WO2023130636A1 - Module, dispositif et procédé de microscopie à contraste de phase basee sur un cristal uniaxe - Google Patents

Module, dispositif et procédé de microscopie à contraste de phase basee sur un cristal uniaxe Download PDF

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Publication number
WO2023130636A1
WO2023130636A1 PCT/CN2022/091660 CN2022091660W WO2023130636A1 WO 2023130636 A1 WO2023130636 A1 WO 2023130636A1 CN 2022091660 W CN2022091660 W CN 2022091660W WO 2023130636 A1 WO2023130636 A1 WO 2023130636A1
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WIPO (PCT)
Prior art keywords
polarized light
circularly polarized
handed circularly
polarizer
phase contrast
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PCT/CN2022/091660
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English (en)
Chinese (zh)
Inventor
朱文国
赵梦婷
余健辉
陈哲
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暨南大学
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Publication of WO2023130636A1 publication Critical patent/WO2023130636A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/14—Condensers affording illumination for phase-contrast observation
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/0092—Polarisation microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes

Definitions

  • the invention relates to the fields of optical imaging and optical information processing, in particular to a phase contrast microscope module, method and equipment based on uniaxial crystals.
  • optical phase contrast microscopy can not only realize the edge enhancement of object intensity information, but also convert the phase information of transparent objects into intensity patterns. Therefore, optical phase contrast microscopy has broad application prospects in the fields of optical high-contrast imaging, biomedicine, face recognition, and optical simulation computing.
  • optical phase contrast microscopy there are many methods of optical phase contrast microscopy, and the commonly used methods include Zernike phase contrast microscopy, Nomarski differential interference phase contrast imaging, and optical space differential microscopy.
  • Traditional Zernike phase-contrast microscopic imaging and Nomarski differential interference phase-contrast imaging rely on complex modulation in space or spatial frequency domains, resulting in complex systems and difficulties in optical alignment and adjustment.
  • Optical space differential microscopy imaging can realize phase-contrast microscopy by constructing a suitable transfer function and performing differential processing on the light field.
  • Optical spatial differential microscopy has attracted extensive attention due to its wide operating frequency band, isotropic edge enhancement and relatively compact optical system.
  • optical spatial differentiators are mainly based on micro-nano structures such as metal surface plasmons and artificial metasurfaces, which are expensive to manufacture and difficult to produce on a large scale.
  • the object of the present invention is to provide a phase contrast microscope module, equipment and method based on uniaxial crystals to solve the above problems.
  • An embodiment of the present invention provides a phase contrast microscope module based on a uniaxial crystal, which includes:
  • the polarization states of the first polarizer and the second polarizer are orthogonal or parallel, and the uniaxial crystal is arranged between the first polarizer and the second polarizer;
  • the first polarizer is used to convert the received incident light carrying the information of the object to be measured into linearly polarized light polarized along a specified direction;
  • the uniaxial crystal is used to displace the left-handed circularly polarized light and the right-handed circularly polarized light generated by the linearly polarized light in opposite directions based on the photon spin Hall effect and/or angle dispersion effect;
  • the second polarizer is used to perform a sum or difference operation on the left-handed circularly polarized light field and the right-handed circularly polarized light field, so that when the displacement of the left-handed circularly polarized light field and the right-handed circularly polarized light is much smaller than the spot size , the transmitted light field is the first differential of the original incident light field.
  • the uniaxial crystals are yttrium vanadate, lithium niobate, quartz, calcite, BBO uniaxial crystals.
  • the left-handed circularly polarized light and the right-handed circularly polarized light with opposite spin directions generated by the uniaxial crystal have a lateral displacement caused by photon spin splitting.
  • the optical axis of the uniaxial crystal has a certain inclination angle relative to the beam propagation direction, and the displacement can be adjusted by changing the inclination angle.
  • the first-order differential of the incident light field in one dimension is realized
  • An embodiment of the present invention also provides a uniaxial crystal-based phase-contrast microscope, which includes an illumination module, an imaging module, and the above-mentioned phase-contrast microscope module.
  • the imaging module includes a first objective lens, a first focusing lens and a camera, and the phase contrast microscope module is arranged between the first focusing lens and the camera; the first objective lens is arranged on the first Before focusing the lens.
  • the lighting module includes a light source, a second objective lens, and a second focusing lens; wherein, along the propagation direction of incident light, the second objective lens is arranged between the light source and the second focusing lens; the light source For LED, halogen lamp or laser light source.
  • the phase contrast microscopy equipment further includes an object stage; the object stage is arranged between the first focusing lens and the first objective lens.
  • the embodiment of the present invention also provides a phase contrast microscopy method based on the above uniaxial crystal phase contrast microscopy equipment, which includes:
  • the light source emits incident light, and after the incident light is collimated by the second objective lens and focused by the second focusing lens on the object to be imaged on the stage, the first signal light is formed;
  • the first objective lens collimates the first signal light, it is focused onto the camera by the first focusing lens. Before the signal enters the camera, it passes through the phase contrast microscope module;
  • the first polarizer transforms the received signal light into linearly polarized light polarized along a specified direction
  • the uniaxial crystal is based on the photon spin Hall effect and/or angle dispersion effect, so that the left-handed circularly polarized light and the right-handed circularly polarized light generated by the linearly polarized light are displaced in opposite directions; wherein, the left-handed circularly polarized light and the right-handed circularly polarized light Circularly polarized light has an overlapping region, and the overlapping region contains both left-handed circularly polarized light and right-handed circularly polarized light;
  • the second polarizer performs a sum or difference operation on the left-handed circularly polarized light field and the right-handed circularly polarized light field to obtain the second signal light.
  • the second signal light is the first order differential of the original incident light field;
  • a camera records the second signal light to obtain phase contrast information of the object to be imaged.
  • the embodiment of the present invention utilizes the method of spin optics to realize the spatial differential calculation of the input image, which can be used for edge enhancement of object intensity information and visualization of object phase information.
  • the phase contrast microscopy technology realized by the embodiment of the present invention is more intuitive, more convenient, and more time-saving, and the embodiment of the present invention can be directly embedded in the existing optical microscopy system,
  • the overall implementation cost is low and easy to integrate.
  • Fig. 1 is a schematic structural diagram of a phase contrast microscope module based on a uniaxial crystal provided in the first embodiment of the present invention.
  • Figure 2(a) shows the spin-splitting shift of right-handed polarized light as a function of incident angle.
  • FIG. 3 is a schematic structural diagram of a phase contrast display device based on a uniaxial crystal provided in a second embodiment of the present invention.
  • Figure 4(a)- Figure 4(d) are phase contrast microscopy detection images based on the theoretical calculation of the photon spin Hall effect.
  • Fig. 5(a)-Fig. 5(h) are experimental comparison diagrams of phase-contrast microscopic examination of the resolution plate.
  • Fig. 6(a)-Fig. 4(d) are comparison diagrams of phase-contrast microscopy experiments on onion epidermal cells.
  • Fig. 7 is a schematic flow chart of the phase contrast microscopy method provided by the third embodiment of the present invention.
  • the first embodiment of the present invention provides a phase contrast microscope module 10 based on uniaxial crystal, which includes:
  • the first polarizer 11 is used to transform the received incident light carrying the information of the object to be measured into linearly polarized light polarized along a specified direction.
  • the incident light may be light emitted by an incoherent light source or a coherent light source. Before the incident light reaches the first polarizer 11, it needs to pass through the object to be imaged.
  • the imaged object is a transparent object, so that the incident light carries the information of the object.
  • the uniaxial crystal 13 is used for, based on the photon spin Hall effect and/or angle dispersion effect, causing the left-handed circularly polarized light and the right-handed circularly polarized light generated by the linearly polarized light to be displaced in opposite directions.
  • the photon spin Hall effect when the beam passes through the surface of a non-uniform medium for reflection and refraction, the photons with opposite spin angular momentum will separate from each other in the direction perpendicular to the incident surface, resulting in spin splitting of the beam Phenomenon.
  • the incident wave function can be expressed as: (where s represents the spin state of the particle)
  • the output wave function can be expressed as: In the formula, ⁇ represents the displacement caused by spin splitting.
  • the above formula can be simplified as It can be seen from the above formula that the spatial differential is generated due to the opposite displacement ⁇ of the spin state, that is, the spatial differential calculation is essentially a photon spin Hall effect. Therefore, the optical spatial differential calculation of the input image can be realized by using the photonic spin Hall effect.
  • the uniaxial crystal 13 can be uniaxial crystals such as yttrium vanadate, lithium niobate, quartz, calcite, BBO, etc., of course, it can also be other uniaxial crystals with the same or similar characteristics. Be specific. Taking the uniaxial yttrium vanadate crystal as an example, when a beam of linearly polarized light is incident on the uniaxial yttrium vanadate crystal, a photon spin Hall effect will be generated, in particular, a left-handed circular polarization with opposite spin direction will be generated. Light and right-handed circularly polarized light, the two beams of light with opposite spins produce a spin-split displacement ⁇ in the transverse direction. When the displacement ⁇ is small enough, there will be an overlapping region containing both left-handed circularly polarized light and right-handed circularly polarized light.
  • the displacement ⁇ can be adjusted by changing the inclination angle of the uniaxial crystal 13 relative to the normal of the optical axis, where the direction of the optical axis is consistent with the propagation direction of the incident light.
  • Figure 2(a) shows the spin-splitting displacement of right-handed polarized light as a function of incident angle.
  • Figure 2(b) shows the spin-splitting shift of left-handed polarized light as a function of incident angle.
  • the second polarizer 12 is used to perform a sum or difference operation on the left-handed circularly polarized light field and the right-handed circularly polarized light field, so that when the displacement of the left-handed circularly polarized light field and the right-handed circularly polarized light is much smaller than the spot size
  • the transmitted light field is the first order differential of the original incident light field.
  • the first-order differential in one dimension of the incident light field is realized
  • phase contrast microscope module 10 when the incident light beam is obliquely incident on the surface of the uniaxial crystal 13 at a certain angle, due to the effect of the refractive index gradient, the photon spin Hall effect will be generated to realize the The spatial differentiation of the input image enables edge enhancement of object intensity information and visualization of object phase information.
  • the phase-contrast microscopy technique implemented in this embodiment is more intuitive, more convenient, and more time-saving.
  • the phase-contrast microscope module 10 provided in this embodiment can be directly embedded in existing microscope equipment, and the overall implementation cost is low and easy to integrate. Among them, when applied to microscopic equipment, when the phase contrast microscopic module 10 is not inserted, the microscopic equipment can present a clear image, and when the phase contrast microscopic module is inserted, the microscopic equipment can realize edge enhancement and object phase information visualization.
  • the second embodiment of the present invention provides a phase-contrast microscope device based on uniaxial crystal, which includes an illumination module, an imaging module, and a phase-contrast microscope module 10 according to any of the above-mentioned embodiments.
  • the imaging module includes a first objective lens 20, a first focusing lens 31 and a camera 32, and the phase contrast microscope module 10 is arranged between the first focusing lens 31 and the camera 32; the first objective lens 20 It is arranged in front of the first focusing lens 31.
  • the camera may be a CCD, a CMOS camera, etc., which are not specifically limited in the present invention.
  • the illumination module includes a light source 40, a second objective lens 50, and a second focusing lens 60, and along the direction of incident light propagation, the second objective lens 50 is arranged between the light source 40 and the second focusing lens 60 .
  • the light source 40 is a light source such as LED, halogen lamp or laser.
  • the light source 40 is an incoherent light source, such as an LED light source. Compared with laser lighting, LED lighting is more uniform, and the cost of LED is lower.
  • the phase contrast microscopy equipment further includes an object stage 70, which is arranged between the second focusing lens 60 and the first objective lens 20, and is used to carry the object to be imaged. object.
  • the working principle of the phase contrast microscope is that when the light passes through the object on the stage 70, the images with different details will have a phase difference, and the optical path of each part after focusing with the second focusing lens 60 Different, the light beams are deflected to different degrees.
  • the two groups of light rays are converged by the first focusing lens 31 and recombined on the same optical path, the direct light and the diffracted light will produce light interference during the propagation process, and the phase difference will become Poor amplitude.
  • phase contrast microscopy equipment is used as an imaging module here.
  • phase contrast imaging is the most effective method for imaging transparent samples. It can obtain the outline details of samples that cannot be seen by ordinary intensity imaging.
  • FIG. 4 is an edge detection diagram realized by theoretically calculating the photon spin Hall effect provided by an embodiment of the present invention.
  • Fig. 4 (a) represents the graph on the right-handed component
  • Fig. 4 (b) represents the graph on the left-handed component
  • Fig. 4 (c) represents the graph on the x component
  • Fig. 4 (b) represents the graph on the y component.
  • FIG. 5 is a comparison diagram of edge detection obtained when the stage 70 is a resolution plate.
  • Figure 5(a), (c), (e), (g) are numbers or graphics on the resolution board
  • Figure 5(b), (d), (f), (h) are the corresponding edges Detection map. It can be seen from the comparison that the edges of each number and figure can be clearly seen on the finally obtained edge detection map.
  • Figure 6(a) and Figure 6(c) are unstained onion epidermal cells observed.
  • Figure 6(b) and Figure 6(d) are the edge detection diagrams obtained by performing edge detection on onion epidermal cells, where Figure 6(a) corresponds to Figure 6(b), and Figure 6(c) corresponds to Figure 6(d) , the edge of the onion epidermal cells can be clearly seen by contrast.
  • the third embodiment of the present invention also provides a phase contrast microscopy method based on the phase contrast microscopy equipment of the uniaxial crystal as described above, which includes:
  • the light source emits incident light, and the incident light is collimated by the second objective lens and focused by the second focusing lens onto the object to be imaged on the stage to form the first signal light;
  • the first objective lens collimates the first signal light, it is focused onto the camera by the first focusing lens. Before the signal enters the camera, it passes through the phase contrast microscope module;
  • the first polarizer transforms the received signal light into linearly polarized light polarized along a specified direction
  • the uniaxial crystal causes the left-handed circularly polarized light and the right-handed circularly polarized light generated by the linearly polarized light to be displaced in opposite directions;
  • the second polarizer is used for the sum or difference calculation of the left-handed circularly polarized light field and the right-handed circularly polarized light field, the transmitted light field of the second polarizer is the first-order differential of the original incident light field, and the second signal light;
  • the camera records the second signal light to obtain phase contrast information of the object to be imaged.
  • the left-handed circularly polarized light and the right-handed circularly polarized light with opposite spin directions generated by the uniaxial crystal have a displacement generated by photon spin splitting in the transverse direction, and when the displacement is less than a preset threshold, the left-handed circularly polarized light There is an overlapping region between circularly polarized light and right-handed circularly polarized light.
  • the uniaxial crystal is tilted at a certain angle relative to the normal of the optical axis, and the direction of the optical axis is consistent with the propagation direction of the incident light, and the displacement is adjusted by changing the tilt angle of the uniaxial crystal relative to the normal of the optical axis .
  • the embodiment of the present invention utilizes the method of spin optics to realize the spatial differential calculation of the input image, which can be used for edge enhancement of object intensity information and visualization of object phase information.
  • the phase contrast microscopy technique implemented in the embodiment of the present invention is more intuitive, more convenient, and more time-saving.

Abstract

L'invention concerne un module, un dispositif et un procédé de microscopie à contraste de phase basée sur un cristal uniaxe. Le module comprend : une première plaque de polarisation, une seconde plaque de polarisation et un cristal uniaxe disposé entre celles-ci. Les états de polarisation de la première plaque de polarisation et de la seconde plaque de polarisation sont orthogonaux ou parallèles. La première plaque de polarisation est utilisée pour convertir des informations de transport de lumière incidente d'un objet à détecter en lumière polarisée linéairement ; le cristal uniaxe est utilisé pour produire, sur la base d'un effet Hall de spin photonique et/ou d'un effet de dispersion angulaire, une lumière à polarisation circulaire à gauche et une lumière à polarisation circulaire à droite générées par la lumière à polarisation linéaire pour générer des déplacements dans des directions opposées ; et la seconde plaque de polarisation est utilisée pour effectuer une opération de somme ou de différence sur un champ de lumière à polarisation circulaire à gauche et un champ de lumière à polarisation circulaire à droite, de telle sorte que lorsque les déplacements de la lumière à polarisation circulaire à gauche et de la lumière à polarisation circulaire à droite sont bien inférieurs à une taille de point, un champ de lumière transmise de celle-ci est un différentiel de premier ordre du champ de lumière incidente d'origine. Le présent procédé peut être utilisé pour l'amélioration de bord d'informations de force d'objet et la visualisation d'informations de phase d'objet. Par comparaison avec une technologie de microscopie à contraste de phase classique, la présente invention est plus visuelle, pratique et économe en temps.
PCT/CN2022/091660 2022-01-10 2022-05-23 Module, dispositif et procédé de microscopie à contraste de phase basee sur un cristal uniaxe WO2023130636A1 (fr)

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CN202210021784.0 2022-01-10
CN202210021784.0A CN114527559B (zh) 2022-01-10 2022-01-10 基于单轴晶体的相衬显微模块、设备及方法

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CN116224572B (zh) * 2023-05-09 2023-08-22 中国人民解放军63921部队 偏振片透偏方向对准系统及方法

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CN108629787A (zh) * 2018-04-02 2018-10-09 浙江大学 一种基于光学自旋霍尔效应空间光场微分器的图像边缘提取方法及系统

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CN111505817B (zh) * 2020-04-30 2022-05-20 河北大学 基于偏振编码的相衬显微系统及其成像方法
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Publication number Priority date Publication date Assignee Title
US20040017609A1 (en) * 2002-04-30 2004-01-29 Rainer Danz Arrangement and method for polarization-optical interference contrast
US20050152030A1 (en) * 2003-12-17 2005-07-14 Marine Biological Laboratory Orientation independent differential interference contrast microscopy technique and device
JP2013041142A (ja) * 2011-08-17 2013-02-28 Yokogawa Electric Corp 顕微鏡装置
CN103529542A (zh) * 2013-10-24 2014-01-22 广州粤显光学仪器有限责任公司 偏振光调制相衬显微镜
CN108629787A (zh) * 2018-04-02 2018-10-09 浙江大学 一种基于光学自旋霍尔效应空间光场微分器的图像边缘提取方法及系统

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