WO2024175010A1 - 成像系统、制备方法及取像装置 - Google Patents

成像系统、制备方法及取像装置 Download PDF

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Publication number
WO2024175010A1
WO2024175010A1 PCT/CN2024/077834 CN2024077834W WO2024175010A1 WO 2024175010 A1 WO2024175010 A1 WO 2024175010A1 CN 2024077834 W CN2024077834 W CN 2024077834W WO 2024175010 A1 WO2024175010 A1 WO 2024175010A1
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Prior art keywords
imaging
modulation unit
phase modulation
plane
lens group
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PCT/CN2024/077834
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English (en)
French (fr)
Inventor
洪煦昊
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Nanjing University
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Nanjing University
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Priority to JP2025548002A priority Critical patent/JP2026505530A/ja
Publication of WO2024175010A1 publication Critical patent/WO2024175010A1/zh
Priority to US19/300,715 priority patent/US20250377526A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B13/00—Optical objectives specially designed for the purposes specified below
    • G02B13/0095—Relay lenses or rod lenses
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G02B21/0056—Optical details of the image generation based on optical coherence, e.g. phase-contrast arrangements, interference arrangements
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/02—Objectives
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • 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
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/362—Mechanical details, e.g. mountings for the camera or image sensor, housings
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements

Definitions

  • the present invention relates to the field of optical imaging technology, and in particular to an imaging system, a preparation method and an imaging device.
  • phase factor of the sample plays a key role.
  • mainstream methods include traditional phase contrast microscopes, Hoffman phase contrast microscopes, differential interference microscopes, etc. to highlight the phase information of the sample to facilitate the observation of weak contrast samples.
  • the imaging resolution of the spiral phase contrast imaging system based on the 4f system is not high, and there is a certain optical distortion.
  • the original imaging optical path needs to be adjusted, and it is not easy to be directly installed in a commercial optical imaging system.
  • an improved imaging system, preparation method and imaging device, as well as an improved objective lens module, a combination of components and an external module, and an improved external imaging module and component combination are provided to solve at least one of the above-mentioned problems.
  • an imaging system including, in order from the object side to the image side along the optical axis: a light source configured to provide light irradiated to a sample; an imaging lens group configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the imaging system;
  • the plane where the phase modulation unit is located and the plane where the light source is located are a pair of conjugate planes; and at least the plane where the phase modulation unit is located and the imaging plane of the imaging system No lens or lens group for Fourier transform is introduced into the optical path.
  • an imaging system comprising, in order from the object side to the image side along the optical axis: a light source configured to provide light irradiated to a sample; an imaging lens group configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the imaging system;
  • the imaging system has a target plane conjugate with the plane where the light source is located, and the plane where the phase modulation unit is located is located between the back focal plane of the imaging lens group and the target plane; and at least the optical path between the plane where the phase modulation unit is located and the imaging surface of the imaging system does not introduce a lens or lens group for performing Fourier transform.
  • an imaging system which includes, in order from the object side to the image side along the optical axis: a light source configured to provide light for irradiating a sample; an imaging lens group configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the imaging system; wherein the plane where the phase modulation unit is located and the plane where the light source is located are a pair of conjugate planes; and the phase modulation unit is adjacent to the imaging surface of the imaging system.
  • an imaging system which includes, in order from the object side to the image side along the optical axis: a light source configured to provide light for irradiating a sample; an imaging lens group configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the imaging system; wherein the plane where the phase modulation unit is located and the plane where the light source is located are a pair of conjugate planes; and an intermediate lens group is arranged between the phase modulation unit and the imaging surface of the imaging system, and a rear focal plane of the intermediate lens group deviates from the imaging surface of the imaging system.
  • a method for preparing an imaging system comprising: providing a light source, an imaging lens group and a phase modulation unit; arranging the light source, the imaging lens group and the phase modulation unit in sequence along the optical axis of the imaging lens group; adjusting the position of the phase modulation unit so that the plane where the phase adjustment unit is located and the plane where the light source is located are a pair of conjugate planes; wherein at least a lens or lens group for performing Fourier transform is not introduced into the optical path between the plane where the phase modulation unit is located and the imaging plane of the imaging system.
  • an imaging device comprising the An imaging system and a photosensitive element, wherein the photosensitive surface of the photosensitive element coincides with the imaging surface of the imaging system.
  • an objective lens module comprising: a housing; an imaging lens group, disposed inside the housing, configured to receive light irradiated from a light source to a sample and emitted from the sample, so as to image the sample at least once; a phase modulation unit, disposed inside the housing, configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the objective lens module;
  • the plane where the phase modulation unit is located and the plane where the light source is located are a pair of conjugate planes, and at least the optical path between the plane where the phase modulation unit is located and the imaging plane of the objective lens module does not introduce a lens or lens group for Fourier transform.
  • a combination of components including: a light source configured to provide light for irradiating a sample; and an external module having a connecting portion, the connecting portion being connectable to an objective lens; wherein the external module further has a phase modulation unit;
  • the phase modulation unit is configured to modulate the light emitted through the objective lens to form a desired sample image on the imaging surface of the objective lens; wherein the plane where the phase modulation unit is located is conjugate with the plane where the light source is located, and at least the optical path between the plane where the phase modulation unit is located and the imaging surface of the objective lens does not introduce a lens or lens group that performs Fourier transform.
  • an external module which is applied to the combination as described above, and the external module includes a shell for accommodating the phase modulation unit, and the connecting part is provided on the shell.
  • an external imaging module comprising: a housing; an imaging lens group, disposed inside the housing, configured to receive light emitted by an objective lens, so as to perform a second or more sequential imaging of a sample; wherein the objective lens is configured to receive light irradiated from a light source to the sample and emitted by the sample, so as to perform a first imaging of the sample; a phase modulation unit, disposed inside the housing, configured to modulate the light emitted by the imaging lens group, so as to form a desired sample image on an imaging surface of the external imaging module;
  • the plane where the phase modulation unit is located and the plane where the light source is located are a pair of conjugate planes, and at least the optical path between the plane where the phase modulation unit is located and the imaging plane of the external imaging module does not introduce a lens or lens group for Fourier transform.
  • a combination of components is provided, which is applied to a microscope, comprising: a light source configured to provide light to illuminate a sample; and an external Imaging module.
  • FIG1 is a schematic diagram of an imaging optical path according to an embodiment of the present application.
  • FIG2 is a schematic diagram of an imaging optical path according to an embodiment of the present application.
  • FIG3 is a schematic diagram of component connections according to an embodiment of the present application.
  • FIG4 is a schematic diagram of adjusting a phase modulation unit for centering and eccentricity according to an embodiment of the present application
  • FIG5 is a schematic diagram of an imaging optical path according to an embodiment of the present application.
  • FIG6 is a schematic diagram of adjusting the front lens for centering and decentering according to an embodiment of the present application.
  • FIG7 is a schematic diagram of an imaging optical path according to an embodiment of the present application.
  • FIG8 is a 10-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG9 is a 20-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG10 is a 40-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG11 is a schematic structural diagram of an objective lens module according to an embodiment of the present application.
  • FIG12 is a schematic structural diagram of an objective lens module according to an embodiment of the present application.
  • FIG13 is a 10-fold magnification comparison schematic diagram of a tumor cell sample observed by an objective lens module according to an embodiment of the present application.
  • FIG14 is a 20-fold magnification schematic diagram of a tumor cell sample observed by an objective lens module according to an embodiment of the present application when the light source imaging coincides with the SPP center;
  • FIG15 is a schematic diagram of a 40-fold magnification of a tumor cell sample observed by an objective lens module according to an embodiment of the present application when the light source imaging coincides with the SPP center;
  • FIG16 is a 10-fold magnification comparison diagram of an unstained plant rhizome slice observed by an objective lens module according to an embodiment of the present application.
  • FIG17 is a 10-fold magnification comparison diagram of a diatom sample observed by an objective lens module according to an embodiment of the present application.
  • FIG18 is a schematic diagram of a combination of components according to an embodiment of the present application.
  • FIG19 is a schematic diagram of the combination of components and the coordination of an objective lens according to an embodiment of the present application.
  • FIG20 is a schematic diagram of the structure of an external module according to an embodiment of the present application.
  • FIG21 is a schematic diagram of the structure of an external imaging module according to an embodiment of the present application.
  • FIG22 is a schematic diagram of the structure of an external imaging module according to an embodiment of the present application.
  • FIG23 is a 10-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG24 is a 20-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG25 is a 50-fold magnified microscopic phase contrast imaging comparison diagram obtained in an embodiment of the present application.
  • FIG. 26 is a schematic structural diagram of a combination of components according to an embodiment of the present application.
  • Imaging system, 110, light source, 110'-110"' image of the light source, 120, imaging lens group, 130, phase modulation unit, 131, first position, 132, second position, 140, first housing, 150, second housing, 10A, object plane, 10B, imaging plane
  • 300 imaging system, 310, light source, 310', convergence point of the light source, 310", image of the convergence point of the light source, 310'", image of the convergence point of the light source after the position of the front lens is changed, 320, front lens, 330, imaging lens group, 340, phase modulation unit, 341, center of the phase modulation unit, 30A, object plane, 30B, imaging plane
  • first and second are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as “first” and “second” may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality” is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.
  • the space on one side of the object relative to the optical element is called the object side of the optical element, and correspondingly, the space on one side of the image of the object relative to the optical element is called the image side of the optical element.
  • the positional relationship of "adjacent" means that when the aperture is not considered, no other elements are arranged between the elements in the imaging system, between the elements and the object plane, and between the elements and the imaging plane.
  • the light source may include an entity that emits light itself, and may also include an entity (such as a reflector) that reflects incident light and causes the light to illuminate the sample.
  • the phase modulation unit may include one or more of a spiral phase plate (Spiral Phase Plate, SPP), a holographic grating, a mode converter including a spherical lens and a cylindrical lens, and a spatial light modulator.
  • the spiral phase plate is also called a spiral phase plate, or a vortex optical element, or a Bessel amplitude modulated spiral phase plate, which can convert the input Gaussian light into a circular energy ring, that is, to generate vortex light, and its structure is similar to the shape of a spiral or a spiral staircase.
  • the purpose of the design of this spiral is to control the phase of the vortex light beam.
  • the spiral phase plate is the simplest, most direct and most universal method to obtain vortex light.
  • Vortex beams can be generated, such as using a holographic grating to generate a vortex beam from low-order Gaussian light; a mode converter including a spherical lens and a cylindrical lens can also be used to obtain a vortex beam from high-order Hermitian Gaussian light; a spatial light modulator can also be selected to generate a vortex beam, etc.
  • phase contrast microscopes In order to achieve microscopic stereoscopic imaging of transparent samples, traditional phase contrast microscopes, Hoffman phase contrast microscopes, differential interference microscopes, etc. are usually used to highlight the phase information of transparent samples, and then observe transparent samples.
  • phase contrast (phase contrast) microscopes use the transmission ring in the light source and the dark field phase ring at the rear focal plane of the objective lens to convert phase information into amplitude information, thereby observing transparent samples;
  • Hoffman phase contrast microscopes use oblique incident light sources in combination with Hoffman grayscale filters to obtain three-dimensional morphological information of transparent samples;
  • differential interference microscopes use two beams of slightly offset incident light to irradiate the sample, carry the sample phase gradient information, and then integrate it into intensity information to present in the final image. Therefore, the position where the sample has a phase gradient will appear different from the light intensity distribution in the flat area, showing a relief-like effect.
  • phase contrast microscope is usually used to achieve edge enhancement imaging effect
  • Hoffman phase contrast microscope is usually used to achieve relief imaging effect
  • differential interference microscope is also usually used to achieve relief imaging effect.
  • Hoffman phase contrast microscope and differential interference microscope also have requirements for the observed samples. For example, when Hoffman microscope is used to shoot thicker samples, light and dark background stripes are easily formed, and differential interference microscope has requirements for birefringent optical path, which means that there must be no polarization-sensitive materials in the sample.
  • phase contrast microscope is usually used to achieve edge enhancement imaging effect
  • Hoffman phase contrast microscope is usually used to achieve relief imaging effect
  • differential interference microscope is also usually used to achieve relief imaging effect.
  • Hoffman phase contrast microscope and differential interference microscope also have requirements for the observed samples. For example, when Hoffman microscope is used to shoot thicker samples, light and dark background stripes are easily formed, and differential interference microscope has requirements for birefringent optical path, which means that there must be no polarization-sensitive materials in the sample.
  • the above constraints limit the application scope of traditional phase contrast microscopes.
  • the present application provides an imaging system, which places the phase modulation unit in a plane conjugate with the plane where the light source is located, thereby avoiding the addition of an additional lens for Fourier transform, thereby enriching the imaging effect of the phase contrast microscope while ensuring the imaging quality of the phase contrast microscope.
  • an embodiment of the present application provides an imaging system 100.
  • the imaging system 100 includes, in order from the object side to the image side along the optical axis AX1: a light source 110 configured to provide light to illuminate a sample; an imaging lens group 120 configured to receive light emitted from the sample to image the sample; and a phase modulation unit 130 configured to modulate the light emitted by the imaging lens group 120 to obtain a desired sample image on the imaging surface of the imaging system.
  • the plane where the phase modulation unit 130 is located and the plane where the light source 110 is located are a pair of conjugate planes; and at least the optical path between the plane where the phase modulation unit 130 is located and the imaging plane 10B of the imaging system 100 does not introduce a lens or lens group for Fourier transform.
  • FIG1 shows an imaging optical path of an imaging system 100.
  • the sample is located between the light source 110 and the imaging lens group 120, and its location is shown by the object plane 10A.
  • the light source 110 provides light to illuminate the sample, and after being emitted from the sample, light carrying sample information is formed. After being emitted from the imaging lens group 120, the light carrying sample information is emitted or reflected by the phase modulation unit 130, and finally reaches the imaging plane 10B of the imaging system 100.
  • the light source 110 includes a parallel light source and a point light source, and may also include a line light source or a surface light source that can be equivalent to a point light source.
  • the imaging lens group 120 includes at least one converging lens.
  • the imaging lens group 120 as a whole has a converging effect on light, or the imaging lens group 120 as a whole has a positive optical power.
  • the plane where the phase modulation unit 130 is located coincides with the back focal plane of the imaging lens group 120.
  • conjugation when an object is imaged by an optical system, the object point and the image point are in one-to-one correspondence, so the object point and the image point are a pair of conjugate points, and correspondingly, the plane where the object point is located and the plane where the image point is located are a pair of conjugate surfaces.
  • the image formed by the light source 110 through the imaging lens group 120 is the image point of the light source 110.
  • the image point of the light source 110 is located in the plane where the phase modulation unit 130 is located, it means that the plane where the phase modulation unit 130 is located and the plane where the light source 110 is located are a pair of conjugate surfaces.
  • the optical path between the plane where the phase modulation unit 130 is located and the imaging surface 10B of the imaging system 100 does not introduce a lens or lens group that performs Fourier transform can be expressed as, no lens or lens group is set in the optical path, or it can be expressed that although a lens or lens group is set in the optical path, each lens therein does not perform Fourier transform on the light, or the lens group as a whole does not perform Fourier transform on the light.
  • the optical path between the plane where the phase modulation unit 130 is located and the imaging surface 10B of the imaging system 100 does not introduce a lens or lens group that performs Fourier transform can be exemplarily expressed as the following two cases:
  • phase modulation unit 130 is adjacent to the imaging surface 10B, that is, No additional lens element is provided between the phase modulation unit 130 and the imaging plane 10B. In this case, the optical path between the phase modulation unit 130 and the imaging plane 10B will not introduce a lens or lens group for Fourier transform.
  • the light field distribution of the spherical wave emitted by a point light source when it is transmitted to the object plane is:
  • R is the distance from the light source 110 to the object plane 10A on the optical axis AX1, x0 and y0 are the spatial coordinates on the object plane 10A, A( x0 , y0 ) is the sample transmission function, i is the imaginary unit, k is the wave number, ⁇ is a constant that does not affect the light field distribution, and exp represents an exponential function with base e.
  • the light field distribution E 1 (x 1 , y 1 ) close to the imaging lens group 120 can be obtained.
  • the light field distribution of the object light wave i.e., the spherical wave carrying the object information
  • the light field distribution of the object light wave on the imaging surface can be obtained:
  • x2 and y2 are spatial coordinates on the plane where the phase modulation unit 130 is located
  • x3 and y3 are spatial coordinates on the imaging surface 10B
  • ⁇ and ⁇ are both constants
  • d1 represents the distance between the plane where the phase modulation unit 130 is located and the imaging lens group 120 on the optical axis AX1
  • d2 represents the distance between the plane where the phase modulation unit 130 is located and the imaging surface 10B on the optical axis AX1
  • f is the focal length of the imaging lens group 120
  • F represents Fourier transform
  • u and v represent the spectral coordinates of Fourier transform
  • H( x2 , y2 ) represents the transmission function of the phase modulation unit 130
  • ⁇ represents the wavelength.
  • E 3 (x 3 ,y 3 ) is basically the same as the expression of the final light field function for phase contrast imaging using the 4f system, with only one more quadratic phase factor that does not affect the light field intensity distribution. Therefore, it can be seen that the configuration of the present application can achieve a phase contrast imaging effect that is basically consistent with the 4f system without the additional introduction of a lens for Fourier transform. At the same time, since the introduction of lenses is reduced, it is beneficial to reduce optical distortion and improve imaging resolution.
  • the imaging system 100 introduces a phase modulation unit 130 into the imaging optical path, and makes the plane where the phase modulation unit 130 is located and the plane where the light source 110 is located a pair of conjugate planes, thereby facilitating more diverse imaging compared to bright field observation (i.e., a micrograph of a planar visual effect).
  • the resolution can be prevented from decreasing and the imaging quality can be improved.
  • the imaging system 100 further includes a first shell 140 for accommodating the imaging lens group 120 ; and a second shell 150 for accommodating the phase modulation unit 130 , wherein the first shell 140 and the second shell 150 are integrally formed or detachably connected.
  • the first shell 140 can be used to fix the imaging lens group 120
  • the second shell 150 is used to fix the phase modulation unit 130;
  • the imaging lens group 120 and the phase modulation unit 130 can be arranged in the same lens barrel, for example, they can be arranged together in the objective lens, which is beneficial to the modularization of the imaging system 100.
  • the objective lens can be directly replaced with the original objective lens when necessary; when the first shell 140 and the second shell 150 are detachably connected, it means that the imaging lens group 120 and the phase modulation unit 130 are respectively fixed to different shells, for example, the imaging lens group 120 is the lens group in the objective lens, and the shell containing the phase modulation unit 130 is used as an external module. Then, by assembling the shell containing the phase modulation unit 130 to the objective lens, the phase modulation unit 130 is located in a plane conjugate with the plane where the light source 110 is located. In this way, there is no need to modify the structure of the objective lens.
  • the imaging system 100 can be obtained by assembling simple external modules, which greatly reduces the preparation cost.
  • the phase modulation unit includes a spiral phase plate.
  • a spiral phase plate By controlling the relative position relationship between the imaging of the light source on the plane where the spiral phase plate is located and the center of the spiral phase plate, different phase contrast imaging effects can be obtained.
  • the transmission function of the spiral phase plate can be expressed as Wherein, circ represents the aperture function, exp represents the exponential function with base e, r is the radial coordinate, ⁇ is the angular coordinate, l is an arbitrary integer, and Rspp is the radius of the spiral phase plate.
  • a phase contrast microscopy image with edge enhancement can be obtained; when the imaging of the light source in the plane where the spiral phase plate is located deviates from the center of the spiral phase plate, and the center of the spiral phase plate is still located in the plane where the spiral phase plate is located, a phase contrast microscopy image with a relief effect can be obtained.
  • Figures 8, 9 and 10 show the phase contrast microscopy comparison images under 10x, 20x and 40x objective lens observations, respectively.
  • the detailed texture of the transparent sample is not obvious; when the imaging of the light source coincides with the center of the spiral phase plate (i.e., centered), the detailed edges of the transparent sample are enhanced; when the imaging of the light source is slightly eccentric to the center of the spiral phase plate, the details of the transparent sample show a light and dark distribution, and a three-dimensional relief imaging effect is apparent; when the imaging of the light source is offset from the center of the spiral phase plate, the details of the transparent sample show a light and dark distribution, and a three-dimensional relief imaging effect is apparent. When focusing, the details of transparent samples have obvious relief imaging effect.
  • the position of the spiral phase plate can be adjusted so that the center of the spiral phase plate (black solid circle) moves between a first position 131 and a second position 132, so as to achieve the coincidence or deviation between the center of the spiral phase plate and the image 110' of the light source.
  • the imaging system 100 also includes a first adjustment mechanism (not shown) connected to the spiral phase plate, and the first adjustment mechanism can adjust the position of the spiral phase plate based on user operation.
  • the first adjustment mechanism can adjust the spiral phase plate in a two-dimensional plane, for example, the spiral phase plate can be translated or rotated in the plane where it is located, for example, a manual adjustment method can be used, such as a threaded hole can be opened on the housing that fixes the spiral phase plate, and the spiral phase plate can be adjusted in two dimensions by the cooperation of the screw and the threaded hole, and for another example, an electric adjustment mechanism can be used, such as adjusting the two-dimensional position by driving a motor.
  • a manual adjustment method can be used, such as a threaded hole can be opened on the housing that fixes the spiral phase plate, and the spiral phase plate can be adjusted in two dimensions by the cooperation of the screw and the threaded hole
  • an electric adjustment mechanism can be used, such as adjusting the two-dimensional position by driving a motor.
  • the second type as shown in FIG6, when the center of the spiral phase plate is fixed at position 341, the imaging of the light source 310 in the plane where the spiral phase plate is located can be moved, so that the imaging of the light source coincides with or deviates from the center of the spiral phase plate.
  • the imaging system 300 also includes a front lens 320 located between the light source 310 and the imaging lens group 330, the front lens 320 is configured to converge the light emitted by the front lens 320 (the converged light is irradiated to the sample), and a second adjustment mechanism (not shown) connected to the front lens 320; wherein the second adjustment mechanism can change the position of the front lens 320 based on user operation, so that the imaging of the convergence point 310' of the light emitted by the light source 310 after the front lens 320 in the plane where the spiral phase plate (i.e., the phase modulation unit 340) is located coincides with the center of the spiral phase plate or deviates from the center of the spiral phase plate.
  • the spiral phase plate i.e., the phase modulation unit 340
  • the second adjustment mechanism controls the front lens 320 to be in the third position, the image of the convergence point 310' of the light source 310 is 310", and the image 310" coincides with the center of the spiral phase plate.
  • the second adjustment mechanism controls the front lens 320 to be in the fourth position, the image of the convergence point 310' of the light source 310 is 310"', and the image 310"' deviates from the center of the spiral phase plate.
  • the second adjustment mechanism can also adopt the aforementioned manual and electric adjustment mechanisms.
  • the third method is to synchronously change the position of the spiral phase plate and the position of the front lens.
  • a first adjustment mechanism and a second adjustment mechanism may be provided at the same time, so as to more conveniently achieve the coincidence or deviation of the imaging of the light source (or the light source convergence point) with the center of the spiral phase plate.
  • the imaging system 100 further includes a front lens 320 and an imaging lens.
  • the filter between the lens group 330 is configured to filter the light emitted by the front lens before the light is irradiated to the sample, so as to filter out the influence of ambient stray light on imaging and improve the imaging quality.
  • the imaging system 100 further includes a stage for carrying the sample, so that when the sample is thin, the distance from the object plane 10A of the imaging system 100 to the imaging lens group 120 can be expressed as the distance from the stage to the imaging lens group 120 on the optical axis; when the sample is thick, the distance from the object plane 10A of the imaging system 100 to the imaging lens group 120 also needs to consider the thickness of the sample, for example, it can be expressed by subtracting the sample thickness from the distance from the stage to the imaging lens group 120 on the optical axis.
  • the imaging lens group 420 includes, in sequence from the object side to the image side along the optical axis AX4: at least one imaging lens 421; and at least one relay lens 422; wherein the light carrying the sample information is sequentially emitted through at least one imaging lens 421 and at least one relay lens 422 to the phase modulation unit 430.
  • at least one relay lens 422 includes a first relay lens 4221 and a second relay lens 4222.
  • the plane where the phase modulation unit 130 is located coincides with the rear focal plane of the imaging lens group 120.
  • the plane where the phase modulation unit 130 is located i.e., the plane conjugated with the plane where the light source is located
  • the rear focal plane of the imaging lens group 120 are unified;
  • the position where the phase modulation unit 130 is placed along the optical axis AX1 has a certain tolerance, and is basically positioned between the rear focal plane of the imaging lens group 120 and the plane conjugated with the plane where the light source 110 is located.
  • using the rear focal plane of the imaging lens group 120 as the placement position of the phase modulation unit 130 can not only ensure the desired imaging effect, but also facilitate the preparation of the imaging system 100, and the above setting is applicable to most of the original microscopes, which is conducive to industrialization.
  • the imaging system 100 further includes a third adjustment mechanism (not shown), which is connected to the phase modulation unit 130 and is configured to change the position of the phase modulation unit 130 in the optical axis direction based on user operation.
  • a third adjustment mechanism (not shown), which is connected to the phase modulation unit 130 and is configured to change the position of the phase modulation unit 130 in the optical axis direction based on user operation.
  • the third adjustment mechanism There are three main reasons for setting the third adjustment mechanism. First, it can adapt to the illumination form of the point light source, so that the plane where the phase modulation unit 130 is located matches the plane where the imaging of the point light source 110 is located. Second, for imaging lens groups 120 with different magnifications, the positions of the rear focal planes are different.
  • the third adjustment mechanism it is convenient for the plane where the phase modulation unit 130 is located to match the rear focal planes of imaging lens groups 120 with different magnifications.
  • the position of the phase modulation unit 130 can be calibrated by the third adjustment mechanism.
  • the third adjustment mechanism may be a manual adjustment mechanism (such as It can be adjusted by a screw or gear transmission), or it can be an electric adjustment mechanism, such as adjusting by a drive motor.
  • an imaging system which includes, in order from the object side to the image side along the optical axis: a light source, configured to provide light for irradiating a sample; an imaging lens group, configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit, configured to modulate the light emitted from the imaging lens group to form a desired sample image on an imaging surface of the imaging system; wherein the imaging system has a target plane conjugate to the plane where the light source is located, and the plane where the phase modulation unit is located is located between the back focal plane of the imaging lens group and the target plane; and, at least the optical path between the plane where the phase modulation unit is located and the imaging surface of the imaging system does not introduce a lens or lens group that performs Fourier transform.
  • the phase modulation unit can be basically positioned between the rear focal plane of the imaging lens group and the target plane. At this time, compared with bright field observation, more diverse imaging effects can still be achieved, such as edge enhancement imaging effects and relief imaging effects. In addition, since at least the optical path between the plane where the phase modulation unit is located and the imaging surface of the imaging system does not introduce a lens or lens group for Fourier transform, the resolution can be avoided from being reduced and the imaging quality can be improved.
  • an imaging system 100 which includes, in order from the object side to the image side along the optical axis AX1: a light source 110, configured to provide light for irradiating a sample; an imaging lens group 120, configured to receive light emitted from the sample to image the sample at least once; and a phase modulation unit 130, configured to modulate the light emitted from the imaging lens group 120 to form a desired sample image on an imaging surface 10B of the imaging system 100; wherein the plane where the phase modulation unit 130 is located and the plane where the light source 110 is located are a pair of conjugate planes; and the phase modulation unit 130 is adjacent to the imaging surface 10B of the imaging system.
  • the imaging system 100 by introducing the phase modulation unit 130 into the imaging optical path, and making the plane where the phase modulation unit 130 is located and the plane where the light source 110 is located a pair of conjugate planes, is conducive to achieving more diverse imaging effects compared to bright field observation, such as edge enhancement imaging effects and relief imaging effects.
  • edge enhancement imaging effects and relief imaging effects since at least the optical path between the plane where the phase modulation unit 130 is located and the imaging surface 10B of the imaging system 100 does not introduce a lens or lens group for Fourier transform, it is possible to avoid resolution degradation and improve imaging quality.
  • the imaging system 200 includes, in order from the object side to the image side along the optical axis AX2: a light source 210 configured to provide light to illuminate the sample; an imaging lens group 220 configured to receive light emitted by the sample to perform imaging on the sample; At least one imaging is performed; and a phase modulation unit 230 is configured to modulate the light emitted by the imaging lens group 220 to form a desired sample image on the imaging surface 20B of the imaging system 200; wherein the plane where the phase modulation unit 230 is located and the plane where the light source 210 is located are a pair of conjugate planes; and an intermediate lens group 240 is arranged between the phase modulation unit 230 and the imaging surface 20B of the imaging system 200, and the back focal plane of the intermediate lens group 240 deviates from the imaging surface 20B of the imaging system 200.
  • the imaging system 200 by introducing the phase modulation unit 230 into the imaging optical path, and making the plane where the phase modulation unit 230 is located and the plane where the light source 210 is located a pair of conjugate planes, is conducive to achieving more diverse imaging effects compared to bright field observation, such as edge enhancement imaging effects and relief imaging effects.
  • edge enhancement imaging effects and relief imaging effects since at least the optical path between the plane where the phase modulation unit 230 is located and the imaging surface 20B of the imaging system 200 does not introduce a lens or lens group for Fourier transform, it is possible to avoid resolution degradation and improve imaging quality.
  • Another embodiment of the present application provides a method for preparing an imaging system, comprising:
  • the method for preparing the imaging system is to sequentially arrange the light source, the imaging lens group and the phase modulation unit, and adjust the phase modulation unit to a plane conjugate with the plane where the light source is located, thereby obtaining an imaging system that is conducive to achieving more diverse imaging effects.
  • the resolution can be avoided from being reduced and the imaging quality can be improved.
  • the plane where the adjusted phase modulation unit is located coincides with the rear focal plane of the imaging lens group. In this way, the required imaging effect can be ensured and the preparation of the imaging system can be facilitated.
  • an imaging device which includes the imaging system and a photosensitive element described in the above embodiments, wherein the photosensitive surface of the photosensitive element coincides with the imaging surface of the imaging system.
  • the photosensitive element may be a complementary metal oxide semiconductor (CMOS) image sensor or a charge-coupled device (CCD) image sensor.
  • CMOS complementary metal oxide semiconductor
  • CCD charge-coupled device
  • the above-mentioned imaging device can capture phase contrast microscopic images with different imaging effects by adjusting the relative position relationship between the phase modulation unit and the light source imaging, such as edge-enhanced phase contrast microscopic images and phase contrast microscopic images with relief effects. At the same time, the captured phase contrast microscopic images have higher imaging quality.
  • the present application also provides an objective lens module, which can be used with an eyepiece to facilitate users to observe transparent samples, or to take images of transparent samples through a camera after the objective lens module forms an image.
  • the objective lens module of the present application places the phase modulation unit on a plane conjugate with the plane where the light source is located, thereby avoiding the addition of an additional lens for Fourier transform, enriching the imaging effect of the phase contrast microscope without changing the original microscopic imaging optical path, and ensuring the imaging quality of the phase contrast microscope.
  • an embodiment of the present application provides an objective lens module 500, comprising: a housing 510; an imaging lens group 520, disposed inside the housing 510, configured to receive light emitted from the light source to the sample and emitted from the sample, so as to image the sample at least once; a phase modulation unit 530, disposed inside the housing 510, configured to modulate the light emitted from the imaging lens group 520 to form a desired sample image on the imaging surface of the objective lens module 500.
  • the plane where the phase modulation unit 530 is located and the plane where the light source is located are a pair of conjugate planes, and at least the optical path between the plane where the phase modulation unit 530 is located and the imaging surface of the objective lens module 500 does not introduce a lens or lens group for Fourier transform.
  • the imaging lens group 520 includes at least one converging lens.
  • the imaging lens group 520 as a whole has a converging effect on the light, or the imaging lens group 520 as a whole has a positive optical power.
  • the sample is located between the light source and the imaging lens group 520.
  • the light source provides light to irradiate the sample, which is emitted from the sample to form light carrying sample information.
  • the light carrying sample information is emitted from the imaging lens group 520, then emitted or reflected by the phase modulation unit 530, and finally reaches the imaging surface of the objective lens module 500.
  • conjugation when an object is imaged by an optical system, the object point and the image point are in one-to-one correspondence, so the object point and the image point are a pair of conjugate points, and correspondingly, the plane where the object point is located and the plane where the image point is located are a pair of conjugate surfaces.
  • the image formed by the light source through the imaging lens group 520 is the image point of the light source, and when the image point of the light source is located in the plane where the phase modulation unit 530 is located, it means that the plane where the phase modulation unit 530 is located and the plane where the light source is located are a pair of conjugate surfaces.
  • the optical path between the plane where the phase modulation unit 530 is located and the imaging surface of the objective lens module 500 does not introduce a lens or lens group for Fourier transform can be expressed as: No lens or lens group is provided, which may also mean that although a lens or lens group is provided in the optical path, each lens thereof does not perform Fourier transform on the light, or the lens group as a whole does not perform Fourier transform on the light.
  • the lens or lens group in the optical path has performed an effective Fourier transform.
  • the optical path between the plane where the phase modulation unit 530 is located and the imaging plane of the objective lens module 500 does not introduce a lens or lens group that performs Fourier transform can be exemplarily represented by the following two cases:
  • phase modulation unit 530 is adjacent to the imaging surface, that is, no other lens element is additionally arranged between the phase modulation unit 530 and the imaging surface, and at this time, the optical path between the phase modulation unit 530 and the imaging surface will definitely not introduce a lens or lens group for performing Fourier transform;
  • the rear focal plane ie, the Fourier transform plane
  • the objective lens module 500 introduces a phase modulation unit 530 into the microscopic imaging optical path, and makes the plane where the phase modulation unit 530 is located and the plane where the light source is located a pair of conjugate planes, thereby achieving more imaging effects, such as boundary enhancement imaging effects and relief imaging effects, without changing the original microscopic imaging optical path.
  • a lens or lens group for Fourier transform is not introduced into the optical path between the plane where the phase modulation unit 530 is located and the imaging surface of the objective lens module 500, the resolution can be avoided from being reduced and the imaging quality can be improved.
  • the housing 510 includes a light input end 513 and a light output end 514, and a light through hole 515 that passes through the light input end 513 and the light output end 514 along the optical axis AX5 of the objective lens module 500; wherein the imaging lens group 520 and the phase modulation unit 530 are both connected to the hole wall of the light through hole.
  • the phase modulation unit 530 can be integrated into the original objective lens to obtain a dedicated spiral phase contrast objective lens.
  • the spiral phase contrast objective lens can be directly used to replace the ordinary objective lens on the microscope to achieve effective observation of the transparent sample.
  • the imaging lens group 520 includes at least one imaging lens and at least one relay lens arranged in sequence from the object side to the image side along the optical axis AX5; wherein the light emitted from the sample is sequentially emitted from at least one imaging lens and at least one relay lens to the phase modulation unit 530.
  • the relay lens By setting the relay lens, it is beneficial to accurately position and better focus the sample, thereby improving the imaging clarity; at the same time, there is no need to be limited to the solution of adding a phase modulation unit to the original objective lens, and it can be The phase modulation unit is set by finding a plane position conjugate with the plane where the light source is located in the imaging optical path extended by the relay lens.
  • the objective lens module obtained by this embodiment can be applied to an inverted microscope, so that users can observe cultured living cells conveniently.
  • the housing 510 includes a first housing 511 and a second housing 512 that are detachably connected; wherein the imaging lens group 520 is disposed inside the first housing 511, and the phase modulation unit 530 is disposed inside the second housing 512.
  • the objective lens module 500 can be divided into at least two independent modules, which is convenient for cleaning, and when one of the modules is damaged, only the damaged module needs to be replaced, without replacing all the modules, thereby reducing maintenance costs.
  • the lens group in the ordinary objective lens can be used as the imaging lens group 520, that is, there is no need to modify the original objective lens (in other words, it is applicable to any original objective lens), and an external module provided with a phase modulation unit 530 can be connected to achieve an imaging effect that is basically consistent with the aforementioned spiral phase contrast objective lens, thereby eliminating the trouble of purchasing an additional spiral phase contrast objective lens or a mid-to-high-end phase contrast microscope, greatly reducing costs.
  • the plane where the phase modulation unit 530 is located coincides with the rear focal plane of the imaging lens group 520.
  • the plane where the phase modulation unit 530 is located i.e., the plane conjugated with the plane where the light source is located
  • the rear focal plane of the imaging lens group 520 are unified;
  • the phase modulation unit 530 has a certain tolerance for the position placed along the optical axis AX5 direction, and is basically positioned between the rear focal plane of the imaging lens group 520 and the plane conjugated with the plane where the light source is located.
  • the rear focal plane of the imaging lens group 520 is used as the placement position of the phase modulation unit 530, which can not only ensure the desired imaging effect, but also facilitate the preparation of the objective lens module 500, and the above setting is applicable to most of the original microscopes, which is conducive to industrialization.
  • the phase modulation unit 530 includes a spiral phase plate, so that when the imaging of the light source in the plane where the spiral phase plate is located coincides with the center of the spiral phase plate, an edge-enhanced phase contrast microscopy image can be obtained; when the imaging of the light source in the plane where the spiral phase plate is located deviates from the center of the spiral phase plate and the center of the spiral phase plate is still located in the plane where the spiral phase plate is located, a phase contrast microscopy image with a relief effect can be obtained.
  • Figures 13 to 15 respectively show a 10-fold magnification comparison diagram of a tumor cell sample observed by the objective lens module, a 20-fold magnification schematic diagram of a tumor cell sample observed when the light source imaging coincides with the SPP center, and a 40-fold magnification schematic diagram of a tumor cell sample observed when the light source imaging coincides with the SPP center.
  • Figure 16 shows a 10x magnification comparison of an unstained plant rhizome slice observed by the objective lens module. It can be seen that at the same magnification, when the image is observed in the bright field, the detailed texture of the transparent sample is not obvious, and the flat layer appears as a black network; when the light source imaging coincides with the center of the spiral phase plate, the detailed edge of the transparent sample is enhanced; when the light source imaging deviates from the center of the spiral phase plate, the details of the transparent sample are clearly contrasted with the background, forming a relief imaging effect.
  • FIG17 shows a 10-fold magnification comparison of a diatom sample observed by the objective lens module.
  • diatom samples since they have a certain thickness, the three-dimensional sense is stronger when observed using the objective lens module of this embodiment. It can be seen that at the same magnification, when the image is observed in the bright field, the detailed texture of the transparent sample is not obvious; when the light source imaging coincides with the center of the spiral phase plate, the detailed edge of the transparent sample is enhanced.
  • the objective lens module 500 may include a first adjustment mechanism (not shown).
  • the first adjustment mechanism is connected to the spiral phase plate, and is configured to change the position of the spiral phase plate in the plane where the spiral phase plate is located based on user operation, so that the center of the spiral phase plate coincides with or deviates from the imaging of the light source in the plane where the spiral phase plate is located.
  • the first adjustment mechanism can make the spiral phase plate translate or rotate in the plane where it is located.
  • it can be a manual adjustment mechanism, such as a threaded hole can be opened on the shell that fixes the spiral phase plate, and the two-dimensional position of the spiral phase plate can be adjusted by the cooperation of the screw and the threaded hole. It can also be an electric adjustment mechanism, such as adjustment by a driving motor.
  • a manual adjustment mechanism such as a threaded hole can be opened on the shell that fixes the spiral phase plate, and the two-dimensional position of the spiral phase plate can be adjusted by the cooperation of the screw and the threaded hole.
  • an electric adjustment mechanism such as adjustment by a driving motor.
  • the objective lens module 500 may further include a second adjustment mechanism (not shown).
  • the second adjustment mechanism is connected to the phase modulation unit 530, and is configured to change the position of the phase modulation unit 530 in the optical axis direction of the objective lens module 500 based on user operation. There are three main reasons for setting the second adjustment mechanism.
  • the second adjustment mechanism can be used to calibrate the position of the phase modulation unit 530 after the phase modulation unit 530 is offset.
  • the second adjustment mechanism can be a manual adjustment mechanism (such as adjustment through a screw or gear transmission) or an electric adjustment mechanism, such as adjustment through a drive motor.
  • the objective lens module 500 may also include the aforementioned first adjustment mechanism and the second adjustment mechanism at the same time, so as to better achieve the desired phase contrast imaging effect while ensuring the reliability of the objective lens module 500 .
  • another embodiment of the present application provides a combination of elements 600, including a light source 610, which is configured to provide light for irradiating a sample; and an external module 620, which has a connecting portion 621, and the connecting portion 621 can be connected to the objective lens; wherein the external module 620 also has a phase modulation unit 622; when the external module 620 is connected to the objective lens through the connecting portion 621, the phase modulation unit 622 is configured to modulate the light emitted by the objective lens to form a desired sample image on the imaging surface of the objective lens module having the objective lens; wherein the plane where the phase modulation unit 622 is located is conjugate with the plane where the light source 610 is located, and at least the optical path between the plane where the phase modulation unit 622 is located and the imaging surface of the objective lens module does not introduce a lens or lens group for Fourier transform.
  • the optical path does not introduce a lens or lens group for Fourier transform
  • the combination 600 of the above components can achieve an imaging effect that is basically the same as that of a spiral phase contrast objective lens by providing a light source 610 and an external module 620 with a phase modulation unit 622, and making the light source 610 illuminate the sample.
  • the external module 620 is connected to the light emitting end of the objective lens, and there is no need to modify the original objective lens, thereby eliminating the trouble of purchasing an additional spiral phase contrast objective lens or a mid-to-high-end phase contrast microscope, and greatly reducing the cost.
  • the light source 610 includes a parallel light source or a point light source.
  • the plane where the phase modulation unit 622 is located coincides with the back focal plane of the objective lens.
  • the imaging surface of the objective lens module may be an image surface of the sample image formed by observing the sample using the entire lens system including the objective lens. That is, the objective lens module may only be provided with an objective lens, or may be provided with other lenses in addition to the objective lens.
  • the phase modulation unit 622 includes a spiral phase plate
  • the external module 620 further includes a first adjustment mechanism (not shown), which is connected to the phase modulation unit 622 and is configured to change the position of the phase modulation unit 622 in the plane where the phase modulation unit 622 is located based on user operation, so that when the external module 620 is connected to the objective lens through the connection portion 621, the phase modulation unit The center of 622 coincides with or deviates from the imaging of the light source 610 in the plane where the phase modulation unit 622 is located.
  • the first adjustment mechanism when the imaging of the light source 610 in the plane where the phase modulation unit 622 is located coincides with the center of the phase modulation unit 622, an edge-enhanced phase-contrast microscopic image can be obtained; when the imaging of the light source 610 in the plane where the phase modulation unit 622 is located deviates from the center of the phase modulation unit 622, and the center of the phase modulation unit 622 is still located in the plane where the phase modulation unit 622 is located, a phase-contrast microscopic image with a relief effect can be obtained.
  • the specific setting form of the first adjustment mechanism can be referred to the aforementioned embodiment, and will not be repeated here.
  • the external module 620 further has a second adjustment mechanism (not shown), which is connected to the phase modulation unit and is configured to change the position of the phase modulation unit in the optical axis direction of the objective lens based on user operation.
  • a second adjustment mechanism (not shown), which is connected to the phase modulation unit and is configured to change the position of the phase modulation unit in the optical axis direction of the objective lens based on user operation.
  • the plane where the phase modulation unit 622 is located i.e., the plane conjugated with the plane where the light source is located
  • the back focal plane of the objective lens are unified.
  • the phase modulation unit 622 has a certain tolerance for the position placed along the optical axis direction, and is basically positioned between the back focal plane of the objective lens and the plane conjugated with the plane where the light source 610 is located. Therefore, as shown in FIG19 , when the external module 620 is connected to the objective lens through the connecting portion 621, the plane where the phase modulation unit 622 is located can be set to coincide with the back focal plane of the objective lens. In this way, the desired imaging effect can be ensured, and the preparation of the objective lens module 500 can be facilitated.
  • the above-mentioned setting is applicable to most of the original microscopes, which is conducive to industrialization.
  • the external module 700 includes a housing 710 for accommodating a phase modulation unit 720, and a connecting portion 711 that can be connected to an objective lens is disposed on the housing 710.
  • the plane where the phase modulation unit 720 is located can be conjugate with the plane where the light source is located, thereby achieving the desired phase contrast imaging effect.
  • the external module 700 can be directly assembled to the objective lens to obtain the desired phase contrast imaging effect.
  • the present application also provides an external imaging module that can be used with an objective lens to capture images of transparent samples through a camera.
  • the external imaging module of the present application places the phase modulation unit in a plane conjugate with the plane where the light source is located in the additional imaging optical path, thereby avoiding the need to add an additional lens for Fourier transform, enriching the imaging effect of the phase contrast microscope without changing the original microscope. Imaging optical path, and can ensure the imaging quality of phase contrast microscope.
  • an embodiment of the present application provides an external imaging module 800, comprising: a housing 810; an imaging lens group 820, disposed inside the housing 810, configured to receive light emitted through an objective lens, so as to image the sample for the second time or more than the second time; wherein the objective lens is configured to receive light emitted from the sample by the light source to image the sample for the first time; a phase modulation unit 830, disposed inside the housing 810, configured to modulate the light emitted through the imaging lens group 820 to form a desired sample image on the imaging surface of the external imaging module 800; wherein the plane where the phase modulation unit 830 is located and the plane where the light source is located are a pair of conjugate planes, and at least the optical path between the plane where the phase modulation unit 830 is located and the imaging surface of the external imaging module 800 does not introduce a lens or lens group for Fourier transform.
  • the imaging lens group 820 includes at least one converging lens.
  • the imaging lens group 820 includes at least one converging
  • the light emitted by the objective lens can be incident into the external imaging module 800 after the first imaging, and the external imaging module 800 will image the sample for the second time or more times, wherein the light carrying the sample information will be emitted by the imaging lens group 820, and then emitted or reflected by the phase modulation unit 830, and finally reach the imaging surface of the external imaging module 800.
  • the object point and the image point are in one-to-one correspondence, so the object point and the image point are a pair of conjugate points, and correspondingly, the plane where the object point is located and the plane where the image point is located are a pair of conjugate surfaces.
  • the image of the light source finally formed by the imaging lens group 820 is the image point of the light source.
  • the optical path between the plane where the phase modulation unit 830 is located and the imaging surface of the external imaging module 800 does not introduce a lens or lens group that performs Fourier transform can be expressed as no lens or lens group is set in the optical path, or it can also be expressed that although a lens or lens group is set in the optical path, each lens in it does not perform Fourier transform on the light, or the lens group as a whole does not perform Fourier transform on the light.
  • the light is Fourier transformed.
  • the lens or lens group in the light path has undergone an effective Fourier transform.
  • the light path between the plane where the phase modulation unit 830 is located and the imaging plane of the external imaging module 800 does not introduce a lens or lens group that performs Fourier transform can be exemplarily represented by the following two cases:
  • phase modulation unit 830 is adjacent to the imaging surface, that is, no other lens element is additionally arranged between the phase modulation unit 830 and the imaging surface, and at this time, the optical path between the phase modulation unit 830 and the imaging surface will definitely not introduce a lens or lens group for performing Fourier transform;
  • the rear focal plane ie, the Fourier transform plane
  • the above-mentioned external imaging module 800 can add an additional imaging light path after the original microscopic imaging light path, and set a phase modulation unit in the additional imaging light path, and make the plane where the phase modulation unit is located and the plane where the light source is located a pair of conjugate planes, so that more imaging effects, such as boundary enhancement imaging effect and relief imaging effect, can be achieved without changing the original microscopic imaging light path.
  • more imaging effects such as boundary enhancement imaging effect and relief imaging effect
  • at least the optical path between the plane where the phase modulation unit 830 is located and the imaging surface of the external imaging module does not introduce a lens or lens group for Fourier transform, it can avoid the adverse effects of reduced resolution and increased optical distortion caused by the introduction of additional lenses, thereby improving the imaging quality.
  • a connection portion 860 is provided on the housing 810, and the connection portion 860 can be connected to the photographic port of the microscope.
  • the photographic port of the microscope is originally the final imaging surface (i.e., the original imaging surface in FIG. 1 ).
  • the final imaging surface can be used as an intermediate image, projected to another distant imaging surface, and then captured by the camera.
  • the external imaging module 800 performs the last imaging of the sample.
  • the external imaging module 800 further includes a photosensitive element 850, and the photosensitive surface of the photosensitive element 850 coincides with the imaging surface of the external imaging module 800.
  • the photosensitive element can specifically be a complementary metal oxide semiconductor (CMOS) image sensor or a charge-coupled device (CCD) image sensor.
  • CMOS complementary metal oxide semiconductor
  • CCD charge-coupled device
  • the housing 810 is provided with a light-through hole 840.
  • the light hole 840 extends from one end of the housing 810 along the optical axis direction of the external imaging module 800, wherein the imaging lens group 820 and the phase modulation unit 830 are both connected to the hole wall of the light hole 840.
  • the imaging lens group 820 and the phase modulation unit 830 can be integrated into one module, thereby facilitating the disassembly and assembly of the external imaging module 800.
  • the housing 810 includes a first housing 811 and a second housing 812 that are detachably connected, wherein the imaging lens group 820 is disposed inside the first housing 811, and the phase modulation unit 830 is disposed inside the second housing 812.
  • the external imaging module 800 can be divided into at least two independent modules, thereby facilitating the cleaning of the external imaging module 800, and when one of the modules is damaged, only the damaged module needs to be replaced, without replacing all the modules, thereby reducing maintenance costs.
  • the plane where the phase modulation unit 830 is located coincides with the rear focal plane of the imaging lens group 820. Since the phase modulation unit 830 has a certain tolerance for the position placed along the optical axis, it is basically positioned between the rear focal plane of the imaging lens group 820 and the plane conjugate with the plane where the light source is located.
  • the rear focal plane of the imaging lens group 820 is used as the placement position of the phase modulation unit 830, which can not only ensure the required imaging effect, but also facilitate the preparation of the external imaging module 800, which is conducive to industrialization.
  • the phase modulation unit 830 includes a spiral phase plate, so that when the imaging of the light source in the plane where the spiral phase plate is located coincides with the center of the spiral phase plate, an edge-enhanced phase contrast microscopic image can be obtained; when the imaging of the light source in the plane where the spiral phase plate is located deviates from the center of the spiral phase plate, and the center of the spiral phase plate is still located in the plane where the spiral phase plate is located, a phase contrast microscopic image with a relief effect can be obtained.
  • Figures 23, 24 and 25 respectively show phase contrast microscopic comparison images of another transparent sample under 10x, 20x and 50x objective lenses.
  • the external imaging module 800 may include a first adjustment mechanism (not shown).
  • the first adjustment mechanism is connected to the spiral phase plate, and is configured to change the position of the spiral phase plate in the plane where the spiral phase plate is located based on user operation, so that the center of the spiral phase plate coincides with or deviates from the imaging of the light source in the plane where the spiral phase plate is located.
  • the first adjustment mechanism can make the spiral phase plate translate or rotate in the plane where it is located.
  • it can be a manual adjustment mechanism.
  • a threaded hole may be provided on the housing for fixing the spiral phase plate, and the two-dimensional position of the spiral phase plate may be adjusted by the cooperation between the screw and the threaded hole.
  • an electric adjustment mechanism may be provided, such as adjustment by a driving motor.
  • the external imaging module 800 may further include a second adjustment mechanism (not shown).
  • the second adjustment mechanism is connected to the phase modulation unit 830, and is configured to change the position of the phase modulation unit 830 in the optical axis direction of the external imaging module 800 based on user operation.
  • the second adjustment mechanism can be a manual adjustment mechanism (such as adjustment by a screw or gear transmission), or it can be an electric adjustment mechanism, such as adjustment by a drive motor.
  • the external imaging module 800 may also include the aforementioned first adjustment mechanism and the second adjustment mechanism at the same time to better achieve the desired phase contrast imaging effect while ensuring the reliability of the external imaging module 800 .
  • FIG. 26 another embodiment of the present application provides a combination of components 900 that can be applied to a microscope, including: a light source 910 configured to provide light to illuminate a sample; and an external imaging module 800 as described above.
  • the combination 900 of the above-mentioned components by providing a light source 910 and an external imaging module 800 equipped with a phase modulation unit 830, and making the light source 910 irradiate the sample, can achieve the desired phase contrast imaging effect through the external imaging module 800 without changing the original microscopic imaging optical path, and ensure the imaging quality, eliminating the trouble of purchasing additional mid-to-high-end phase contrast microscopes, and greatly reducing costs.
  • the numbers representing quantities or properties used to describe and claim certain embodiments of the present application should be understood to be modified by the terms “roughly”, “about”, “approximately” or “substantially” in some cases.
  • “roughly”, “about”, “approximately” or “substantially” can indicate a ⁇ 20% variation of the value it describes.
  • the numerical parameters used in the specification and claims are approximate values, which may change according to the desired characteristics of individual embodiments.
  • the numerical parameters should take into account the specified significant digits and adopt the general method of retaining digits.
  • the numerical domains and parameters used to confirm the breadth of their range in some embodiments of the present application are approximate values, in specific embodiments, such numerical values are set within a feasible range. Be as precise as possible.

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Abstract

本发明涉及一种成像系统、制备方法及取像装置。上述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经成像透镜组出射的光线进行调制以于成像系统的成像面形成期望的样品图像；其中，相位调制单元所在的平面与光源所在的平面为一对共轭面；并且，至少相位调制单元所在的平面与成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。上述成像系统，可在基本无需改变原有的显微成像光路的情况下实现更多样的成像效果，同时可避免分辨率下降，提升成像品质。

Description

成像系统、制备方法及取像装置 技术领域
本发明涉及光学成像技术领域，特别是涉及一种成像系统、制备方法及取像装置。
背景技术
为了实现生物等弱对比度透明样品的光学显微观察，样品的相位因子起到了关键作用。在显微领域，主流的有传统相衬显微镜、霍夫曼相差显微镜、微分干涉显微镜等来凸显样品的相位信息，以方便弱衬度样品的观测。
然而，上述几类显微镜成像效果有限，为了在一套成像系统中同时实现边缘增强的成像效果和浮雕成像效果，frey A.Davis等人于2000年提出了一种基于4f系统的螺旋相衬成像系统。该系统是利用一对共焦的透镜，在成像光路中使用带有螺旋相位的空间光调制器进行滤波，从而最终在成像面实现边界凸显或者浮雕像的成像效果。
但是，上述基于4f系统的螺旋相衬成像系统的成像分辨率不高，且存在一定的光学畸变，还需要对原有的成像光路进行调整，不易直接装置于商业成套光学成像系统中。
发明内容
根据本申请的各种实施例，提供一种改进的成像系统、制备方法及取像装置，以及一种改进的物镜模组、元件的组合及外置模组，以及一种改进的外置成像模组及元件的组合，以解决上述问题中的至少之一。
根据本申请的一个方面，提供了一种成像系统，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；
其中，所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，至少所述相位调制单元所在的平面与所述成像系统的成像面 之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的另一个方面，提供了一种成像系统，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；
其中，所述成像系统具有与所述光源所在的平面共轭的目标平面，所述相位调制单元所在的平面位于所述成像透镜组的后焦面与所述目标平面之间；并且，至少所述相位调制单元所在的平面与所述成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的又一个方面，提供了一种成像系统，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，所述相位调制单元与所述成像系统的成像面相邻。
根据本申请的又一个方面，提供了一种成像系统，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，所述相位调制单元与所述成像系统的成像面之间设置有中间透镜组，所述中间透镜组的后焦面与所述成像系统的成像面偏离。
根据本申请的又一个方面，提供了一种成像系统的制备方法，包括：提供光源、成像透镜组和相位调制单元；将所述光源、所述成像透镜组和所述相位调制单元沿所述成像透镜组的光轴依序设置；调整所述相位调制单元的位置，使所述相位调整单元所在的平面与所述光源所在的平面为一对共轭面；其中，至少所述相位调制单元所在的平面与所述成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的又一个方面，提供了一种取像装置，包括如前文所述的 成像系统以及感光元件，所述感光元件的感光表面与所述成像系统的成像面重合。
根据本申请的又一个方面，提供了一种物镜模组，包括：壳体；成像透镜组，设于所述壳体内部，被配置为接收光源照射至样品并经样品出射的光线，以对样品进行至少一次成像；相位调制单元，设于所述壳体内部，被配置为对经所述成像透镜组出射的光线进行调制以于所述物镜模组的成像面形成期望的样品图像；
其中，所述相位调制单元所在的平面与光源所在的平面为一对共轭面，并且，至少所述相位调制单元所在的平面和所述物镜模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的又一个方面，提供了一种元件的组合，包括：光源，被配置为提供照射至样品的光线；以及，外置模组，具有连接部，所述连接部可与物镜连接；其中，所述外置模组还具有相位调制单元；
当所述外置模组通过所述连接部连接至物镜时，所述相位调制单元被配置为对经物镜出射的光线进行调制以于物镜的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面共轭，并且，至少所述相位调制单元所在的平面和物镜的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的又一个方面，提供了一种外置模组，应用于如前文所述的组合，所述外置模组包括用于容纳所述相位调制单元的壳体，所述壳体上设置有所述连接部。
根据本申请的又一个方面，提供了一种外置成像模组，包括：壳体；成像透镜组，设于所述壳体内部，被配置为接收经物镜出射的光线，以对样品进行第二次或第二次以上次序的成像；其中，物镜被配置为接收光源照射至样品并经样品出射的光线，以对样品进行第一次成像；相位调制单元，设于所述壳体内部，被配置为对经所述成像透镜组出射的光线进行调制以于所述外置成像模组的成像面形成期望的样品图像；
其中，所述相位调制单元所在的平面与光源所在的平面为一对共轭面，并且，至少所述相位调制单元所在的平面和所述外置成像模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
根据本申请的又一个方面，提供了一种元件的组合，应用于显微镜，包括：光源，被配置为提供照射至样品的光线；以及，如前文所述的外置 成像模组。
本申请的一个或多个实施例的细节在下面的附图和描述中提出。本申请的其他特征、目的和优点将从说明书、附图以及权利要求书变得明显。
附图说明
为了更好地描述和说明这里公开的那些发明的实施例或示例，可以参考一幅或多幅附图。用于描述附图的附加细节或示例不应当被认为是对所公开的发明、目前描述的实施例或示例以及目前理解的这些发明的最佳模式中的任何一者的范围的限制。
图1为本申请一实施例的成像光路示意图；
图2为本申请一实施例的成像光路示意图；
图3为本申请一实施例的部件连接示意图；
图4为本申请一实施例调整相位调制单元进行对中和偏心的示意图；
图5为本申请一实施例的成像光路示意图；
图6为本申请一实施例调整前置透镜进行对中和偏心的示意图；
图7为本申请一实施例的成像光路示意图；
图8为本申请一实施例所成的10倍放大的显微相衬成像对比图；
图9为本申请一实施例所成的20倍放大的显微相衬成像对比图；
图10为本申请一实施例所成的40倍放大的显微相衬成像对比图；
图11为本申请一实施例物镜模组的结构示意图；
图12为本申请一实施例物镜模组的结构示意图；
图13为本申请一实施例物镜模组观测的肿瘤细胞样品的10倍放大对比示意图；
图14为本申请一实施例物镜模组在光源成像与SPP中心重合时观测的肿瘤细胞样品的20倍放大示意图；
图15为本申请一实施例物镜模组在光源成像与SPP中心重合时观测的肿瘤细胞样品的40倍放大示意图；
图16为本申请一实施例物镜模组观测的未染色植物根茎切片的10倍放大对比图；
图17为本申请一实施例物镜模组观测的硅藻样品的10倍放大对比图；
图18为本申请一实施例的元件的组合的示意图；
图19为本申请一实施例的元件的组合与物镜的配合示意图；
图20为本申请一实施例的外置模组的结构示意图；
图21为本申请一实施例外置成像模组的结构示意图；
图22为本申请一实施例外置成像模组的结构示意图；
图23为本申请一实施例所成的10倍放大的显微相衬成像对比图；
图24为本申请一实施例所成的20倍放大的显微相衬成像对比图；
图25为本申请一实施例所成的50倍放大的显微相衬成像对比图；
图26为本申请一实施例元件的组合的结构示意图。
元件标号说明：
100、成像系统，110、光源，110’～110”’、光源的像，120、成像透镜组，
130、相位调制单元，131、第一位置，132、第二位置，140、第一壳体，150、第二壳体，10A、物平面，10B、成像面；
200、成像系统，210、光源，220、成像透镜组，230、相位调制单元，
240、中间透镜组，241、中间透镜组的后焦面，20A、物平面，20B、成像面；
300、成像系统，310、光源，310’、光源的会聚点，310”、光源的会聚
点的像，310”’、前置透镜位置改变后的光源的会聚点的像，320、前置透镜，330、成像透镜组，340、相位调制单元，341、相位调制单元的中心，30A、物平面，30B、成像面；
400、成像系统，410、光源，420、成像透镜组，421、成像透镜，422、
中继透镜组，4221、第一中继透镜，4222、第二中继透镜，430、相位调制单元，40A、物平面，40B、成像面；
500、物镜模组，510、壳体，511、第一壳体，512、第二壳体，513、光
线射入端，514、光线射出端，515、通光孔，520、成像透镜组，530、相位调制单元；
600、元件的组合，610、光源，620、外置模组，621、连接部，622、相
位调制单元；
700、外置模组，710、壳体，711、连接部，720、相位调制单元；
800、外置成像模组，810、壳体，811、第一壳体，812、第二壳体，820、
成像透镜组，830、相位调制单元，840、通光孔，850、感光元件，860、连接部；
900、元件的组合，910、光源。
具体实施方式
为使本发明的技术方案和有益效果能够更加明显易懂，下面通过列举具体实施例的方式进行详细说明。其中，附图不一定是按比例绘制的，局部特征可以被放大或缩小，以更加清楚的显示局部特征的细节；除非另有定义，本申请所使用的技术和科学术语与本申请所属的技术领域中的技术和科学术语的含义相同。
此外，术语“第一”、“第二”仅用于描述目的，而不能理解为指示或暗示相对重要性或者隐含指明所指示的技术特征的数量。由此，限定有“第一”、“第二”的特征可以明示或者隐含地包括至少一个该特征。在本发明的描述中，“多个”的含义是至少两个，例如两个，三个等，除非另有明确具体的限定。
需要说明的是，当元件被称为“固定于”或“设置于”另一个元件，它可以直接在另一个元件上或者也可以存在居中的元件。当一个元件被认为是“连接”另一个元件，它可以是直接连接到另一个元件或者可能同时存在居中元件。本文所使用的术语“垂直的”、“水平的”、“上”、“下”、“左”、“右”以及类似的表述只是为了说明的目的，并不表示是唯一的实施方式。
在本说明书中，物体相对于光学元件所处的一侧空间称为该光学元件的物侧，对应的，物体所成的像相对于光学元件所处的一侧空间称为该光学元件的像侧。在本说明书中，“相邻”的位置关系表示在不考虑光阑的情形下，成像系统中元件与元件之间、元件与物平面之间、元件与成像面之间未设置其他元件。在本说明书中，光源可包括自身发出光线的实体，也可包括反射入射光线进而使光线照射到样品的实体(如反光镜)。
在本说明书中，相位调制单元可以包括螺旋相位板(Spiral Phase Plate，SPP)、全息光栅、包含球形透镜和柱透镜的模式转换器、空间光调制器中的一个或多个。其中，螺旋相位板也叫螺旋相位片，或涡旋光元件，或贝塞尔振幅调制螺旋相位片，可将输入的高斯光转换成圆环形能量环，也就是产生涡旋光，其结构类似于螺旋或螺旋阶梯的形状，这种螺旋的设计其目的是控制涡旋光束的相位。螺旋相位片是获得涡旋光最简单、直接、最通用的方法，它能够方便地设计涡旋光的直径和拓扑荷数，满足用户的实际需求。目前，除了可用螺旋相位板产生涡旋光束之外，还有许多方法 可以产生涡旋光束，如运用全息光栅，由低阶高斯光产生涡旋光束；也可采用包含球形透镜和柱透镜的模式转换器，由高阶厄米原高斯光获得涡旋光束；还可选择空间光调制器来产生涡旋光束等。
为实现透明样品的显微立体成像，通常可使用传统相衬显微镜、霍夫曼相差显微镜、微分干涉显微镜等来凸显透明样品的相位信息，进而对透明样品进行观测。具体而言，相衬(相差)显微镜基于阿贝成像原理，利用光源中的透射环与物镜后焦面的暗场相位环，把相位信息转化为振幅信息，从而观测透明样品；霍夫曼相差显微镜，利用斜入射光源与霍夫曼灰度滤波片搭配，获得透明样品的三维形貌信息；微分干涉显微镜，利用两束微小偏移的入射光，辐照样品后，携带样品相位梯度信息，而后整合为强度信息呈现在最终图像中，因此样品有相位梯度的位置，会显得不同于平直区域的光强分布，显现出类似浮雕的效果。
然而，上述相衬显微镜的成像效果有限，例如相衬显微镜通常用于实现边缘增强的成像效果，霍夫曼相差显微镜通常用于实现浮雕的成像效果，微分干涉显微镜也通常用于实现浮雕的成像效果。除此之外，霍夫曼相差显微镜和微分干涉显微镜还对观测样品有要求，例如霍夫曼显微镜用于拍摄较厚的样品时，容易形成明暗的背景条纹，而微分干涉显微镜对双折射光路有要求，意味着样品中不能有对偏振敏感的材质。上述约束都限制了传统相衬显微镜的应用范围。
另一方面，加载螺旋相位板的4f滤波系统被大量研究，以克服传统显微镜成像效果有限的问题。然而，这类系统需使用成对的透镜，也就是说，这类系统相较于本申请的方案会在成像过程中对物光进行傅里叶变换与逆傅里叶变换从而引入额外的透镜，进而导致成像分辨率下降以及光学畸变增加，较难兼顾成像品质。
至此，如何丰富相衬显微镜的成像效果同时保证相衬显微镜的成像品质，并尽量减少对原有光路的改动，成为应当予以考虑并解决的问题。
基于上述问题，本申请提供一种成像系统，将相位调制单元放置于与光源所在的平面共轭的平面，从而可避免加入额外的进行傅里叶变换的透镜，在丰富相衬显微镜的成像效果同时可保证相衬显微镜的成像品质。
如图1所示，本申请一实施例提供一种成像系统100。成像系统100沿着光轴AX1由物侧至像侧依序包括：光源110，被配置为提供照射至样品的光线；成像透镜组120，被配置为接收经样品出射的光线以对样品进 行至少一次成像；以及，相位调制单元130，被配置为对经成像透镜组120出射的光线进行调制以于成像系统的成像面得到期望的样品图像。
进一步的，相位调制单元130所在的平面与光源110所在的平面为一对共轭面；并且，至少相位调制单元130所在的平面与成像系统100的成像面10B之间的光路未引入进行傅里叶变换的透镜或透镜组。
图1示出了成像系统100的一种成像光路。在成像期间，样品位于光源110和成像透镜组120之间，其所在位置通过物平面10A示出，光源110提供照射至样品的光线，经样品出射后形成携带有样品信息的光线，携带有样品信息的光线经成像透镜组120出射后，再经相位调制单元130出射或反射，最终至成像系统100的成像面10B。
示例性的，光源110包括平行光源和点光源，也可以包括能够等效为点光源的线光源或面光源。示例性的，成像透镜组120包括至少一个会聚透镜。示例性的，成像透镜组120整体对光线起会聚作用，或者说成像透镜组120整体具有正光焦度。示例性的，光源110为平行光源时，相位调制单元130所在的平面与成像透镜组120的后焦面重合。
示例性的，关于共轭，当一个物体通过光学系统成像后，物点和像点是一一对应的，那么物点和和像点就是一对共轭点，对应的，物点所在的平面和像点所在的平面就是一对共轭面。从而，将光源110作为物点，那么光源110通过成像透镜组120所成的像就是光源110的像点，当光源110的像点位于相位调制单元130所在的平面时，表示相位调制单元130所在的平面与光源110所在的平面为一对共轭面。
示例性的，“相位调制单元130所在的平面与成像系统100的成像面10B之间的光路未引入进行傅里叶变换的透镜或透镜组”可表示为，该光路中没有设置透镜或透镜组，也可以表示该光路中虽然设置了透镜或透镜组，但其中的每个透镜均没有对光线进行傅里叶变换，或是透镜组整体没有对光线进行傅里叶变换。另一方面，透镜(或透镜组整体)的前后焦面上存在准确的傅里叶变换关系，从而，透镜(或透镜组整体)的后焦面可称为光路的傅里叶变换面，当透镜(或透镜组整体)的后焦面与成像面重合时，可称光路中的透镜或透镜组进行了有效的傅里叶变换。综上，“相位调制单元130所在的平面与成像系统100的成像面10B之间的光路未引入进行傅里叶变换的透镜或透镜组”可示例性地表示为以下两种情况：
(1)如图1所示，相位调制单元130与成像面10B相邻，也就是说， 相位调制单元130与成像面10B之间没有额外设置其他透镜元件，此时相位调制单元130与成像面10B之间的光路必然不会引入进行傅里叶变换的透镜或透镜组；
(2)如图2所示，相位调制单元230与成像面20B之间虽然设置有中间透镜组240，但是中间透镜组240的后焦面241(即傅里叶变换面)与成像面20B偏离。
下面将从成像系统100的成像过程来阐述成像系统100实现丰富相衬显微镜的成像效果的同时保证相衬显微镜的成像品质的原理：
在近轴近似下点光源发出的球面波传递至物平面时的光场分布为：
其中R为光源110到物平面10A的在光轴AX1上的距离，x0和y0为物平面10A上的空间坐标，A(x0,y0)为样品透射函数，i为虚数单位，k为波数，α为不影响光场分布的常数，exp表示以e为底的指数函数。
根据菲涅尔衍射公式，可得到紧靠成像透镜组120前的光场分布E1(x1,y1)，再经过成像透镜组120聚焦，可得到物光波(即携带了物体信息的球面波)传递至相位调制单元130所在表面时的光场分布为E2(x2,y2)，最终经滤波后可得到物光波在成像面的光场分布：
其中，x2和y2为相位调制单元130所在平面上的空间坐标，x3和y3为成像面10B上的空间坐标，γ和β均为常数，d1表示相位调制单元130所在的平面与成像透镜组120在光轴AX1上的距离，d2表示相位调制单元130所在的平面与成像面10B在光轴AX1上的距离，f为成像透镜组120的焦距，F表示傅里叶变换，u和v表示傅里叶变换的频谱坐标，H(x2,y2)表示相位调制单元130的透射函数，λ表示波长。
可以看到，E3(x3,y3)与利用4f系统实现相衬成像的最终光场函数的表达式基本相同，仅多出一项不影响光场强度分布的二次相位因子。因此，可知本申请的设置方式可以在不额外引入进行傅里叶变换的透镜的情况下，实现与4f系统基本一致的相衬成像效果，同时由于减少了透镜的引入，有利于减少光学畸变，提升成像分辨率。
上述成像系统100，通过在成像光路中引入相位调制单元130，并且使相位调制单元130所在的平面与光源110所在的平面为一对共轭面，从而相比于明场观测(即平面视觉效果的显微图)，有利于实现更多样的成像 效果，例如可以实现边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元130所在的平面与成像系统100的成像面10B之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
在一些实施方式中，如图3所示，成像系统100还包括用于容纳成像透镜组120的第一壳体140；以及用于容纳相位调制单元130的第二壳体150，其中，第一壳体140与第二壳体150一体成型或可拆卸连接。首先，第一壳体140可用于固定成像透镜组120，第二壳体150用于固定相位调制单元130；其次，当第一壳体140和第二壳体150一体成型时，表示成像透镜组120和相位调制单元130可设置在同一镜筒中，例如可一同设置在物镜中，从而有利于成像系统100的模块化，例如，需要时可直接用该物镜替换原本的物镜；当第一壳体140和第二壳体150可拆卸连接时，表示成像透镜组120和相位调制单元130分别固定于不同的壳体，例如成像透镜组120为物镜中的镜片组，而包含相位调制单元130的壳体则作为外部模块，接着，通过将包含相位调制单元130的壳体装配至物镜使相位调制单元130位于与光源110所在的平面共轭的平面，如此便可不用对物镜的结构进行改造，通过装配简单的外置模块即可得到成像系统100，大大降低了制备成本。
在一些实施方式中，相位调制单元包括螺旋相位板。通过控制光源于螺旋相位板所在平面的成像与螺旋相位板的中心的相对位置关系，可得到不同的相衬成像效果。示例性的，螺旋相位板的透射函数可表示为其中，circ表示孔径函数，exp表示以e为底的指数函数，r为径向坐标，θ为角向坐标，l为任意整数，Rspp为螺旋相位板半径。
示例性的，当光源于螺旋相位板所在平面的成像与螺旋相位板的中心重合时，可得到边缘增强的相衬显微图像；当光源于螺旋相位板所在平面的成像与螺旋相位板的中心偏离、且螺旋相位板的中心仍位于螺旋相位板所在的平面时，可得到浮雕效果的相衬显微图像。图8、图9和图10分别示出了10倍、20倍和40倍物镜观测下的相衬显微对比图像。可以看到，不同放大倍率下，当为明场观测图像时，透明样品的细节纹理并不明显；当光源成像与螺旋相位板的中心重合(即对中)时，透明样品的细节边缘得到增强；当光源成像与螺旋相位板的中心微偏心时，透明样品的细节呈现明暗分布，立体浮雕成像效果显现；当光源成像与螺旋相位板的中心偏 心时，透明样品的细节具备明显的浮雕成像效果。
下面将提供三种调整光源于螺旋相位板所在平面的成像与螺旋相位板的中心的相对位置关系的方式。
第一种，如图4所示，当光源110于螺旋相位板(即相位调制单元130)所在的平面的成像110’的位置固定，可调整螺旋相位板的位置，使螺旋相位板的中心(黑色实心圆)在一第一位置131和一第二位置132之间移动，实现螺旋相位板的中心与光源的成像110’的重合或偏离。示例性的，成像系统100还包括与螺旋相位板连接的第一调节机构(图未示出)，该第一调节机构可基于用户操作调整螺旋相位板的位置。示例性的，第一调节机构可以对螺旋相位板进行二维平面内的调节，例如可以使螺旋相位板在其所在的平面内平移或旋转，例如，可以采用手动调节方式，如在固定螺旋相位板的壳体上可开设螺纹孔，通过螺丝和螺纹孔的配合来对螺旋相位板进行二维位置的调节，又例如，可以采用电动调节机构，如通过驱动马达进行二维位置的调节。
第二种，如图6所示，当螺旋相位板的中心固定于位置341，可使光源310于螺旋相位板所在的平面的成像移动，实现光源的成像与螺旋相位板的中心重合或偏离。示例性的，如图5所示，成像系统300还包括位于光源310和成像透镜组330之间的前置透镜320，前置透镜320被配置为使经前置透镜320出射的光线会聚(会聚后的光线照射至样品)，以及与前置透镜320连接的第二调节机构(图未示出)；其中，第二调节机构可基于用户操作改变前置透镜320的位置，使光源310发出的光线经前置透镜320后的会聚点310’于螺旋相位板(即相位调制单元340)所在平面的成像与螺旋相位板的中心重合或偏离螺旋相位板的中心。因此，结合图5和图6可知，当第二调节机构控制前置透镜320处于第三位置时，光源310的会聚点310’的成像为310”，此时成像310”与螺旋相位板的中心重合，当第二调节机构控制前置透镜320处于第四位置时，光源310的会聚点310’的成像为310”’，此时成像310”’与螺旋相位板的中心偏离。示例性的，第二调节机构同样可采用前述手动和电动调节方式的机构。
第三种，可以同步改变螺旋相位板的位置和前置透镜的位置，例如，可以同时具备第一调节机构和第二调节机构，从而更方便地实现光源(或光源会聚点)的成像与螺旋相位板的中心重合或偏离。
在一些实施方式中，成像系统100还包括设于前置透镜320和成像透 镜组330之间的滤波片，被配置为在光线照射至样品前对经前置透镜出射的光线进行滤波，以滤除环境杂散光对成像的影响，提升成像品质。
在一些实施方式中，成像系统100还包括用于承载样品的载台，从而当样品较薄时，成像系统100的物平面10A至成像透镜组120的距离可表示为载台至成像透镜组120在光轴上的距离；当样品较厚时，则成像系统100的物平面10A至成像透镜组120的距离还需要考虑样品的厚度，例如可以用载台至成像透镜组120在光轴上的距离减去样品厚度来表示。
在一些实施方式中，如图7所示，成像透镜组420沿着光轴AX4由物侧至像侧依序包括：至少一个成像透镜421；以及至少一个中继透镜422；其中，携带有样品信息的光线依次经至少一个成像透镜421和至少一个中继透镜422出射后至相位调制单元430。示例性的，至少一个中继透镜422包括第一中继透镜4221和第二中继透镜4222。通过设置中继透镜，有利于精确定位和更好地聚焦样品，从而提高成像清晰度。如此，也有利于得到适用于倒置显微镜的物镜模组，从而方便用户对培养的活细胞的观测。
在一些实施方式中，继续参考图1，相位调制单元130所在的平面与成像透镜组120的后焦面重合。如此设置主要有两个方面的原因，一是原有显微光路中，大多采用平行光源入射，此时相位调制单元130所在的平面(即与光源所在的平面共轭的平面)和成像透镜组120的后焦面统一；二是相位调制单元130沿光轴AX1方向放置的位置具有一定的容忍度，基本定位于成像透镜组120的后焦面和与光源110所在的平面共轭的平面之间。综上，采用成像透镜组120的后焦面作为相位调制单元130的放置位置，既能保证所需的成像效果，也能方便成像系统100的制备，并且上述设置适用于原有的大部分显微镜，有助于实现产业化。
在一些实施方式中，继续参考图1，成像系统100还包括第三调节机构(图未示出)，与相位调制单元130连接，被配置为基于用户操作改变相位调制单元130在光轴方向上的位置。设置第三调节机构主要有三个方面的原因，一是可适应点光源的照射形式，方便相位调制单元130所在的平面匹配点光源110的成像所在的平面；二是对于不同放大倍率的成像透镜组120，其后焦面的位置不同，通过设置第三调节机构方便相位调制单元130所在的平面匹配不同放大倍率的成像透镜组120的后焦面；三是当相位调制单元130发生位置偏移后，可通过第三调节机构对相位调制单元130的位置进行校准。示例性的，第三调节机构可以是手动调节机构(如 通过丝杆或是齿轮传动调节)，也可以是电动调节机构，如通过驱动马达调节。
本申请另一实施例提供一种成像系统，成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经成像透镜组出射的光线进行调制以于成像系统的成像面形成期望的样品图像；其中，成像系统具有与光源所在的平面共轭的目标平面，相位调制单元所在的平面位于成像透镜组的后焦面与目标平面之间；并且，至少相位调制单元所在的平面与成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
上述成像系统，考虑到相位调制单元在光线传播方向放置的位置具有一定的容忍度，因此可将相位调制单元基本定位于成像透镜组的后焦面与目标平面之间，此时相比于明场观测，仍旧可实现更多样的成像效果，例如可以实现边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元所在的平面与成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
如图1所示，本申请又一实施例提供一种成像系统100，成像系统100沿着光轴AX1由物侧至像侧依序包括：光源110，被配置为提供照射至样品的光线；成像透镜组120，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元130，被配置为对经成像透镜组120出射的光线进行调制以于成像系统100的成像面10B形成期望的样品图像；其中，相位调制单元130所在的平面与光源110所在的平面为一对共轭面；并且，相位调制单元130与成像系统的成像面10B相邻。
上述成像系统100，通过在成像光路中引入相位调制单元130，并且使相位调制单元130所在的平面与光源110所在的平面为一对共轭面，从而相比于明场观测，有利于实现更多样的成像效果，例如可以实现边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元130所在的平面与成像系统100的成像面10B之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
如图2所示，本申请又一实施例提供一种成像系统200，成像系统200沿着光轴AX2由物侧至像侧依序包括：光源210，被配置为提供照射至样品的光线；成像透镜组220，被配置为接收经样品出射的光线以对样品进 行至少一次成像；以及，相位调制单元230，被配置为对经成像透镜组220出射的光线进行调制以于成像系统200的成像面20B形成期望的样品图像；其中，相位调制单元230所在的平面与光源210所在的平面为一对共轭面；并且，相位调制单元230与成像系统200的成像面20B之间设置有中间透镜组240，中间透镜组240的后焦面与成像系统200的成像面20B偏离。
上述成像系统200，通过在成像光路中引入相位调制单元230，并且使相位调制单元230所在的平面与光源210所在的平面为一对共轭面，从而相比于明场观测，有利于实现更多样的成像效果，例如可以实现边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元230所在的平面与成像系统200的成像面20B之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
本申请又一实施例提供一种成像系统的制备方法，包括：
S100、提供光源、成像透镜组和相位调制单元；
S200、将光源、成像透镜组和相位调制单元沿成像透镜组的光轴依序设置；
S300、调整相位调制单元的位置，使相位调整单元所在的平面与光源所在的平面为一对共轭面；其中，至少相位调制单元所在的平面与成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
上述成像系统的制备方法，通过依序排布光源、成像透镜组和相位调制单元，并将相位调制单元调整至与光源所在的平面共轭的平面，从而得到有利于实现更多样的成像效果的成像系统。除此之外，由于至少在相位调制单元所在的平面与成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
在一些实施方式中，调整后的相位调制单元所在的平面与成像透镜组的后焦面重合。如此，既能保证所需的成像效果，也能方便成像系统的制备。
本申请又一实施例提供一种取像装置，该取像装置包括前文各实施例所述的成像系统以及感光元件，感光元件的感光表面与成像系统的成像面重合。具体的，感光元件可以采用互补金属氧化物半导体(CMOS，Complementary Metal Oxide Semiconductor)图像传感器或者电荷耦合元件(CCD，Charge-coupled Device)图像传感器。
上述取像装置，通过调整相位调制单元与光源成像的相对位置关系，可拍摄得到不同成像效果的相衬显微图像，例如边缘增强的相衬显微图像和浮雕效果的相衬显微图像，同时，拍摄得到的相衬显微图像具备较高的成像品质。
本申请还提供一种物镜模组，可与目镜配合方便用户观测透明样品，或是在物镜模组成像后通过相机拍摄透明样品的图像。本申请的物镜模组将相位调制单元放置于与光源所在的平面共轭的平面，从而可避免加入额外的进行傅里叶变换的透镜，在丰富相衬显微镜的成像效果同时无需改变原有的显微成像光路，并可保证相衬显微镜的成像品质。
如图11所示，本申请一实施例提供一种物镜模组500，包括：壳体510；成像透镜组520，设于壳体510内部，被配置为接收光源照射至样品并经样品出射的光线，以对样品进行至少一次成像；相位调制单元530，设于壳体510内部，被配置为对经成像透镜组520出射的光线进行调制以于物镜模组500的成像面形成期望的样品图像。进一步的，相位调制单元530所在的平面与光源所在的平面为一对共轭面，并且，至少相位调制单元530所在的平面和物镜模组500的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。示例性的，成像透镜组520包括至少一个会聚透镜。示例性的，成像透镜组520整体对光线起会聚作用，或者说成像透镜组520整体具有正光焦度。
在成像期间，样品位于光源和成像透镜组520之间，光源提供照射至样品的光线，经样品出射后形成携带有样品信息的光线，携带有样品信息的光线经成像透镜组520出射后，再经相位调制单元530出射或反射，最终至物镜模组500的成像面。
示例性的，关于共轭，当一个物体通过光学系统成像后，物点和像点是一一对应的，那么物点和和像点就是一对共轭点，对应的，物点所在的平面和像点所在的平面就是一对共轭面。从而，将光源作为物点，那么光源通过成像透镜组520所成的像就是光源的像点，当光源的像点位于相位调制单元530所在的平面时，表示相位调制单元530所在的平面与光源所在的平面为一对共轭面。
示例性的，“相位调制单元530所在的平面与物镜模组500的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组”可表示为，该光路中 没有设置透镜或透镜组，也可以表示该光路中虽然设置了透镜或透镜组，但其中的每个透镜均没有对光线进行傅里叶变换，或是透镜组整体没有对光线进行傅里叶变换。另一方面，透镜(或透镜组整体)的前后焦面上存在准确的傅里叶变换关系，从而，透镜(或透镜组整体)的后焦面可称为光路的傅里叶变换面，当透镜(或透镜组整体)的后焦面与成像面重合时，可称光路中的透镜或透镜组进行了有效的傅里叶变换。综上，“相位调制单元530所在的平面与物镜模组500的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组”可示例性地表示为以下两种情况：
(1)如图11所示，相位调制单元530与成像面相邻，也就是说，相位调制单元530与成像面之间没有额外设置其他透镜元件，此时相位调制单元530与成像面之间的光路必然不会引入进行傅里叶变换的透镜或透镜组；
(2)相位调制单元530与成像面之间虽然设置有中间透镜组，但是中间透镜组的后焦面(即傅里叶变换面)与成像面偏离。
上述物镜模组500，通过在显微成像光路中引入相位调制单元530，并且使相位调制单元530所在的平面与光源所在的平面为一对共轭面，从而无需改变原有的显微成像光路便可实现更多成像效果，如边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元530所在的平面与物镜模组500的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免分辨率下降，提升成像品质。
在一些实施方式中，继续参考图11，壳体510包括光线射入端513和光线射出端514，以及沿物镜模组500的光轴AX5贯通光线射入端513和光线射出端514的通光孔515；其中，成像透镜组520与相位调制单元530均与通光孔的孔壁连接。通过上述方式，可将相位调制单元530集成在原有的物镜中从而得到专用的螺旋相差物镜，当用户观测透明样品时，可直接使用该螺旋相差物镜替换显微镜上的普通物镜，实现对透明样品的有效观测。
在一些实施方式中，成像透镜组520包括沿着光轴AX5由物侧至像侧依序设置的至少一个成像透镜和至少一个中继透镜；其中，经样品出射的光线依次经至少一个成像透镜和至少一个中继透镜出射后至相位调制单元530。通过设置中继透镜，有利于精确定位和更好地聚焦样品，从而提高成像清晰度；同时，无需限制于对原有物镜加装相位调制单元的方案，可在 由中继透镜拓展的成像光路中寻找与光源所在的平面共轭的平面位置来设置相位调制单元，换言之有更自由的位置选择空间来选择放置相位调制单元，进而得到本实施方式的物镜模组。通过本实施方式得到的物镜模组，可适用于倒置显微镜，从而方便用户对培养的活细胞进行观测。
在一些实施方式中，如图12所示，壳体510包括可拆卸连接的第一壳体511和第二壳体512；其中，成像透镜组520设于第一壳体511的内部，相位调制单元530设于第二壳体512的内部。如此，可将物镜模组500分为至少两个独立模块，从而方便清洗，并且，当其中一个模块损坏时，只需更换损坏的模块，无需更换全部模块，降低维护成本。除此之外，本实施方式中，可以将普通物镜中的镜片组作为成像透镜组520，即无需对原有的物镜进行改动(换言之适用于任何原有物镜)，通过外接一个设置有相位调制单元530的外置模组便可实现与前述螺旋相差物镜基本一致的成像效果，从而免去了额外购入螺旋相差物镜或是中高端相衬显微镜的麻烦，大大降低了成本。
在一些实施方式中，如图11所示，相位调制单元530所在的平面与成像透镜组520的后焦面重合。如此设置主要有两个方面的原因，一是原有显微光路中，大多采用平行光源照射样品，此时相位调制单元530所在的平面(即与光源所在的平面共轭的平面)和成像透镜组520的后焦面统一；二是相位调制单元530对沿光轴AX5方向放置的位置具有一定的容忍度，基本定位于成像透镜组520的后焦面和与光源所在的平面共轭的平面之间。综上，在物镜模组500中，采用成像透镜组520的后焦面作为相位调制单元530的放置位置，既能保证所需的成像效果，也能方便物镜模组500的制备，并且上述设置适用于原有的大部分显微镜，有助于实现产业化。
在一些实施方式中，相位调制单元530包括螺旋相位板，从而当光源于螺旋相位板所在平面的成像与螺旋相位板的中心重合时，可得到边缘增强的相衬显微图像；当光源于螺旋相位板所在平面的成像与螺旋相位板的中心偏离、且螺旋相位板的中心仍位于螺旋相位板所在的平面时，可得到浮雕效果的相衬显微图像。
示例性的，图13至图15分别示出了物镜模组观测的肿瘤细胞样品的10倍放大对比图、在光源成像与SPP中心重合时观测的肿瘤细胞样品的20倍放大示意图、在光源成像与SPP中心重合时观测的肿瘤细胞样品的40倍放大示意图。由图3可以看到，相同放大倍率下，当为明场观测图像 时(即平面视觉效果的显微图)，透明样品的细节纹理并不明显，细胞图像衬度较差；当光源成像与螺旋相位板的中心重合(即对中)时，透明样品的细节边缘得到增强；当光源成像与螺旋相位板的中心偏离(即偏心)时，透明样品的细节与背景对比明显，形成浮雕成像效果图。由图14和图15可以看到，不同放大倍率下，当光源成像与螺旋相位板的中心重合时，透明样品的细节边缘均得到加强。
示例性的，图16示出了物镜模组观测的未染色植物根茎切片的10倍放大对比图。可以看到，相同放大倍率下，当为明场观测图像时，透明样品的细节纹理并不明显，平层显黑色网络状；当光源成像与螺旋相位板的中心重合时，透明样品的细节边缘得到增强；当光源成像与螺旋相位板的中心偏离时，透明样品的细节与背景对比明显，形成浮雕成像效果。
示例性的，图17示出了物镜模组观测的硅藻样品的10倍放大对比图。对于硅藻类样品，因其本身具有一定厚度，使用本实施例的物镜模组观测，立体感更为强烈。可以看到，相同放大倍率下，当为明场观测图像时，透明样品的细节纹理并不明显；当光源成像与螺旋相位板的中心重合时，透明样品的细节边缘得到增强。
进一步的，为了实现上述光源于螺旋相位板(即相位调制单元530)所在平面的成像与螺旋相位板的中心的相对位置变化，物镜模组500可包括第一调节机构(图未示出)。具体的，第一调节机构与螺旋相位板连接，被配置为基于用户操作在螺旋相位板所在的平面内改变螺旋相位板的位置，使螺旋相位板的中心与光源于螺旋相位板所在的平面的成像重合或偏离光源于螺旋相位板所在的平面的成像。示例性的，第一调节机构可以使螺旋相位板在其所在的平面内平移或旋转，例如，可以是手动调节机构，如在固定螺旋相位板的壳体上可开设螺纹孔，通过螺丝和螺纹孔的配合来对螺旋相位板进行二维位置的调节，也可以是电动调节机构，如通过驱动马达调节。
在一些实施方式中，物镜模组500还可包括第二调节机构(图未示出)。具体的，第二调节机构与相位调制单元530连接，被配置为基于用户操作改变相位调制单元530在物镜模组500的光轴方向上的位置。设置第二调节机构主要有三个方面的原因，一是可适应点光源的照射形式，方便相位调制单元530所在的平面匹配点光源的成像所在的平面；二是对于不同放大倍率的成像透镜组520，其后焦面的位置不同，通过设置第二调节机构 方便相位调制单元530所在的平面匹配不同放大倍率的成像透镜组520的后焦面；三是当相位调制单元530发生位置偏移后，可通过第二调节机构对相位调制单元530的位置进行校准。示例性的，第二调节机构可以是手动调节机构(如通过丝杆或是齿轮传动调节)，也可以是电动调节机构，如通过驱动马达调节。
在一些实施方式中，物镜模组500也可同时包括前述第一调节机构和第二调节机构，以更好地实现期望的相衬成像效果，同时保证物镜模组500的可靠性。
如图18和图19所示，本申请另一实施例提供一种元件的组合600，包括光源610，被配置为提供照射至样品的光线；以及，外置模组620，具有连接部621，连接部621可与物镜连接；其中，外置模组620还具有相位调制单元622；当外置模组620通过连接部621连接至物镜时，相位调制单元622被配置为对经物镜出射的光线进行调制以于具有物镜的物镜模组的成像面形成期望的样品图像；其中，相位调制单元622所在的平面与光源610所在的平面共轭，并且，至少相位调制单元622所在的平面和物镜模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。其中，关于“光路未引入进行傅里叶变换的透镜或透镜组”可参见前述实施例中的阐释，此处不再赘述。
上述元件的组合600，通过提供光源610和设有相位调制单元622的外置模组620，并使光源610照射样品，外置模组620连接至物镜的光线射出端便可实现同螺旋相差物镜基本一致的成像效果，无需对原有的物镜进行改动，免去了额外购入螺旋相差物镜或是中高端相衬显微镜的麻烦，大大降低了成本。
示例性的，光源610包括平行光源或点光源。示例性的，光源610为平行光源时，相位调制单元622所在的平面与物镜的后焦面重合。
示例性的，物镜模组的成像面可以是利用包括物镜在内的整个透镜系统观测样品，形成样品图像的像面。也就是说，物镜模组中可以仅设有物镜，还可以设有除了物镜以外的其他透镜。
在一些实施方式中，相位调制单元622包括螺旋相位板，外置模组620还具有第一调节机构(图未示出)，与相位调制单元622连接，被配置为基于用户操作在相位调制单元622所在的平面内改变相位调制单元622的位置，以当外置模组620通过连接部621连接至物镜时，使相位调制单元 622的中心与光源610于相位调制单元622所在的平面的成像重合或偏离光源610于相位调制单元622所在的平面的成像。如此，通过第一调节机构，使得光源610于相位调制单元622所在平面的成像与相位调制单元622的中心重合时，可得到边缘增强的相衬显微图像；使得光源610于相位调制单元622所在平面的成像与相位调制单元622的中心偏离、且相位调制单元622的中心仍位于相位调制单元622所在的平面时，可得到浮雕效果的相衬显微图像。第一调节机构的具体设置形式可参见前述实施例，此处不再赘述。
在一些实施方式中，外置模组620还具有第二调节机构(图未示出)，与相位调制单元连接，被配置为基于用户操作改变相位调制单元在物镜的光轴方向上的位置。设置第二调节机构的原因以及第二调节机构的具体设置形式可参见前述实施例，此处不再赘述。
在一些实施方式中，由于原有显微光路中，大多采用平行光源照射样品，此时相位调制单元622所在的平面(即与光源所在的平面共轭的平面)和物镜的后焦面统一，另外，相位调制单元622对沿光轴方向放置的位置具有一定的容忍度，基本定位于物镜的后焦面和与光源610所在的平面共轭的平面之间。因此，如图19所示，当外置模组620通过连接部621连接至物镜时，可设置相位调制单元622所在的平面与物镜的后焦面重合，如此，既能保证所需的成像效果，也能方便物镜模组500的制备，并且上述设置适用于原有的大部分显微镜，有助于实现产业化。
如图20所示，本申请又一实施例提供一种外置模组700，可应用于前述元件的组合。外置模组700包括用于容纳相位调制单元720的壳体710，壳体710上设置有可与物镜连接的连接部711。
上述外置模组700，在装配至物镜时，可使相位调制单元720所在的平面与光源所在的平面共轭，从而实现所需的相衬成像效果。另一方面，对于已配置有光源的显微光路，可直接将外置模组700装配至物镜，进而得到所需的相衬成像效果。
本申请还提供一种外置成像模组，可与物镜配合通过相机拍摄透明样品的图像。本申请的外置成像模组是在额外增加的成像光路中将相位调制单元放置于与光源所在的平面共轭的平面，从而可避免加入额外的进行傅里叶变换的透镜，在丰富相衬显微镜的成像效果同时无需改变原有的显微 成像光路，并可保证相衬显微镜的成像品质。
如图21所示，本申请一实施例提供一种外置成像模组800，包括：壳体810；成像透镜组820，设于壳体810内部，被配置为接收经物镜出射的光线，以对样品进行第二次或第二次以上次序的成像；其中，物镜被配置为接收光源照射至样品并经样品出射的光线，以对样品进行第一次成像；相位调制单元830，设于壳体810内部，被配置为对经成像透镜组820出射的光线进行调制以于外置成像模组800的成像面形成期望的样品图像；其中，相位调制单元830所在的平面与光源所在的平面为一对共轭面，并且，至少相位调制单元830所在的平面和外置成像模组800的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。示例性的，成像透镜组820包括至少一个会聚透镜。示例性的，成像透镜组820整体对光线起会聚作用，或者说成像透镜组820整体具有正光焦度。
在成像期间，经物镜出射的光线在进行第一次成像后可入射进外置成像模组800，外置成像模组800将对样品进行第二次或第二次以上次序的成像，其中，携带有样品信息的光线经成像透镜组820出射后，将经相位调制单元830出射或反射，最终至外置成像模组800的成像面。具体的，“外置成像模组800将对样品进行第二次或第二次以上次序的成像”表示：当外置成像模组800与物镜直接邻接时，物镜与外置成像模组800之间的光路未引入其他透镜，此时外置成像模组800将对样品进行第二次成像；而当物镜与外置成像模组800之间的光路引入其他透镜时，其他透镜将对样品进行第二次成像，或是第二次、第三次成像，此时外置成像模组800将对样品进行第三次或是第四次成像。
示例性的，关于共轭，当一个物体通过光学系统成像后，物点和像点是一一对应的，那么物点和和像点就是一对共轭点，对应的，物点所在的平面和像点所在的平面就是一对共轭面。从而，将光源作为物点，那么光源最终通过成像透镜组820所成的像就是光源的像点，当光源的像点位于相位调制单元830所在的平面时，表示相位调制单元830所在的平面与光源所在的平面为一对共轭面。
示例性的，“相位调制单元830所在的平面与外置成像模组800的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组”可表示为，该光路中没有设置透镜或透镜组，也可以表示该光路中虽然设置了透镜或透镜组，但其中的每个透镜均没有对光线进行傅里叶变换，或是透镜组整体没有对 光线进行傅里叶变换。另一方面，透镜(或透镜组整体)的前后焦面上存在准确的傅里叶变换关系，从而，透镜(或透镜组整体)的后焦面可称为光路的傅里叶变换面，当透镜(或透镜组整体)的后焦面与成像面重合时，可称光路中的透镜或透镜组进行了有效的傅里叶变换。综上，“相位调制单元830所在的平面与外置成像模组800的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组”可示例性地表示为以下两种情况：
(1)如图1所示，相位调制单元830与成像面相邻，也就是说，相位调制单元830与成像面之间没有额外设置其他透镜元件，此时相位调制单元830与成像面之间的光路必然不会引入进行傅里叶变换的透镜或透镜组；
(2)相位调制单元830与成像面之间虽然设置有中间透镜组，但是中间透镜组的后焦面(即傅里叶变换面)与成像面偏离。
上述外置成像模组800，可在原有的显微成像光路之后增加额外的成像光路，并在该额外的成像光路中设置相位调制单元，且使相位调制单元所在的平面与光源所在的平面为一对共轭面，从而无需改变原有的显微成像光路便可实现更多成像效果，如边界增强的成像效果以及浮雕成像效果。除此之外，由于至少在相位调制单元830所在的平面与外置成像模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组，从而可避免引入额外透镜带来的分辨率下降、光学畸变增加等不利影响，提升成像品质。
在一些实施方式中，如图21所示，壳体810上设置有连接部860，连接部860可与显微镜的摄影口连接。继续参考图1，显微镜的摄影口本来为最终成像面(即图1中的原成像面)，在此处增加外置成像模组800，可将该最终成像面作为中间像，投射至另一远处的成像面，再被相机采集，此时，外置成像模组800对样品进行的是最后一次成像。通过在显微镜的摄影口设置新的成像光路，将不会对原有的显微成像光路造成影响。
在一些实施方式中，继续参考图21，外置成像模组800还包括感光元件850，感光元件850的感光表面与外置成像模组800的成像面重合。通过在外置成像模组800的成像面设置感光元件850，可拍摄透明样品的显微相衬图像，其中，感光元件具体可以采用互补金属氧化物半导体(CMOS，Complementary Metal Oxide Semiconductor)图像传感器或者电荷耦合元件(CCD，Charge-coupled Device)图像传感器。
在一些实施方式中，继续参考图21，壳体810开设有通光孔840，通 光孔840自壳体810的一端沿外置成像模组800的光轴方向延伸，其中，成像透镜组820与相位调制单元830均与通光孔840的孔壁连接。通过上述方式，可将成像透镜组820与相位调制单元830集成在一个模组中，从而方便外置成像模组800的拆装。
在一些实施方式中，如图22所示，壳体810包括可拆卸连接的第一壳体811和第二壳体812，其中，成像透镜组820设于第一壳体811的内部，相位调制单元830设于第二壳体812的内部。如此，可将外置成像模组800分为至少两个独立模块，从而方便外置成像模组800的清洗，并且，当其中一个模块损坏时，只需更换损坏的模块，无需更换全部模块，降低维护成本。
在一些实施方式中，相位调制单元830所在的平面与成像透镜组820的后焦面重合。由于相位调制单元830对沿光轴方向放置的位置具有一定的容忍度，基本定位于成像透镜组820的后焦面和与光源所在的平面共轭的平面之间。综上，在外置成像模组800中，采用成像透镜组820的后焦面作为相位调制单元830的放置位置，既能保证所需的成像效果，也能方便外置成像模组800的制备，有助于实现产业化。
在一些实施方式中，相位调制单元830包括螺旋相位板，从而当光源于螺旋相位板所在平面的成像与螺旋相位板的中心重合时，可得到边缘增强的相衬显微图像；当光源于螺旋相位板所在平面的成像与螺旋相位板的中心偏离、且螺旋相位板的中心仍位于螺旋相位板所在的平面时，可得到浮雕效果的相衬显微图像。图23、图24和图25分别示出了另一透明样品在10倍、20倍和50倍物镜观测下的相衬显微对比图像。可以看到，不同放大倍率下；当光源成像与螺旋相位板的中心重合(即对中)时，透明样品的细节边缘得到增强；当光源成像与螺旋相位板的中心偏心时，透明样品的细节具备明显的浮雕成像效果。
进一步的，为了实现上述光源于螺旋相位板(即相位调制单元830)所在平面的成像与螺旋相位板的中心的相对位置变化，外置成像模组800可包括第一调节机构(图未示出)。具体的，第一调节机构与螺旋相位板连接，被配置为基于用户操作在螺旋相位板所在的平面内改变螺旋相位板的位置，使螺旋相位板的中心与光源于螺旋相位板所在的平面的成像重合或偏离光源于螺旋相位板所在的平面的成像。示例性的，第一调节机构可以使螺旋相位板在其所在的平面内平移或旋转，例如，可以是手动调节机 构，如在固定螺旋相位板的壳体上可开设螺纹孔，通过螺丝和螺纹孔的配合来对螺旋相位板进行二维位置的调节，也可以是电动调节机构，如通过驱动马达调节。
在一些实施方式中，外置成像模组800还可包括第二调节机构(图未示出)。具体的，第二调节机构与相位调制单元830连接，被配置为基于用户操作改变相位调制单元830在外置成像模组800的光轴方向上的位置。设置第二调节机构主要由两个方面的原因，一是可适应点光源的照射形式，方便相位调制单元830所在的平面匹配点光源的成像所在的平面；二是当相位调制单元830发生位置偏移后，可通过第二调节机构对相位调制单元830的位置进行校准。示例性的，第二调节机构可以是手动调节机构(如通过丝杆或是齿轮传动调节)，也可以是电动调节机构，如通过驱动马达调节。
在一些实施方式中，外置成像模组800也可同时包括前述第一调节机构和第二调节机构，以更好地实现期望的相衬成像效果，同时保证外置成像模组800的可靠性。
如图26所示，本申请另一实施例提供一种元件的组合900，可应用于显微镜，包括：光源910，被配置为提供照射至样品的光线；以及，如前文所述的外置成像模组800。
上述元件的组合900，通过提供光源910和设有相位调制单元830的外置成像模组800，并使光源910照射样品，便可通过外置成像模组800实现期望的相衬成像效果，而无需对原有的显微成像光路进行改动，且保证了成像品质，免去了额外购入中高端相衬显微镜的麻烦，大大降低了成本。
需要指出的是，用于描述和要求保护本申请的某些实施例的表示数量或性质的数字应理解为在某些情况下被术语“大致”、“大约”、“近似”或“基本”修饰。例如，除非另外说明，否则“大致”、“大约”、“近似”或“基本”可以指示其所描述的值的±20％变化。相应地，在一些实施例中，说明书和权利要求中使用的数值参数均为近似值，该近似值根据个别实施例所需特点可以发生改变。在一些实施例中，数值参数应考虑规定的有效数位并采用一般位数保留的方法。尽管本申请一些实施例中用于确认其范围广度的数值域和参数为近似值，在具体实施例中，此类数值的设定在可行范围内 尽可能精确。
应当理解，以上实施例均为示例性的，不用于包含权利要求所包含的所有可能的实施方式。在不脱离本公开的范围的情况下，还可以在以上实施例的基础上做出各种变形和改变。同样的，也可以对以上实施例的各个技术特征进行任意组合，以形成可能没有被明确描述的本发明的另外的实施例。因此，上述实施例仅表达了本发明的几种实施方式，不对本发明专利的保护范围进行限制。

Claims (40)

  1. 一种成像系统，其特征在于，所述成像系统沿着光轴由物侧至像侧依序包括：
    光源，被配置为提供照射至样品的光线；
    成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，
    相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；
    其中，
    所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，至少所述相位调制单元所在的平面与所述成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  2. 根据权利要求1所述的成像系统，其特征在于，所述光源包括平行光源或点光源。
  3. 根据权利要求1所述的成像系统，其特征在于，所述相位调制单元与所述成像系统的成像面相邻。
  4. 根据权利要求1所述的成像系统，其特征在于，还包括：
    用于容纳所述成像透镜组的第一壳体；以及，
    用于容纳所述相位调制单元的第二壳体；
    其中，所述第一壳体与所述第二壳体一体成型或可拆卸连接。
  5. 根据权利要求1～4中任一项所述的成像系统，其特征在于，所述相位调制单元包括螺旋相位板。
  6. 根据权利要求5所述的成像系统，其特征在于，还包括：
    第一调节机构，与所述相位调制单元连接；
    其中，所述第一调节机构被配置为基于用户操作在所述相位调制单元所在的平面内改变所述相位调制单元的位置，使所述相位调制单元的中心与所述光源于所述相位调制单元所在的平面的成像重合或偏离所述光源于所述相位调制单元所在的平面的成像。
  7. 根据权利要求5所述的成像系统，其特征在于，还包括：
    前置透镜，位于所述光源和所述成像透镜组之间，被配置为使经所述前置透镜出射的光线会聚；以及，
    第二调节机构，与所述前置透镜连接；
    其中，所述第二调节机构被配置为基于用户操作改变所述前置透镜的位置，使所述光源发出的光线经所述前置透镜后的会聚点于所述相位调制单元所在平面的成像与所述相位调制单元的中心重合或偏离所述相位调制单元的中心。
  8. 根据权利要求7所述的成像系统，其特征在于，还包括：
    滤波片，设于所述前置透镜和所述成像透镜组之间，被配置为在光线照射至样品前对经所述前置透镜出射的光线进行滤波。
  9. 根据权利要求1～4中任一项所述的成像系统，其特征在于，所述成像透镜组沿着光轴由物侧至像侧依序包括：
    至少一个成像透镜；以及，
    至少一个中继透镜；
    其中，携带有样品信息的光线依次经所述至少一个成像透镜和所述至少一个中继透镜出射后至所述相位调制单元。
  10. 根据权利要求1～4中任一项所述的成像系统，其特征在于，所述相位调制单元所在的平面与所述成像透镜组的后焦面重合。
  11. 根据权利要求1～4中任一项所述的成像系统，其特征在于，所述成像系统还包括：
    第三调节机构，与所述相位调制单元连接，被配置为基于用户操作改变所述相位调制单元在光轴方向上的位置。
  12. 一种成像系统，其特征在于，所述成像系统沿着光轴由物侧至像侧依序包括：
    光源，被配置为提供照射至样品的光线；
    成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，
    相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；
    其中，所述成像系统具有与所述光源所在的平面共轭的目标平面，所述相位调制单元所在的平面位于所述成像透镜组的后焦面与所述目标平面之间；并且，至少所述相位调制单元所在的平面与所述成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  13. 一种成像系统，其特征在于，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，所述相位调制单元与所述成像系统的成像面相邻。
  14. 一种成像系统，其特征在于，所述成像系统沿着光轴由物侧至像侧依序包括：光源，被配置为提供照射至样品的光线；成像透镜组，被配置为接收经样品出射的光线以对样品进行至少一次成像；以及，相位调制单元，被配置为对经所述成像透镜组出射的光线进行调制以于所述成像系统的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面为一对共轭面；并且，所述相位调制单元与所述成像系统的成像面之间设置有中间透镜组，所述中间透镜组的后焦面与所述成像系统的成像面偏离。
  15. 一种成像系统的制备方法，其特征在于，包括：
    提供光源、成像透镜组和相位调制单元；
    将所述光源、所述成像透镜组和所述相位调制单元沿所述成像透镜组的光轴依序设置；
    调整所述相位调制单元的位置，使所述相位调整单元所在的平面与所述光源所在的平面为一对共轭面；
    其中，至少所述相位调制单元所在的平面与所述成像系统的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  16. 根据权利要求15所述的制备方法，其特征在于，调整后的所述相位调制单元所在的平面与所述成像透镜组的后焦面重合。
  17. 一种取像装置，其特征在于，包括如权利要求1～14中任一项所述的成像系统以及感光元件，所述感光元件的感光表面与所述成像系统的成像面重合。
  18. 一种物镜模组，其特征在于，包括：
    壳体；
    成像透镜组，设于所述壳体内部，被配置为接收光源照射至样品并经样品出射的光线，以对样品进行至少一次成像；
    相位调制单元，设于所述壳体内部，被配置为对经所述成像透镜组出射的光线进行调制以于所述物镜模组的成像面形成期望的样品图像；
    其中，
    所述相位调制单元所在的平面与光源所在的平面为一对共轭面，并且，至少所述相位调制单元所在的平面和所述物镜模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  19. 根据权利要求18所述的物镜模组，其特征在于，
    所述壳体包括光线射入端和光线射出端，以及沿所述物镜模组的光轴贯通所述光线射入端和所述光线射出端的通光孔；
    其中，
    所述成像透镜组与所述相位调制单元均与所述通光孔的孔壁连接。
  20. 根据权利要求19所述的物镜模组，其特征在于，
    所述成像透镜组包括沿着所述光轴由物侧至像侧依序设置的至少一个成像透镜和至少一个中继透镜；
    其中，经样品出射的光线依次经所述至少一个成像透镜和所述至少一个中继透镜出射后至所述相位调制单元。
  21. 根据权利要求18所述的物镜模组，其特征在于，
    所述壳体包括可拆卸连接的第一壳体和第二壳体；
    其中，
    所述成像透镜组设于所述第一壳体的内部，所述相位调制单元设于所述第二壳体的内部。
  22. 根据权利要求18～21中任一项所述的物镜模组，其特征在于，所 述相位调制单元所在的平面与所述成像透镜组的后焦面重合。
  23. 根据权利要求18～21中任一项所述的物镜模组，其特征在于，所述相位调制单元包括螺旋相位板。
  24. 根据权利要求23所述的物镜模组，其特征在于，还包括：
    第一调节机构，与所述相位调制单元连接，被配置为基于用户操作在所述相位调制单元所在的平面内改变所述相位调制单元的位置，使所述相位调制单元的中心与光源于所述相位调制单元所在的平面的成像重合或偏离光源于所述相位调制单元所在的平面的成像。
  25. 根据权利要求18～21中任一项所述的物镜模组，其特征在于，还包括：
    第二调节机构，与所述相位调制单元连接，被配置为基于用户操作改变所述相位调制单元在所述物镜模组的光轴方向上的位置。
  26. 一种元件的组合，其特征在于，包括：
    光源，被配置为提供照射至样品的光线；以及，
    外置模组，具有连接部，所述连接部可与物镜连接；
    其中，所述外置模组还具有相位调制单元；
    当所述外置模组通过所述连接部连接至物镜时，所述相位调制单元被配置为对经物镜出射的光线进行调制以于具有所述物镜的物镜模组的成像面形成期望的样品图像；其中，所述相位调制单元所在的平面与所述光源所在的平面共轭，并且，至少所述相位调制单元所在的平面和所述物镜模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  27. 根据权利要求26所述的组合，其特征在于，所述相位调制单元包括螺旋相位板，所述外置模组还具有：
    第一调节机构，与所述相位调制单元连接，被配置为基于用户操作在所述相位调制单元所在的平面内改变所述相位调制单元的位置，以当所述外置模组通过所述连接部连接至物镜时，使所述相位调制单元的中心与所述光源于所述相位调制单元所在的平面的成像重合或偏离所述光源于所述相位调制单元所在的平面的成像。
  28. 根据权利要求26或27所述的组合，其特征在于，所述外置模组 还具有：
    第二调节机构，与所述相位调制单元连接，被配置为基于用户操作改变所述相位调制单元在物镜的光轴方向上的位置。
  29. 根据权利要求26或27所述的组合，其特征在于，当所述外置模组通过所述连接部连接至物镜时，所述相位调制单元所在的平面与物镜的后焦面重合。
  30. 一种外置模组，其特征在于，应用于如权利要求26～29中任一项所述的组合，所述外置模组包括用于容纳所述相位调制单元的壳体，所述壳体上设置有所述连接部。
  31. 一种外置成像模组，其特征在于，包括：
    壳体；
    成像透镜组，设于所述壳体内部，被配置为接收经物镜出射的光线，以对样品进行第二次或第二次以上次序的成像；其中，物镜被配置为接收光源照射至样品并经样品出射的光线，以对样品进行第一次成像；
    相位调制单元，设于所述壳体内部，被配置为对经所述成像透镜组出射的光线进行调制以于所述外置成像模组的成像面形成期望的样品图像；
    其中，
    所述相位调制单元所在的平面与光源所在的平面为一对共轭面，并且，至少所述相位调制单元所在的平面和所述外置成像模组的成像面之间的光路未引入进行傅里叶变换的透镜或透镜组。
  32. 根据权利要求31所述的外置成像模组，其特征在于，所述壳体上设置有连接部，所述连接部可与显微镜的摄影口连接。
  33. 根据权利要求32所述的外置成像模组，其特征在于，还包括感光元件，所述感光元件的感光表面与所述外置成像模组的成像面重合。
  34. 根据权利要求31所述的外置成像模组，其特征在于，
    所述壳体开设有通光孔，所述通光孔自所述壳体的一端沿所述外置成像模组的光轴方向延伸，其中，所述成像透镜组与所述相位调制单元均与所述通光孔的孔壁连接。
  35. 根据权利要求31所述的外置成像模组，其特征在于，所述壳体包 括可拆卸连接的第一壳体和第二壳体，其中，所述成像透镜组设于所述第一壳体的内部，所述相位调制单元设于所述第二壳体的内部。
  36. 根据权利要求31～35中任一项所述的外置成像模组，其特征在于，所述相位调制单元所在的平面与所述成像透镜组的后焦面重合。
  37. 根据权利要求31～35中任一项所述的外置成像模组，其特征在于，所述相位调制单元包括螺旋相位板。
  38. 根据权利要求37所述的外置成像模组，其特征在于，还包括：
    第一调节机构，与所述相位调制单元连接，被配置为基于用户操作在所述相位调制单元所在的平面内改变所述相位调制单元的位置，使所述相位调制单元的中心与光源于所述相位调制单元所在的平面的成像重合或偏离光源于所述相位调制单元所在的平面的成像。
  39. 根据权利要求31～35中任一项所述的外置成像模组，其特征在于，还包括：
    第二调节机构，与所述相位调制单元连接，被配置为基于用户操作改变所述相位调制单元在所述外置成像模组的光轴方向上的位置。
  40. 一种元件的组合，其特征在于，应用于显微镜，包括：光源，被配置为提供照射至样品的光线；以及，如权利要求31～39中任一项所述的外置成像模组。
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