of fibers 45 are arranged in a multi-line arrangement on the output end face 421 .
  • 15 lines M1 to M15 are formed on the output end face 421.
  • the spectroscopic measurement method includes steps of illuminating a sample with illumination light from a light source, and inputting signal light from the sample into a fiber unit from an incident end face in which a plurality of fibers are arranged adjacently.
  • Example 1 will be described with reference to FIG. FIG. 7 shows an optical image of the sample S and the results of spectroscopic measurement of Raman scattered light.
  • the spectral measurement result indicates the amount of light detected by each pixel of the photodetector 55 .
  • the sample S is polystyrene.
  • the spectral illumination optical system 10 selectively illuminates the laser spots A and B as an ROI. Then, the signal light L3 from the laser spots A and B enters the spectroscope 50 via the fiber unit 40.
  • Example 2 will be described with reference to FIG. FIG. 8 shows an optical image of the sample S and the results of spectroscopic measurement of Raman scattered light.
  • sample S contains polystyrene and calcium carbonate (CaCO 3 ).
  • Spot 1 and spot 2 in FIG. 8 are ROIs.
  • polystyrene and calcium carbonate are each extracted as ROI.
  • spot 1 corresponds to polystyrene and spot 2 corresponds to calcium carbonate.
  • the signal light L3 from the spot 1 and the signal light L3 from the spot 2 are incident on different pixels on the light receiving surface. Therefore, each Raman spectrum can be measured at different pixels of the photodetector 55 .
  • the peak wavelength of the Raman spectrum of polystyrene is different from the peak wavelength of the Raman spectrum of calcium carbonate.
  • FIG. 9 is an image showing ROI extraction and ROI illumination with selective illumination of the ROI.
  • the sample S is polystyrene.
  • a ROI is extracted from the optical image of the sample S.
  • the ROI is then selectively illuminated.
  • FIG. 9 shows an image of the incident end face during ROI illumination and the results of spectroscopic measurement. Signal lights from a plurality of points on the sample S are detected by different pixels.
  • the signal light is mainly Raman scattered light, but the signal light may be light other than Raman scattered light. Therefore, the spectroscopic measurement apparatus according to this embodiment may be a spectroscopic measurement apparatus other than Raman spectroscopy.
  • it may be a spectrometer that detects fluorescence excited by excitation light, or a spectrometer that measures an ultraviolet absorption spectrum or a near-infrared absorption spectrum. These spectrometers can also measure spectra with a high SN ratio.
  • it is suitable for a spectroscopic measurement device that requires high-speed measurement and repeated measurement.
  • a biological sample or a medical sample can be used as the sample S to be measured.
  • the sample S can be a sample for pathological diagnosis, a drug discovery sample, foods, cosmetics, advanced materials, devices, and the like. Spectroscopic measurement results can be used to assist in quality control. Furthermore, various samples such as field science (space, ocean, ecology) can be used as the sample S.
  • the endoscope may be a rigid endoscope or a flexible endoscope, and may be a medical endoscope or an industrial endoscope.
  • FIG. 10 is a diagram showing measurement results of a biological sample.
  • an optical image obtained by imaging adipose tissue as a sample and its spectroscopic measurement result are shown.
  • a ROI is extracted from the optical image.
  • FIG. 10 shows the ROI as a laser irradiation spot. Then, the ROI is selectively irradiated with laser light serving as excitation light.
  • Non-transitory computer-readable media include various types of tangible storage media. Examples of non-transitory computer-readable media include magnetic recording media (eg, flexible discs, magnetic tapes, hard disk drives), magneto-optical recording media (eg, magneto-optical discs), CD-ROMs (Read Only Memory), CD-Rs, Includes CD-R/W, semiconductor memory (eg, mask ROM, PROM (Programmable ROM), EPROM (Erasable PROM), flash ROM, RAM (Random Access Memory)).
  • magnetic recording media eg, flexible discs, magnetic tapes, hard disk drives
  • magneto-optical recording media eg, magneto-optical discs
  • CD-ROMs Read Only Memory
  • CD-Rs Includes CD-R/W
  • semiconductor memory eg, mask ROM, PROM (Programmable ROM), EPROM (Erasable PROM), flash ROM, RAM (Random Access Memory)
  • the program may also be delivered to the computer by various types of transitory computer readable media.
  • Examples of transitory computer-readable media include electrical signals, optical signals, and electromagnetic waves.
  • Transitory computer-readable media can deliver the program to the computer via wired channels, such as wires and optical fibers, or wireless channels.

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