WO2019159627A1 - オートフォーカス装置ならびにそれを備える光学装置および顕微鏡 - Google Patents

オートフォーカス装置ならびにそれを備える光学装置および顕微鏡 Download PDF

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Publication number
WO2019159627A1
WO2019159627A1 PCT/JP2019/002216 JP2019002216W WO2019159627A1 WO 2019159627 A1 WO2019159627 A1 WO 2019159627A1 JP 2019002216 W JP2019002216 W JP 2019002216W WO 2019159627 A1 WO2019159627 A1 WO 2019159627A1
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Prior art keywords
light
optical system
image
light source
autofocus
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Ceased
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PCT/JP2019/002216
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English (en)
French (fr)
Japanese (ja)
Inventor
真人 安井
通夫 廣島
昌宏 上田
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RIKEN
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RIKEN
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Priority to CN201980013041.5A priority Critical patent/CN111819485B/zh
Priority to US16/967,814 priority patent/US11567293B2/en
Priority to DE112019000783.1T priority patent/DE112019000783T5/de
Priority to JP2020500356A priority patent/JP7226825B2/ja
Publication of WO2019159627A1 publication Critical patent/WO2019159627A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B7/00—Mountings, adjusting means, or light-tight connections, for optical elements
    • G02B7/28—Systems for automatic generation of focusing signals
    • G02B7/30—Systems for automatic generation of focusing signals using parallactic triangle with a base line
    • G02B7/32—Systems for automatic generation of focusing signals using parallactic triangle with a base line using active means, e.g. light emitter
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/241—Devices for focusing
    • G02B21/244—Devices for focusing using image analysis techniques
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/24—Base structure
    • G02B21/26—Stages; Adjusting means therefor
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03B—APPARATUS OR ARRANGEMENTS FOR TAKING PHOTOGRAPHS OR FOR PROJECTING OR VIEWING THEM; APPARATUS OR ARRANGEMENTS EMPLOYING ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ACCESSORIES THEREFOR
    • G03B13/00—Viewfinders; Focusing aids for cameras; Means for focusing for cameras; Autofocus systems for cameras
    • G03B13/32—Means for focusing
    • G03B13/34—Power focusing
    • G03B13/36—Autofocus systems
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/50—Constructional details
    • H04N23/55—Optical parts specially adapted for electronic image sensors; Mounting thereof
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/60—Control of cameras or camera modules

Definitions

  • the present disclosure relates to an autofocus device, an optical device including the autofocus device, and a microscope.
  • NA numerical aperture
  • FIG. 30 is a diagram for explaining a conventional first method.
  • the first method is a method for viewing the contrast of an aperture image taken by a camera.
  • a diaphragm 502 is arranged at a position conjugate with the sample on the bottom surface of the glass container 501, and an image of the diaphragm 502 is formed at the position of the sample (bottom surface of the glass container).
  • the camera 503 captures an image of the aperture reflected from the glass surface.
  • a stage scan is performed to move the glass container or objective lens up and down, and the sample image is focused by focusing on the aperture image.
  • the problem with the first method is that a stage scan is required for focusing and it takes time. Depending on the stage scan speed, it takes about tens of seconds. In addition, there is a problem that the range in which the aperture image can be seen is narrow even if the speed is increased by attaching two cameras to a multifocal system and eliminating stage scanning.
  • FIG. 31 is a diagram for explaining a conventional second method.
  • the second method is a method of focusing by looking at the reflection position on the glass surface. Light from the LED 561 passes through the objective lens 523 and enters the sample 522 at an angle. Then, the position of the reflected light is acquired by the CCD camera 564. Since the second method can be controlled so that the position of the beam is at the center, it is possible to always maintain the focus.
  • the problem with the second method is that the accuracy of focusing is poor.
  • the phenomenon of inaccuracy is caused by distortion of the optical system.
  • the position of the incident light is shifted by X / (magnification). Since this deviation occurs due to various external factors such as heat and vibration, it is necessary to adjust the offset every time.
  • the range and accuracy of focusing depend on the angle at which the beam is incident on the glass surface S. For this reason, there is a pinch between the accuracy and the range.
  • JP2015-227940A Japanese Patent No. 5612259
  • An object of the present invention is to provide an autofocus device capable of autofocusing with high accuracy, a wide range, and high speed even in a microscope having a large numerical aperture, and an optical device and a microscope including the autofocus device.
  • the present disclosure relates to an autofocus device used in an optical device that includes a stage that supports a transparent member on which an observation object is placed, and a magnifying optical system that observes the observation object.
  • the autofocus device is a light source device that emits light to an observation object via a magnifying optical system, and a radiated light emitted from the light source device that is disposed at a position opposite to the observation object with respect to the magnifying optical system.
  • Light detection that receives the reflected light reflected by the reflecting surface from the light source device that reaches the reflecting surface of the transparent member via the shielding surface and the enlarging optical system via the enlarging optical system
  • the control device determines the position of the stage or the magnifying optical system based on the reflected light of the shielding object obtained by making the radiation light from the light source device limited by the shielding object incident on the observation object under a plurality of different conditions. Adjust.
  • the light source device is configured to be able to variably adjust the angular distribution of light emitted to the shield.
  • the autofocus device further includes an optical element that reflects part of the emitted light from the light source device limited by the shield.
  • the emitted light from the light source device reflected by the optical element enters the observation object.
  • the autofocus device further includes an optical element that shields, reduces, or reflects a part of the emitted light from the light source device limited by the shield. Radiated light from the light source device that has passed without being shielded, dimmed or reflected by the optical element is incident on the observation object.
  • control device determines a control target value from the position of the reflection image of the shielding object obtained under a plurality of different conditions, and adjusts the position of the stage or the magnifying optical system.
  • control device determines a control target value from the light intensity obtained by dividing and integrating a plurality of shielding object images obtained under a plurality of different conditions, and adjusts the position of the stage or the magnifying optical system.
  • the present disclosure relates to an autofocus device used in an optical device that includes a stage that supports a transparent member on which an observation object is placed and a magnification optical system that observes the observation object.
  • the autofocus device is a light source device that emits light to an observation object via a magnifying optical system, and a radiated light emitted from the light source device that is disposed at a position opposite to the observation object with respect to the magnifying optical system.
  • Shielding object to be limited, and imaging device that receives reflected light reflected by the reflecting surface from the light source device that reaches the reflecting surface of the transparent member via the shielding object and the magnifying optical system via the magnifying optical system
  • a control device for controlling the position of the stage or the magnifying optical system.
  • the light source device emits light at a non-zero angle with respect to the axis of the magnifying optical system.
  • the control device adjusts the position of the stage or the magnifying optical system so that the position of the image of the shield imaged by the imaging device matches the target position.
  • control device adjusts the position of the stage or the magnifying optical system so that the position of the opening of the shielding object in the image of the shielding object photographed by the imaging device matches the target position.
  • control device separates the inside and the outside of the opening in the image of the shielding object by performing image processing on the image obtained by the imaging device.
  • the light source device is configured such that the angle of light emitted with respect to the shield is variable.
  • the control device emits light at a first position that is a position of an image of the shielding object when the light source device emits light at a first angle and at a second angle different from the first angle to the light source device.
  • the position of the stage or the magnifying optical system is adjusted so that the difference from the second position, which is the position of the image of the shielding object in the case of this, becomes the target value.
  • the light source device is configured such that the angle of light emitted with respect to the shield is variable.
  • the control device coarsely adjusts the position of the stage or the magnifying optical system based on the first position that is the position of the image of the shielding object when the light source device emits light at the first angle, and the light source device receives the first light source device.
  • the position of the stage or the magnifying optical system is finely adjusted based on the second position that is the position of the image of the shielding object when light is emitted at a second angle that is larger than this angle.
  • the light source device is configured to be able to change the angle at which the light emitted from the light source device receives light emitted from the light source device and receives the light emitted from the light source device toward the shielding object.
  • an electro-optical element The control device changes the angle of the electric optical element depending on whether the angle of the light emitted from the light source device is the first angle or the second angle.
  • the magnifying optical system includes an objective lens, a half mirror, an optical path through which the half mirror transmits, an optical path through which the half mirror reflects, and an optical path through which the half mirror transmits. And a camera-side imaging lens disposed on the other of the optical paths reflected by the half mirror.
  • the light source device emits polarized light
  • the magnifying optical system is disposed between the objective lens, the quarter-wave plate, the polarization beam splitter, the quarter-wave plate and the polarization beam splitter. Imaging lens.
  • control device adjusts the position of the stage or the magnifying optical system based on the coordinates of the center of gravity of the image of the aperture in the image of the shielding object.
  • control device adjusts the position of the stage or the magnifying optical system based on the coordinates of the edge of the image of the opening in the image of the shielding object.
  • the present disclosure relates to an optical device including a stage, a magnifying optical system, and any one of the autofocus devices described above.
  • the present disclosure relates to a microscope including a stage, a magnifying optical system, and any one of the above autofocus devices.
  • the present invention it is possible to realize autofocus capable of autofocus with high accuracy, wide range, and high speed.
  • imaging with a microscope having a large numerical aperture can be easily automated.
  • FIG. 1 it is a figure for demonstrating the modification at the time of using a slit instead of an aperture_diaphragm
  • FIG. 3 is a flowchart for illustrating autofocus control in the first embodiment.
  • FIG. 10 is a diagram illustrating the principle of autofocus executed in the second embodiment. In Embodiment 2, it is a figure for demonstrating the modification at the time of using a slit instead of an aperture_diaphragm
  • restriction. 10 is a flowchart for illustrating autofocus control in the second embodiment. It is the image of the slit caught with the camera for AF.
  • 6 is a diagram illustrating a configuration of an optical system of a microscope according to a third embodiment. FIG. It is an image in case an incident angle is a positive angle. It is an image in case an incident angle is a negative angle.
  • FIG. 10 is a flowchart for illustrating autofocus control in the third embodiment.
  • FIG. 6 is a diagram illustrating a configuration of an optical system of a microscope according to a fourth embodiment.
  • FIG. 19 is a diagram showing a positional relationship between the shape of the rotary mirror RM1 and the light B in FIG. In the microscope 201 shown in FIG. 18, it is a figure which shows the state which the rotation mirror RM1 rotated 180 degrees.
  • FIG. 21 is a diagram showing a positional relationship between the state of the rotating mirror RM1 shown in FIG.
  • FIG. 10 is a diagram illustrating a configuration of an optical system of a microscope according to a fifth embodiment.
  • FIG. 23 is a diagram showing a state in which a rotary mask RM2 is rotated by 180 ° in the microscope 251 shown in FIG.
  • FIG. 25 is a diagram showing a positional relationship between the state of the rotary mask RM2 shown in FIG.
  • FIG. 10 is a diagram illustrating a configuration of an optical system of a microscope according to a sixth embodiment.
  • FIG. 27 is a diagram showing a positional relationship between the shape of the rotary mask RM3 and the light B in FIG.
  • the microscope 271 shown in FIG. 26 it is a figure which shows the state which the rotation mask RM3 rotated 180 degrees.
  • It is a figure which shows the positional relationship of the state of the rotation mask RM3 shown in FIG.
  • It is a figure for demonstrating the conventional 1st method.
  • It is a figure for demonstrating the conventional 2nd method.
  • FIG. 1 is a diagram illustrating a configuration of an optical system of a microscope according to the present embodiment.
  • Light from the AF laser 11 is reflected by the electric mirror M and enters the iris IR. Since the diaphragm IR and the electric mirror M are in a conjugate relationship, the incident angle ⁇ of light to the diaphragm IR can be controlled by the angle ⁇ of the electric mirror M.
  • the light from the iris IR passes through the imaging lens L2 and the objective lens OL, and is reflected by the surface of the glass 19 (glass surface S) on which the sample 20 is placed.
  • the reflected light is imaged by the autofocus camera (AF camera) 22 through the objective lens OL and the imaging lens L1, and an image of the iris IR is reflected on the AF camera 22.
  • AF camera autofocus camera
  • the feature of this embodiment is that the shaped light is irradiated at an angle with respect to the glass surface S on which the sample 20 is placed on the stage ST, and the position of the reflected image is observed.
  • the sample is irradiated with light vertically without an angle, and the reflection image is blurred.
  • the present embodiment is different in that light is incident at an angle ⁇ and the position of the reflected image is viewed.
  • the incident light is incident at an angle ⁇ , and the position of the image of the iris IR in the reflected image is measured, so that it can be determined in which direction and how far the focal point is from the glass surface S. This leads to faster focusing.
  • Patent Document 2 On the other hand, in the second method shown in Patent Document 2, light is incident without being shaped and the position of the center of gravity of the light is observed.
  • the method of the present embodiment is different in that incident light is shaped using an iris IR. Since the shape of the image is determined by the iris IR, the reflected image derived from the sample can be erased by image processing. This leads to higher accuracy of autofocus.
  • Patent Document 2 also differs in that the straightness of the light applied to the glass surface is low. If the straightness of the light is low, the image changes remarkably due to a change in focus, so it is easy to lose reflected light, leading to a reduction in range.
  • this method is different in that the incident angle can be adjusted. By adjusting the incident angle, the balance between range and accuracy can be adjusted.
  • the autofocus device according to the present embodiment is useful for easily realizing automation of a microscope.
  • a microscope 1 shown in FIG. 1 includes a stage ST, an autofocus optical system, and an observation optical system.
  • the microscope 1 is a light source including an AF laser 11, an electric mirror M, and a Kepler beam expander 13, an aperture IR, an imaging lens L2, a half mirror HM, and a dichroic mirror DM as an autofocus optical system. And an objective lens OL, an imaging lens L1, and an AF camera 22.
  • the microscope 1 further includes an excitation filter 23, an observation dichroic mirror 24, absorption filters 25 and 28, an imaging lens 26, and an observation camera 27 as an observation optical system.
  • the laser light reflected by the electric mirror M passes through the beam expander 13 and enters the aperture IR.
  • the incident angle ⁇ of the laser to the iris IR can be controlled by the angle ⁇ of the electric mirror M. Since the diaphragm IR and the glass surface S are in a conjugate position, a shadow of the diaphragm IR is imaged on the glass surface S. The image of the iris IR is reflected and projected onto the AF camera 22.
  • the observation optical system light having a wavelength that passes through the AF dichroic mirror DM and the absorption filter 28 can be used.
  • the sample 20 can be irradiated with light through the observation dichroic mirror 24. Then, fluorescence or reflected light can be observed with the observation camera 27.
  • the absorption filter 28 absorbs only the light from the AF light source and prevents the leaked light from the AF light source from entering the observation camera 27.
  • the microscope 1 further includes a control device 100 that controls the angle ⁇ of the electric mirror M, the position of the aperture IR in the D direction, and the position of the stage ST.
  • the control device 100 may control not only the position of the stage ST but also the position or both of the objective lens OL and other lenses (lens before the camera and before the aperture). In the following, for the sake of explanation, the control device 100 proceeds with a method of controlling the position of the stage ST.
  • FIG. 2 is a block diagram showing objects to be controlled by the control device.
  • the control device 100 adjusts the position of the aperture IR in the D direction by using the aperture position adjustment unit 101.
  • the control device 100 adjusts the angle ⁇ of the electric mirror M by the mirror angle adjustment unit 102.
  • the control device 100 drives the stage position adjusting unit 103 based on the position in the image of the aperture IR image captured by the AF camera 22 and controls the position of the stage ST in the Z direction.
  • FIG. 3 is a diagram showing the principle of autofocus executed in the first embodiment.
  • the laser light is incident on the sample 20 at an incident angle corresponding to the incident angle ⁇ to the diaphragm IR.
  • the AF camera 22 observes the image of the iris IR as in the camera image P1.
  • the position of the image of the iris IR also shifts left and right in the image.
  • the target position XT of the iris IR image corresponding to the in-focus position is determined in advance, and the difference dx between the position X1 of the iris IR image obtained from the camera image P1 and the target position XT is calculated.
  • the in-focus position is when the difference dx is zero. Since the direction and amount of movement of the stage ST on which the glass surface S is placed can be known from the difference dx, it is possible to always focus at high speed.
  • the accuracy is deteriorated. This is because the reflected image deteriorates the positioning accuracy of the image of the iris IR.
  • the image of the aperture portion of the aperture IR is an image depending on a sample such as a cell, and the reflected image of the aperture IR is not uniform. For this reason, when the position of the image of the iris IR is determined by the weighted center of gravity such as a divided photodiode, the focus is shifted depending on the image of the sample.
  • the image is separated by image processing (for example, binarization processing, contour extraction processing, etc.) so that the aperture is a white image and the shielding portion by the iris IR is a black image. It is preferable to do. This enables highly accurate autofocus independent of the sample.
  • FIG. 4 is a diagram for explaining a modification in the first embodiment in which a slit is used instead of a circular diaphragm.
  • the difference dx is calculated from the position of the weighted center of gravity of the image of the iris IR.
  • the X coordinate of the image is used as it is, or X1 is calculated by a simple calculation.
  • the difference dx can be calculated.
  • the shape of the diaphragm IR may be various shapes such as a star shape and a polygonal shape in addition to the circular shape in FIG. 3 and the slit shape in FIG.
  • FIG. 5 is a diagram showing the relationship between the position of the center of gravity of the aperture opening image and the position of the stage when the incident angle is ⁇ L.
  • FIG. 6 is a diagram showing the relationship between the position of the center of gravity of the aperture opening image and the position of the stage when the incident angle is ⁇ H (> ⁇ L).
  • the horizontal axis indicates the pixel position (px: pixel) indicating the center of gravity of the aperture IR aperture image
  • the vertical axis indicates the position in the Z direction ( ⁇ m) of the stage that moves the glass surface of the sample. ).
  • the incident angle ⁇ ⁇ H (when the incident angle ⁇ is large), the amount of movement of the beam increases and the accuracy increases (about 50 nm / px).
  • the range Z-direction range in which autofocus can be performed
  • the range Z-direction range in which autofocus is possible) becomes wide.
  • FIG. 7 is a schematic diagram of an optical system of the autofocus device according to the present embodiment.
  • FIG. 8 is a flowchart for explaining autofocus control in the first embodiment.
  • the focus position has a linear relationship with the position of the aperture IR in the optical axis direction.
  • the aperture IR is at the focal position of the imaging lens L2
  • the glass surface S is focused. Therefore, the position of the iris IR is uniquely determined as DT from the distance from the glass surface S.
  • Other parameters to be set in advance include electric mirror angles ⁇ L and ⁇ H corresponding to coarse and fine movement stage control, respectively, and convergence determination values ⁇ L and ⁇ H for determining the in-focus corresponding to each. These are basically constant values determined by the developer of the autofocus device regardless of the user.
  • step S1 the control device 100 sets the position D of the iris IR to the position DT.
  • step S2 the control device 100 determines to set the incident angle ⁇ to the iris IR to ⁇ L having a high accuracy.
  • step S4 the control device 100 acquires a reflected image by the AF camera 22, and calculates the position X1 of the center of gravity of the iris IR.
  • step S7 the control device 100 determines whether or not the difference dx is smaller than the determination convergence value ⁇ . If the difference dx ⁇ is not satisfied in step S7 (NO in S7), the processes of steps S3 to S6 are executed again.
  • the incident angle ⁇ is set to a non-zero angle, it is possible to immediately calculate the moving direction and moving amount of the stage from the position of the image of the iris IR. It becomes possible.
  • the accuracy and range of auto-focusing is adjusted by controlling the incident angle ⁇ of light on the sample 20 to ⁇ L and ⁇ H by the electric mirror M. it can.
  • the incident angle ⁇ to the diaphragm IR is large, the gravity center position of the image of the diaphragm IR on the glass surface S greatly moves when the glass surface S is moved in the vertical direction.
  • the image of the iris IR is easily deviated from the AF camera 22 and the range is narrowed, but the accuracy of the image is increased because the position variation of the image is increased.
  • the incident angle ⁇ to the iris IR is small, the accuracy is deteriorated, but the range is widened.
  • the incident angle ⁇ is reduced and auto-focusing is performed over a wide range, and then the incident angle ⁇ is increased and auto-focusing is performed with high accuracy. In this way, autofocus that achieves both a wide range and high accuracy is realized.
  • the outline of the procedure for using the autofocus device is as follows. First, the user decides where to focus from the glass surface S. Based on the position determined by the user, the position of the iris IR in the D direction is moved. Then, the angle ⁇ of the electric mirror M is set so as to give the incident angle ⁇ L, a reflection image of the aperture IR is taken by the AF camera 22, and the position X1 of the center of gravity is calculated. Then, after setting the angle ⁇ of the electric mirror M so as to give the incident angle ⁇ L, a reflected image is taken again, and the position X2 of the center of gravity is calculated.
  • FIG. 9 is a diagram showing the principle of autofocus executed in the second embodiment.
  • the laser beam B1 is incident on the sample 20 at an incident angle corresponding to the incident angle ⁇ to the diaphragm IR.
  • the AF camera 22 observes the image of the iris IR as in the camera image P1.
  • the position of the image of the iris IR also shifts left and right in the image.
  • the laser beam B2 is incident on the sample by changing the angle of the electric mirror M so that the incident angle to the iris IR is - ⁇ opposite to the incident angle ⁇ .
  • the image of the diaphragm IR similarly to the angle ⁇ , when the Z position of the glass surface S from the focal position changes, the image of the diaphragm IR also changes the position in the image.
  • the position of the image of the iris IR shifts in the opposite direction.
  • the in-focus position is when the difference dx in the position of the image of the iris IR at the incident angle ⁇ and the incident angle ⁇ is zero. Since the direction d and the amount of movement of the stage ST on which the glass surface S is placed are known from the difference dx between the positions of the images of the iris IR between the camera image P1 and the camera image P2, it is possible to always focus at high speed. .
  • the accuracy is deteriorated. This is because the reflected image deteriorates the positioning accuracy of the image of the iris IR.
  • an image of a sample such as a cell to be observed is visible at the opening of the iris IR. Therefore, the image of the aperture portion of the aperture IR is an image depending on a sample such as a cell, and the reflected image of the aperture IR is not uniform. For this reason, when the position of the image of the iris IR is determined by the weighted center of gravity such as a divided photodiode, the focus is shifted depending on the image of the sample. To solve this problem and improve autofocus accuracy, the image is separated by image processing (for example, binarization processing, contour extraction processing, etc.) so that the aperture is a white image and the shielding portion by the iris IR is a black image. It is preferable to do. This enables highly accurate autofocus independent of the sample.
  • image processing for example, binarization processing, contour extraction processing, etc.
  • a photographed image at an incident angle ⁇ and an incident angle ⁇ can be used as a method of matching the image of the iris IR with the target position XT.
  • the user does not need to set the target position XT of the image of the iris IR in advance.
  • the accuracy and range can be adjusted by changing the incident angle ⁇ to the iris IR during autofocus. Since the adjustment of the accuracy and the range has been described with reference to FIGS. 4 and 5, the description thereof will not be repeated here.
  • FIG. 10 is a diagram for explaining a modification in the second embodiment in which a slit is used instead of a diaphragm.
  • the difference dx is calculated from the position of the weighted center of gravity of the image of the iris IR.
  • the X coordinate of the image is used as it is, or X1 and X2 are calculated by simple calculation.
  • the difference dx can be calculated.
  • the shape of the iris IR may be various shapes such as a star shape and a polygonal shape in addition to the circular shape and the slit shape.
  • a sample is sequentially photographed by irradiating the sample with laser light from two directions of incident angles ⁇ and ⁇ to the aperture IR at each of an incident angle with a wide range and an incident angle with a narrow range. Then, the center of gravity of the image of the iris IR is obtained.
  • the stage ST position where the aperture IR and the AF camera are conjugate is not known. For this reason, it is necessary to set in advance where the center of gravity of the iris IR is located and whether the iris IR and the AF camera are conjugate.
  • the difference is zero, so it is not necessary to set the reference position in advance.
  • FIG. 11 is a flowchart for explaining autofocus control in the second embodiment.
  • the position DT of the iris IR is determined according to how far the glass surface S is focused, and corresponds to the stage control of coarse movement and fine movement, respectively.
  • the angles .theta.L and .theta.H of the electric mirrors to be determined are determined, and convergence determination values .epsilon.L and .epsilon.H for determining in-focus are determined in advance.
  • step S11 the control device 100 sets the position D of the iris IR to DT.
  • step S12 the control device 100 decides to set the incident angle ⁇ to the iris IR to ⁇ L having a high accuracy.
  • the convergence determination value ⁇ is ⁇ L.
  • step S14 the control device 100 acquires a reflected image by the AF camera 22, and calculates the position X1 of the center of gravity of the iris IR.
  • the control device 100 determines whether or not the difference dx is smaller than the determination convergence value ⁇ . If the difference dx ⁇ is not satisfied in step S19 (NO in S19), the processes in steps S13 to S18 are executed again.
  • the in-focus position of the iris IR can be known without setting the target position XT in advance.
  • the position of the stage ST where the aperture IR and the AF camera are conjugate is not known. Therefore, it is necessary to set in advance in which position on the AF camera the center of gravity of the iris IR is conjugate with the iris IR and the AF camera.
  • the position of the glass surface S conjugate with the aperture IR is a position where the difference becomes zero, so that it is not necessary to set the target position XT in advance.
  • the conventional autofocus method shown in FIG. 31 is performed by making light incident on the glass surface from one direction and detecting a change in the position of the return light.
  • the focus is shifted by “ ⁇ X / magnification”.
  • magnification is 100 times
  • the XY direction of the iris IR direction perpendicular to the axis of the magnifying optical system
  • the focus position is shifted by 1 ⁇ m in the optical axis direction.
  • the focus position is shifted by several hundred nm, the obtained image is greatly blurred. Therefore, in the conventional autofocus, it is necessary to adjust the offset every time, and automation during observation is difficult.
  • photographing is performed from two directions with respect to the diaphragm, and a difference between the two diaphragm positions is detected. Therefore, there is no problem even if the position of the iris IR is shifted in the XY direction.
  • the position of the stop IR is also shifted in the D direction (optical axis direction), but when it is shifted by ⁇ D in the D direction, the focus position becomes “ ⁇ D / magnification 2 ”.
  • the magnification is 100 times
  • the focus is set on the glass surface S.
  • the focus position can be shifted from the glass surface S by changing the position of the aperture IR in the D direction.
  • the distance from the glass surface S to the autofocus focal plane and the position of the iris IR in the D direction are linear. For this reason, it is possible to specify the position to be focused from the glass surface by the distance. It is also possible to control the position of the focal plane without changing the position of the stop.
  • the focus position changes linearly if the difference dx between the center of gravity of the apertures of the two images is controlled to a non-zero value.
  • the position of the aperture IR in the XY direction (direction orthogonal to the axis of the magnifying optical system)
  • the Z position of the stage is changed for focusing.
  • focusing may be performed by moving the objective lens or another lens (lens before the camera or the lens before the diaphragm).
  • the autofocus device of the present embodiment can be applied to a microscope incorporated in an industrial facility in addition to a research microscope.
  • FIG. 12 is an image of the slit captured by the AF camera.
  • This image is an image of one slit SL, but many vertical stripes are generated inside the slit SL due to interference.
  • the reflected image of the slit SL is more blurred as light is scattered from the slit SL at the left edge portion than at the right edge.
  • This phenomenon is the same not only for the slit SL but also for the image of the iris IR. This phenomenon makes it difficult to accurately detect the gravity center positions of the slit SL and the diaphragm IR by image processing.
  • the edge on the non-scattering side is determined in advance depending on whether the incident angle ⁇ is positive or negative. Therefore, in the embodiment, instead of calculating the center of gravity of the image, the edge position on one side of the slit image is detected to obtain information for autofocus.
  • FIG. 13 is a diagram illustrating a configuration of an optical system of the microscope according to the third embodiment.
  • a microscope 105 shown in FIG. 13 includes a stage ST, an autofocus optical system, and an observation optical system.
  • the microscope 105 is an optical system for autofocus, and includes a light source including an AF laser 11 and an electric mirror M, a slit SL, a polarization beam splitter BS, an imaging lens L11, a lens L12, and a quarter-wave plate QR.
  • the microscope 105 further includes an excitation filter 23, an observation dichroic mirror 24, an absorption filter 25, an imaging lens 26, and an observation camera 27 as an observation optical system.
  • information for aligning the position of the stage ST is obtained using the laser light reflected by the electric mirror M.
  • the sample 20 can be irradiated with light from a light source (not shown) through the observation dichroic mirror 24. Then, fluorescence or reflected light can be observed with the observation camera 27.
  • the microscope 105 further includes a control device 110 that controls the angle ⁇ of the electric mirror M, the D-direction position of the slit SL, and the position of the stage ST.
  • the control device 110 may control not only the position of the stage ST but one or a plurality of positions of the objective lens OL, the imaging lens L11, and the lens L12.
  • the light from the AF laser 11 is reflected by the electric mirror M and enters the slit SL.
  • the incident angle ⁇ of light to the slit SL can be controlled by the angle ⁇ of the electric mirror M.
  • the light from the slit SL is reflected by the polarizing beam splitter BS, passes through the imaging lens L11 and the quarter-wave plate QR, is reflected by the dichroic mirror DM, and passes through the objective lens OL, and the glass 19 on which the sample 20 is placed. To the glass surface S. In this way, the slit image is projected onto the glass surface S.
  • the laser light is reflected by the polarization beam splitter BS.
  • the laser light is polarized when it is emitted from the AF laser 11, and almost 100% of the light is reflected by the polarization beam splitter BS by aligning the direction of the AF laser 11.
  • the incident laser light and reflected light can be acquired by the AF camera 22 without loss.
  • the light that reaches the surface of the glass 19 is reflected by the surface of the glass 19.
  • This reflected light is reflected by the dichroic mirror DM after passing through the objective lens OL, passes through the quarter-wave plate QR, the imaging lens L11, the polarizing beam splitter BS, the filter F, and the lens L12, and is an autofocus camera. (AF camera) 22 forms an image. In this way, the AF camera 22 displays an image of the slit SL.
  • the imaging lenses L1 and L2 in FIG. 1 can be combined into one imaging lens L11, and the number of lenses can be reduced by one.
  • the quarter wavelength plate QR is transmitted to the AF camera 22 through the polarization beam splitter BS by the quarter wavelength plate QR. Without the wave plate QR, the return light from the imaging lens L11 enters the AF camera 22 and becomes noise. Further, if the quarter wavelength plate QR is not installed at a certain angle, the reflected light from the quarter wavelength plate QR enters the AF camera 22 and becomes noise.
  • the position of autofocus can be shifted from the glass surface by moving the imaging lens L11 or the lens L12 back and forth.
  • the lens L12 is installed outside the microscope so that the offset can be adjusted from the outside.
  • the configuration for controlling the imaging lens L11 inside the microscope is employed, the lens L12 is not necessary.
  • the dichroic mirror DM can separate the autofocus optical path and the observation optical path. Therefore, autofocus while observing with the microscope 105 is possible.
  • the filter F near the lens L12 is for removing light other than autofocus. Without this, the observation light enters the AF camera 22, which causes a problem in autofocus during observation.
  • the light shaped by the slit SL is irradiated at an angle to the glass surface S on which the sample 20 is placed on the stage ST, and the position of the reflected image is observed.
  • FIG. 12 illustrates that the edge disturbance is noticeable only on one edge, but it was found that the edge where the disturbance is remarkable is on a different side depending on whether the incident angle ⁇ is positive or negative. This is presumably because the light incident on the glass surface is scattered for some reason when reflected, and scattering occurs on the reflected light side. Therefore, the edge on the non-scattering side is determined in advance depending on whether the incident angle ⁇ is positive or negative.
  • FIG. 14 is an image when the incident angle is a positive angle.
  • the incident angle is a positive angle (+ ⁇ )
  • the left side is clearly seen when the edge is observed.
  • FIG. 15 is an image when the incident angle is a negative angle.
  • the incident angle is negative ( ⁇ )
  • the right side can be clearly seen when the edge is observed.
  • FIG. 16 is a diagram for explaining edge detection.
  • the image taken by the AF camera 22 is an image in which the left edge is disturbed
  • the luminance of the pixel is scanned from the right side of the image, and the position exceeding the threshold is set as the edge. If both edges are clearly visible, the center of gravity is sufficient, but one of the edges is disturbed, so this method is adopted.
  • the edge detection method is also useful when using objective lenses that are not submerged or submerged.
  • objective lenses that are not submerged or submerged.
  • light between the solution and the glass surface is reflected.
  • the light on the lower surface and the upper surface of the glass is reflected, and the reflected images overlap.
  • the image is averaged in the Y direction to make it one-dimensional.
  • one-dimensional data is scanned from the left side to search for a location that exceeds a specified threshold value.
  • a scan is performed from the right to search for a location exceeding the threshold (see FIG. 16). If the threshold is not exceeded, it is assumed that autofocus has failed. Since the image processing method is simple, it can be operated at high speed.
  • FIG. 17 is a flowchart for explaining the autofocus control in the third embodiment. Also in the third embodiment, as in the first and second embodiments, the position DT of the imaging lens L11 and the angle ⁇ of the electric mirror are determined according to how far the glass surface S is focused. A convergence determination value ⁇ for determining in-focus is determined in advance. Whether the program has completed autofocus can be known from the convergence judgment value, and automatic shooting is prevented from starting before autofocus.
  • step S101 the control device 110 sets the position D of the imaging lens L11 to DT and sets the incident angle ⁇ to the slit SL. Further, the convergence determination value is ⁇ .
  • step S102 the control device 110 sets the angle ⁇ of the electric mirror M to an angle that becomes the incident angle ⁇ .
  • the control device 110 acquires a reflected image by the AF camera 22, and detects the position EL of the left edge of the slit SL.
  • control device 110 changes the angle ⁇ of the electric mirror M so that the incident angle becomes ⁇ in step S104, acquires a reflected image again in step S105, and calculates the position ER of the right edge of the slit SL.
  • step S108 the control device 110 determines whether or not the difference dx is smaller than the determination convergence value ⁇ . As a result, the user and the main program can know whether or not the convergence value has been reached. Note that the position of the objective lens OL or the like may be adjusted instead of moving the stage ST.
  • control device 110 ends autofocus.
  • the autofocus device according to the third embodiment has the same effects as the autofocus device according to the second embodiment.
  • the autofocus device of the third embodiment can further increase the autofocus accuracy.
  • the sample is irradiated with laser light from two directions of incident angles ⁇ and ⁇ to the slit SL and sequentially photographed, and the edge of the image of the slit SL on the side corresponding to the positive / negative of the incident angle is obtained. Then, the difference between the edge positions is taken, and the vertical position of the glass surface S is controlled so that the difference is close to the actual slit width.
  • this method is used, accurate focusing can be performed even when one edge of the image of the aperture or slit SL is disturbed.
  • the incident angle ⁇ is variable.
  • the incident angle ⁇ is set to a fixed non-zero angle, and only a clear edge is targeted. You may control so that it may correspond with a position.
  • the edge used for focusing is the edge corresponding to the positive or negative of the incident angle ⁇ .
  • Embodiment 4 In Embodiments 1 to 3, the light from the AF light source is incident on the magnifying optical system at a non-zero fixed or variable incident angle ⁇ , so that the position of the aperture or slit focused on the AF camera is focused. The difference in position was used for focusing.
  • FIG. 18 is a diagram illustrating a configuration of an optical system of the microscope according to the fourth embodiment.
  • a microscope 201 shown in FIG. 18 includes a stage, an autofocus optical system, and an observation optical system. The details of the stage and the optical system for observation are the same as those in Embodiments 1 to 3, and are not shown here.
  • FIG. 18 shows only the optical system for autofocus.
  • the microscope 201 is a light source 211, a slit SL or a diaphragm IR, a rotating mirror RM1, an imaging lens L211, a dichroic mirror DM, an objective lens OL, a sensor 222, and a control device as an autofocus optical system. 210.
  • the sensor 222 may use the same AF camera as in the first to third embodiments, but may use a split type light receiving element in which the attendance surface is divided.
  • the control device 210 determines a control target value from the light intensity obtained by dividing and integrating a plurality of shielding object images obtained under a plurality of different conditions, and Adjust the position.
  • information for aligning the position of the stage ST is obtained using light from the light source reflected by the rotating mirror RM1.
  • FIG. 19 is a diagram showing a positional relationship between the shape of the rotary mirror RM1 and the light B in FIG.
  • the rotating mirror RM1 is configured to rotate about the rotation axis RA1.
  • the rotating mirror RM1 includes, for example, patterns RP1 and RP2 such as aluminum formed on a transparent disk glass by vapor deposition.
  • the pattern RP1 is configured to reflect light that has passed through the slit SL or the diaphragm IR in a different pattern
  • the pattern RP2 is configured to reflect in a pattern different from the pattern RP1.
  • the left half and the right half are reflected with respect to the optical axis ⁇ . If light near the center is passed, the range will be longer, but the accuracy will be lower. With this in mind, the reflection pattern is designed.
  • the light source 211 may not be a light source with high straightness like a laser.
  • an LED or a mercury lamp may be used. Therefore, the light from the light source 211 may enter the diaphragm IR or the slit SL from all directions.
  • the light B that has passed through the diaphragm IR hits the pattern RP1 of the rotating mirror RM1, and only half of the light B travels to the imaging lens L211.
  • the light transmitted through the imaging lens L211 is reflected by the dichroic mirror DM, passes through the objective lens OL, and is reflected by the glass surface.
  • the reflected light passes through the objective lens OL and the dichroic mirror DM, passes through the imaging lens, and reaches the sensor 222.
  • FIG. 20 is a diagram illustrating a state in which the rotating mirror RM1 is rotated 180 ° in the microscope 201 illustrated in FIG.
  • FIG. 21 is a diagram showing a positional relationship between the state of the rotary mirror RM1 shown in FIG.
  • the rotating mirror RM1 is rotated to alternately change the state of the microscope 201 to the state shown in FIG. 18 and the state shown in FIG. 20, and two images are acquired.
  • control device 210 causes light to enter two different positions across the optical axis of the objective lens OL, and obtains an image corresponding to each light.
  • autofocus with improved accuracy can be realized easily. This is because there is no problem even if the position of the aperture IR or the slit SL on the sensor is shifted due to distortion of the optical system.
  • Embodiment 5 In Embodiment 4, a part of the light after passing through the diaphragm or slit is reflected by the mirror and sent to the objective lens. However, similar focusing can be achieved by blocking a part with a mask instead of reflecting a part with a mirror.
  • FIG. 22 is a diagram illustrating a configuration of an optical system of the microscope according to the fifth embodiment.
  • a microscope 251 illustrated in FIG. 22 includes a stage, an autofocus optical system, and an observation optical system. Since the details of the stage and the optical system for observation are the same as those in Embodiments 1 to 3, illustration is omitted here, and FIG. 22 shows only the optical system for autofocus.
  • the microscope 251 is a light source 211, a slit SL or a diaphragm IR, a rotation mask RM2, a half mirror HM2, an imaging lens L211, a dichroic mirror DM, an objective lens OL, and a sensor as an autofocus optical system. 222 and the control device 210.
  • the senor 222 may use either an AF camera or a divided light receiving element.
  • information for aligning the position of the stage ST is obtained using light from the light source that has passed through the rotary mask RM2.
  • FIG. 23 is a diagram showing a positional relationship between the shape of the rotary mask RM2 and the light B in FIG.
  • rotation mask RM2 is configured to rotate about rotation axis RA2.
  • the rotary mask RM2 includes, for example, patterns RP1 and RP2 such as aluminum formed on a transparent disk glass by vapor deposition.
  • the patterns RP1 and RP2 are configured such that light that has passed through the slit SL or the diaphragm IR has different patterns. In FIG. 23, it is comprised so that half may be interrupted
  • the patterns RP1 and RP2 may reflect light toward the light source like a mirror as long as the light can be blocked.
  • the light source 211 does not have to be high in straightness like the laser as in the fourth embodiment. Therefore, the light from the light source 211 may enter the diaphragm IR or the slit SL from all directions.
  • the light B that has passed through the aperture IR hits the pattern RP2 of the rotary mask RM2, only half of the light B is blocked, and the other half is reflected by the half mirror HM2, and proceeds to the imaging lens L211.
  • the light transmitted through the imaging lens L211 is reflected by the dichroic mirror DM, passes through the objective lens OL, and is reflected by the glass surface.
  • the reflected light passes through the objective lens OL and the dichroic mirror DM, passes through the imaging lens L211 and reaches the sensor 222.
  • FIG. 24 is a diagram showing a state in which the rotary mask RM2 is rotated by 180 ° in the microscope 251 shown in FIG.
  • FIG. 25 is a diagram showing a positional relationship between the state of the rotary mask RM2 shown in FIG.
  • the rotary mask RM2 is rotated to alternately change the state of the microscope 251 to the state shown in FIG. 22 and the state shown in FIG. 24, and two images are acquired.
  • a similar image may be obtained by using, as a mask, an element that electrically controls light transmission, such as liquid crystal, instead of the rotating mask.
  • the mask pattern is a plurality of patterns in which the transmitted light is asymmetric with respect to the optical axis of the magnifying optical system, and the objective lens is irradiated with light and the reflected light is observed.
  • the microscope shown in the sixth embodiment is quite similar to the microscope of the fifth embodiment in that a mask is used, but two imaging lenses are required in a form in which a half mirror is inserted before the imaging lens.
  • FIG. 26 is a diagram illustrating a configuration of an optical system of the microscope according to the sixth embodiment.
  • a microscope 271 shown in FIG. 26 includes a stage, an autofocus optical system, and an observation optical system. The details of the stage and the optical system for observation are the same as in Embodiments 1 to 3, and are not shown here.
  • FIG. 26 shows only the optical system for autofocus.
  • the microscope 271 includes, as an autofocus optical system, a light source 211, a slit SL or a diaphragm IR, imaging lenses L211A and L211B, a rotation mask RM3, a half mirror HM3, a dichroic mirror DM, and an objective lens OL. , Sensor 222 and control device 210.
  • the senor 222 may use either an AF camera or a divided light receiving element.
  • information for aligning the position of the stage ST is obtained using light from the light source that has passed through the rotary mask RM3.
  • FIG. 27 is a diagram showing a positional relationship between the shape of the rotary mask RM3 and the light B in FIG.
  • rotation mask RM3 is configured to rotate about rotation axis RA3.
  • the rotary mask RM3 includes, for example, patterns RP1 and RP2 such as aluminum formed on a transparent disk glass by vapor deposition.
  • the pattern RP1 is configured with a pattern different from the axis of light that has passed through the slit SL or the diaphragm IR
  • the pattern RP2 is configured with a pattern different from the pattern RP1.
  • the patterns RP1 and RP2 may reflect light toward the light source like a mirror as long as the light can be blocked.
  • the light source 211 may not be a laser as long as it emits light of a certain degree of straightness such as an LED. Therefore, the light from the light source 211 may enter the diaphragm IR or the slit SL from all directions.
  • the light B that has passed through the diaphragm IR proceeds to the imaging lens L211.
  • the light transmitted through the imaging lens L211 hits the pattern RP2 of the rotary mask RM3 as shown in FIG. 26, and only half of the light B is blocked and the remaining half is reflected by the half mirror HM3.
  • the light reflected by the half mirror HM3 is reflected by the dichroic mirror DM, passes through the objective lens OL, and is reflected by the glass surface.
  • the reflected light passes through the objective lens OL and the dichroic mirror DM, passes through the imaging lens L211A, and reaches the sensor 222.
  • FIG. 28 is a diagram showing a state in which the rotary mask RM3 is rotated by 180 ° in the microscope 271 shown in FIG.
  • FIG. 29 is a diagram showing the positional relationship between the state of the rotary mask RM3 shown in FIG.
  • the rotary mask RM3 is rotated to alternately change the state of the microscope 271 to the state shown in FIG. 26 and the state shown in FIG. 28, and two images are acquired.
  • a similar image may be obtained by using, as a mask, an element that electrically controls light transmission, such as liquid crystal, instead of the rotating mask.
  • the mask pattern is a plurality of patterns in which the transmitted light is asymmetric with respect to the optical axis of the magnifying optical system, and the objective lens is irradiated with light and the reflected light is observed.
  • the present disclosure provides a stage (ST) that supports a transparent member (19) on which an observation object (20) is placed, and a magnification optical system (L1, L2, HM, DM, OL) that observes the observation object.
  • the present invention relates to an autofocus device used for an optical device having The autofocus device is disposed at a position opposite to the observation object with respect to the light source device (11, M, 13, 211) that emits light to the observation object via the magnification optical system, and the magnification optical system,
  • the shielding light (IR, SL) that restricts the radiated light emitted from the light source device, and the radiated light from the light source device that reaches the reflecting surface of the transparent member via the shielding material and the magnifying optical system is reflected by the reflecting surface.
  • a light detection device (22, 222) that receives the reflected light via the magnifying optical system and a control device (100, 110, 210) that controls the position of the stage or the magnifying optical system.
  • the control device emits the radiated light from the light source device limited by the shielding object under a plurality of different conditions.
  • the position of the stage or the magnifying optical system is adjusted based on the reflected light of the shielding object obtained by the incidence.
  • the light source device is configured to be able to variably adjust the angular distribution of the light emitted to the shield.
  • the autofocus device further includes an optical element (RM1) that reflects a part of the emitted light from the light source device limited by the shield.
  • RM1 optical element that reflects a part of the emitted light from the light source device limited by the shield.
  • the emitted light from the light source device reflected by the optical element enters the observation object.
  • the autofocus device further includes optical elements (RM2, RM3) that shield, reduce or reflect a part of the emitted light from the light source device limited by the shield.
  • optical elements RM2, RM3 that shield, reduce or reflect a part of the emitted light from the light source device limited by the shield.
  • control device (210) determines a control target value from the position of the reflected image of the shielding object obtained under a plurality of different conditions, and adjusts the position of the stage or the magnifying optical system.
  • control device (210) determines a control target value from the light intensity obtained by dividing and integrating the image of the shielding object obtained under a plurality of different conditions, and adjusts the position of the stage or the magnifying optical system.
  • the autofocus device includes a stage (ST) that supports the transparent member (19) on which the observation object (20) is placed, and a magnification optical system (L1, L2, HM) that observes the observation object. , DM, OL).
  • the autofocus device is disposed at a position opposite to the observation target with respect to the magnification optical system and a light source device (11, M, 13) that emits light to the observation target via the magnification optical system.
  • Shielding object (IR) that restricts the radiated light emitted from the reflector, and the reflected light reflected by the reflecting surface through the shielding object and the reflecting surface of the transparent member via the magnifying optical system and the magnifying optical system
  • An imaging device (22) received via the control device and a control device (100) for controlling the position of the stage or the magnifying optical system are provided.
  • the light source device emits light at a non-zero angle ( ⁇ ) with respect to the axis of the magnifying optical system.
  • the control device (100) sets the position of the stage or the magnifying optical system so that the position X1 of the image of the shield imaged by the imaging device (22) matches the target position XT. adjust.
  • control device (100) causes the position X1 or X2 of the opening of the shielding object in the image of the shielding object photographed by the imaging device (22) to coincide with the target position XT. Adjust the position of the stage or magnifying optical system.
  • control device (100) separates the inside and the outside of the opening in the image of the shielding object (IR) by performing image processing such as binarization processing on the image obtained by the imaging device (22). .
  • image processing such as binarization processing
  • the light source device is configured such that the angle ( ⁇ ) of light emitted with respect to the shielding object (IR) is variable.
  • the control device (100) includes a first position (X1) that is the position of the image of the shielding object when the light source device emits light at the first angle ( ⁇ ), and a first angle ( ⁇ ).
  • the difference (dx) from the second position (X1) that is the position of the image of the shielding object (IR) when light is emitted at a second angle ( ⁇ ) different from) is a target value (for example, zero) Adjust the position of the stage or the magnifying optical system so that In this way, it is possible to determine in-focus even if the target position on the captured image corresponding to the in-focus is not set in advance.
  • the light source device is configured such that the angle of light emitted with respect to the shield (IR) is variable.
  • the control device (100) determines the position of the stage or the magnifying optical system based on the first position that is the position of the image of the shielding object (IR) when the light source device emits light at the first angle ( ⁇ L). Coarse adjustment is made to the second position, which is the position of the image of the shield (IR) when the light source device emits light at a second angle ( ⁇ H) that is larger than the first angle ( ⁇ L). Based on this, the position of the stage or the magnifying optical system is finely adjusted. In this way, a wide range and high-precision autofocus can be realized.
  • the light source device can change the angle at which the light emitted from the light source device receives the light emitted from the light source device 11 and the light source 11 that emits light with high straightness and enters the shielding object.
  • the electro-optic element (M) configured in the above.
  • the control device (100) is a motor-driven optical element in which the angle ( ⁇ ) of the light emitted from the light source device is the first angle ( ⁇ or ⁇ L) and the second angle ( ⁇ or ⁇ H). The angle ( ⁇ ) of (M) is changed.
  • the magnifying optical system is a light source disposed in any one of an objective lens (OL), a half mirror (HM), an optical path transmitted through the half mirror (HM), and an optical path reflected by the half mirror (HM).
  • a side imaging lens (L2) and a camera side imaging lens (L1) disposed on the other of the optical path through which the half mirror (HM) transmits and the optical path through which the half mirror (HM) reflects.
  • the light source device emits polarized light
  • the magnifying optical system includes an objective lens (OL), a quarter wave plate (QR), a polarization beam splitter (BS), and a quarter wave plate ( QR) and an imaging lens (L11) disposed between the polarization beam splitter (BS).
  • OL objective lens
  • QR quarter wave plate
  • BS polarization beam splitter
  • QR quarter wave plate
  • L11 imaging lens
  • control device (100) adjusts the position of the stage (ST) or the magnifying optical system based on the coordinates of the center of gravity of the image of the opening in the image of the shielding object.
  • control device (110) adjusts the position of the stage (ST) or the magnifying optical system based on the coordinates of the edge of the image of the opening in the image of the shielding object. If the edge is used, it may be detected with higher accuracy than calculating the center of gravity.
  • the present disclosure relates to an optical device including a stage, a magnifying optical system, and any one of the autofocus devices described above.
  • the present disclosure relates to an optical device including a stage, a magnifying optical system, and any one of the above autofocus devices.

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PCT/JP2019/002216 2018-02-14 2019-01-24 オートフォーカス装置ならびにそれを備える光学装置および顕微鏡 Ceased WO2019159627A1 (ja)

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JP7036396B1 (ja) 2021-12-30 2022-03-15 株式会社 Zido オートフォーカス装置
TWI811758B (zh) * 2020-08-07 2023-08-11 美商奈米創尼克影像公司 用於自動對焦顯微鏡系統之深度學習模組、自動對焦一顯微鏡系統之方法及非暫時性電腦可讀媒體
WO2024181542A1 (ja) 2023-03-01 2024-09-06 国立大学法人大阪大学 受容体型チロシンキナーゼの活性評価方法
WO2025224938A1 (ja) * 2024-04-25 2025-10-30 株式会社Zido オートフォーカス装置

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