WO2020037837A1 - Appareil d'imagerie 3d basé sur une transformation de l'espace k et procédé d'imagerie associé - Google Patents

Appareil d'imagerie 3d basé sur une transformation de l'espace k et procédé d'imagerie associé Download PDF

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Publication number
WO2020037837A1
WO2020037837A1 PCT/CN2018/114488 CN2018114488W WO2020037837A1 WO 2020037837 A1 WO2020037837 A1 WO 2020037837A1 CN 2018114488 W CN2018114488 W CN 2018114488W WO 2020037837 A1 WO2020037837 A1 WO 2020037837A1
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Prior art keywords
sample
dimensional
light
motorized stage
dimensional imaging
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PCT/CN2018/114488
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English (en)
Chinese (zh)
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张雪丹
刘诚
朱健强
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中国科学院上海光学精密机械研究所
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Publication of WO2020037837A1 publication Critical patent/WO2020037837A1/fr

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    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/39—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using tunable lasers
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/4788—Diffraction
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/4788—Diffraction
    • G01N2021/479—Speckle

Definitions

  • the invention relates to the field of three-dimensional imaging, in particular to a three-dimensional imaging device based on K-space transformation and an imaging method thereof.
  • Three-dimensional imaging technology has developed rapidly due to its wide reference. This technology has a great role in the observation of biological samples and tumor diagnosis.
  • There are many methods used in the field of three-dimensional imaging such as structured light illumination technology, confocal scanning, coherence tomography, flake illumination microscopy, and coherence tomography. These technologies are able to reflect the internal structure of the sample, including information such as the reflectance of the sample and the concentration of the luminescent material.
  • Structured light three-dimensional imaging technology uses a carrier frequency stripe to illuminate the object, records the deformed stripes, and then digitally demodulates the reconstructed three-dimensional image from the deformed fringe map obtained to reconstruct the three-dimensional image of the measured object. The shadow effect of the focal part.
  • the most widely used 3D imaging technology is confocal scanning technology.
  • This technology uses pinholes to block the passage of light from the out-of-focus portion, thereby eliminating the effects of shadows from the out-of-focus sample.
  • Optical coherence tomography can use gratings and two-dimensional scanning galvanometers to achieve high lateral and vertical resolution.
  • the flake light illumination microscopy technology uses a flake light source to illuminate the sample, and at the same time collects reflected light in a direction perpendicular to the optical path. Since the illumination light is a flake light, the collected reflected light is the axial distribution of the sample. Since the lighting system and the acquisition system are not co-axial systems, this technology is difficult to implement.
  • the method proposed by this patent has high imaging rate, spatial resolution, simple structure, and easy construction and assembly.
  • the purpose of the present invention is to overcome the above-mentioned shortcomings of the prior art, and provide a three-dimensional imaging method with high imaging speed, high resolution, simple structure, and easy construction and assembly.
  • a three-dimensional imaging device based on K-space transformation which is characterized by comprising a laser, a lens, a first beam splitting prism, a first cylindrical lens, a spot detector, a computer, a second beam splitting prism, a microscope objective lens, and a place for a sample to be measured.
  • the laser light emitted by the laser (1) passes through the lens (2) and becomes a parallel beam.
  • the beam is transmitted and reflected by the first beam splitting prism (3), that is, a detection beam and a reference beam.
  • the detection beam passes through the first
  • a cylindrical lens (4) is focused into a sheet-shaped illumination light, and is condensed into a thinner sheet-shaped sheet-shaped illumination light by a second spectroscopic prism (7) and a microscope objective lens (8) in order, and irradiates the sample to be measured. After the reflection of the sample is measured, the original path is returned, transmitted through the micro objective lens (8), and then incident on the second beam splitting prism (7);
  • the reference beam After the reference beam is reflected by the second reflector (14), a sheet-shaped reference light is formed through the second cylindrical lens (13), and after being reflected by the first reflector (12), it passes through the microscope objective lens group (11) in order. ) And the second spectroscopic prism (7) are transmitted, and together with the detection beam reflected by the second spectroscopic prism (7), they are incident on the spot detector (5), and the computer (6) and the spot detector are respectively (5) Connected to the one-dimensional motorized stage (9).
  • the computer (6) controls the movement of the one-dimensional motorized stage (9), so that the sample to be measured is moved in a direction perpendicular to the optical axis according to a preset moving step l, and each time the movement is performed, the light spot detector collects and repeats the steps 3 , 4, 5 until moving n times, collect 3n scattered light spots;
  • the intensity of the light spot recorded by the radon spot detector is input into the computer, and the computer performs three-dimensional imaging of the sample to be measured.
  • the spot intensity recorded by the spot detector is input to a computer, and the computer uses the spot data to perform three-dimensional imaging of the sample to be measured.
  • a computer is used to calculate the 3n scattered light version recorded by the spot detector.
  • the calculation process is as follows:
  • Step 7.2 Read in the three scattered light spots recorded by the n-th movement of the one-dimensional motorized stage;
  • Step 7.3 Use the sum of the 3rd scattered light spot pattern of the nth time and the sum of the 1st and 2nd scattered light spot patterns of the nth time to make a difference;
  • Step 7.4 Perform a discrete Fourier transform on the image obtained in step 7.3, and then intercept the first-level spectrum and translate it to the center of the image;
  • Step 7.5 Perform inverse discrete Fourier transform on the image obtained in step 7.4, and then use the Fresnel propagation formula to propagate to the center plane of the sample, and the propagation distance is: D + L 1 + L 2 ;
  • Step 7.6 Perform a discrete Fourier transform on the image obtained in step 7.5;
  • Step 7.7 use the formula Obtain the corresponding position of the wavefront on the optical axis, where p is the axial position, m is the lateral position, n is the longitudinal position, ⁇ is the laser wavelength, ⁇ k x is the lateral spatial frequency, ⁇ k y is the longitudinal spatial frequency, and ⁇ k z Axial space frequency
  • Step 7.8 Move the corresponding amplitude and phase of the data (m, n) to (m, p), thereby projecting the wavefront to a plane parallel to the optical axis;
  • Step 7.9 Perform a discrete inverse Fourier transform on the result obtained in step 7.8;
  • This method does not require scanning in order to achieve axial imaging, and a depth map of the sample to be measured can be obtained by one acquisition;
  • This method can eliminate mutual crosstalk between different levels and shield the projection of the blurred position.
  • FIG. 1 is a schematic diagram of an apparatus for a three-dimensional imaging method based on K-space transformation according to the present invention.
  • FIG. 1 is a schematic diagram of a three-dimensional imaging method based on K-space transformation.
  • a laser with a wavelength of 632.8 nm emitted by laser 1 passes through lens 2 to become a parallel light with a flat phase, and then passes through the light.
  • the beam splitting prism 3 with a ratio of 1: 1 splits the beam to produce two parallel beams of equal intensity, which are a detection beam and a reference beam, respectively.
  • the detection beam is focused by the cylindrical lens 4 into a sheet-shaped illumination light and passes through a microscope objective lens. 5.
  • the sample holder to be tested is controlled by a one-dimensional motorized stage 9, and the light in the reference optical path is split by a beam splitter and passed through After being reflected by the reflecting mirror 14, the sheet-shaped reference light is formed through the cylindrical lens 13, and after being reflected by the reflecting mirror 12, the reflected light irradiated on the sample holder 10 to be measured by the micro objective lens group 11 and the detection light passes through the beam splitting prism. 7. Record by the spot detector 5 and transfer the recorded data to the computer 6.
  • the laser (1) emits a laser with a wavelength of 632.8 nm and passes through the lens (2) to become a parallel beam.
  • the beam is split into a transmitted beam and a reflected beam by a first beam splitting prism (3) with a beam splitting ratio of 1: 1, that is, detection.
  • Light beam and reference light beam, the detection beam is focused by the first cylindrical lens (4) into sheet-like illumination light, and then passes through a second beam splitting prism (7) and a microscope objective lens (8) in order of 1: 1.
  • the thinner sheet-shaped illumination light condensed into a thin sheet is irradiated on the sample to be measured, and after being reflected by the sample to be measured, it returns to the original path, passes through the microscope objective lens (8), and enters the second beam splitting prism (7);
  • the reference beam After the reference beam is reflected by the second reflector (14), a sheet-shaped reference light is formed through the second cylindrical lens (13), and after being reflected by the first reflector (12), it passes through the microscope objective lens group (11) in order. ) And the second spectroscopic prism (7) are transmitted, and together with the detection beam reflected by the second spectroscopic prism (7), they are incident on the spot detector (5), and the computer (6) and the spot detector are respectively (5) Connected to the one-dimensional motorized stage (9).
  • the focal length of the first cylindrical lens 4 and the second cylindrical lens 13 is 40 mm
  • the linear distance between the cylindrical lens 4 and the microscope objective lens 8 is 48.83 mm
  • the microscope objective lens 8 is a 10x objective lens
  • the microscope objective lens 8 is clamped to the sample to be measured.
  • the linear distance of the detector 10 is 0.97mm
  • the resolution of the light spot detector 5 is 2048 pixels ⁇ 2048 pixels
  • the minimum unit is 5.5 ⁇ m.
  • the computer (6) controls the movement of the one-dimensional motorized stage (9), so that the sample to be measured is moved in a direction perpendicular to the optical axis according to a preset moving step l, and each time the movement is performed, the light spot detector collects and repeats the steps 3 , 4, 5 until moving n times, collect 3n scattered light spots;
  • the intensity of the light spot recorded by the radon spot detector is input into the computer, and the computer performs three-dimensional imaging of the sample to be measured.
  • the spot intensity recorded by the spot detector is input to a computer, and the computer uses the spot data to perform three-dimensional imaging of the sample to be measured.
  • a computer is used to calculate the 3n scattered light version recorded by the spot detector.
  • the calculation process is as follows:
  • Step 7.2 Read in the three scattered light spots recorded by the n-th movement of the one-dimensional motorized stage;
  • Step 7.3 Use the sum of the 3rd scattered light spot pattern of the nth time and the sum of the 1st and 2nd scattered light spot patterns of the nth time to make a difference;
  • Step 7.4 Perform a discrete Fourier transform on the image obtained in step 7.3, and then intercept the first-level spectrum and translate it to the center of the image;
  • Step 7.5 Perform inverse discrete Fourier transform on the image obtained in step 7.4, and then use the Fresnel propagation formula to propagate to the center plane of the sample, and the propagation distance is: D + L 1 + L 2 ;
  • Step 7.6 Perform a discrete Fourier transform on the image obtained in step 7.5;
  • Step 7.7 use the formula Obtain the corresponding position of the wavefront on the optical axis, where p is the axial position, m is the lateral position, n is the longitudinal position, ⁇ is the laser wavelength, ⁇ k x is the lateral spatial frequency, ⁇ k y is the longitudinal spatial frequency, and ⁇ k z Axial space frequency
  • Step 7.8 Move the corresponding amplitude and phase of the data (m, n) to (m, p), thereby projecting the wavefront to a plane parallel to the optical axis;
  • Step 7.9 Perform a discrete inverse Fourier transform on the result obtained in step 7.8;
  • the experimental results show that the device of the present invention successfully realizes three-dimensional imaging of the sample.
  • the device uses scanning to record 3n scattered light spots, and the computer calculates and displays the three-dimensional structural characteristics of the sample. This method is not limited to the size of the light spot detector. Less affected by the environment, the device has a simple structure, high measurement resolution, and fast imaging speed, which meets the requirements for three-dimensional inspection of optical components.

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  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pathology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Optics & Photonics (AREA)
  • Length Measuring Devices By Optical Means (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

La présente invention concerne un procédé d'imagerie 3D basé sur une transformation de l'espace K, le procédé comprenant l'utilisation d'une lumière en forme de feuille pour éclairer un échantillon le long de la direction d'un axe optique, et l'utilisation d'une source de lumière en forme de feuille identique pour interférer avec la lumière d'éclairage ; l'enregistrement d'un interférogramme et l'acquisition à partir de celui-ci d'informations d'amplitude complexes (comprenant l'amplitude et la phase) de l'échantillon éclairé dans le plan focal ; des informations de spectre de fréquence spatiale de l'échantillon peuvent être acquises au moyen d'une transformation de Fourier discrète ; l'utilisation d'un procédé de projection pour calculer des informations de spectre de fréquence spatiale perpendiculaires au plan de l'échantillon ; l'acquisition, au moyen d'une transformation inverse de Fourier discrète, d'informations de force perpendiculaires au plan de l'échantillon ; et l'acquisition, au moyen d'un balayage, d'informations de structure 3D finales de l'échantillon. Le procédé présente un taux d'acquisition élevé et une haute résolution, en particulier pour l'acquisition d'informations axiales, et doit effectuer une acquisition une seule fois pour mettre en oeuvre une imagerie.
PCT/CN2018/114488 2018-08-21 2018-11-08 Appareil d'imagerie 3d basé sur une transformation de l'espace k et procédé d'imagerie associé WO2020037837A1 (fr)

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CN201810954592.9A CN109085137B (zh) 2018-08-21 2018-08-21 基于k空间变换的三维成像装置及其成像方法
CN201810954592.9 2018-08-21

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