WO2018070049A1 - Microscope apparatus, image processing method, and processing device - Google Patents

Abstract

[Problem] To enable operation of an optical element with high accuracy. [Solution] This microscope device includes: an illumination optical system which irradiates a specimen with light; an image forming optical system which forms an image of the light from the specimen; an image capture unit which captures an image; a processing device which processes the image outputted from the image capture unit; and a correction unit which corrects, on the basis of information about the image, the position of a component that constitutes either of the optical systems.

Description

Microscope device, image processing method, and processing device

The present invention relates to a microscope apparatus, an image processing method, and a processing apparatus.

As a microscope apparatus for observing a sample, for example, an apparatus including an illumination optical system that irradiates a sample with light and an imaging optical system that forms an image of light from the sample is known (see, for example, Patent Document 1). ).

Japanese Patent No. 3497229

According to the first aspect of the present invention, an illumination optical system that irradiates light onto a sample, an imaging optical system that forms an image of light from the sample, an imaging unit that captures an image, and an image output from the imaging unit There is provided a microscope apparatus that includes a processing apparatus that processes the above and a correction unit that corrects the position of a component that constitutes one of the optical systems based on image information.

According to the second aspect of the present invention, an illumination optical system that irradiates the sample with light, an imaging optical system that forms an image of light from the sample, an imaging unit that captures an image of the sample by the imaging optical system, A method of processing an image captured by an imaging unit in a microscope apparatus comprising: extracting an image of a component constituting an illumination optical system or an imaging optical system from the image; and extracting the extracted component image And calculating the difference between the position of the component and the target position of the component.

According to the third aspect of the present invention, an illumination optical system that irradiates the sample with light, an imaging optical system that images the light from the sample, an imaging unit that captures an image of the sample by the imaging optical system, A processing apparatus for a microscope apparatus comprising: an image processing unit that extracts an image of a component constituting an illumination optical system or an imaging optical system from an image captured by the imaging unit; and a component extracted by the image processing unit A processing device is provided that includes a calculation unit that calculates a difference between the position of the image of the image and the target position of the component.

It is a perspective view which shows an example of the microscope apparatus which concerns on embodiment. It is a perspective view which shows the detail of the microscope main body shown in FIG. It is a figure which shows the optical path of a microscope main body. It is a perspective view which shows an example of a lens-barrel base, (A) is a figure which shows an external appearance, (B) is a figure which shows an internal structure. It is a figure which shows an example of a shutter operation part, (A) is a perspective view, (B) is an arrow line view in (A). It is a figure which shows an example of a shutter operation part, (A) is a perspective view, (B) is an arrow line view in (A). It is the figure which looked at an example of the internal structure of the base from the bottom face side. It is the figure which looked at an example of the internal configuration of the base from the side. The figure which looked at an example of the internal structure of a base from diagonally downward is shown, (A) is at the time of insertion of a pupil observation optical element, (B) is at the time of retraction | saving of a pupil observation optical element. It is a flowchart which shows an example of the image processing method which concerns on embodiment. (A)-(D) are figures which show an example of the display content of a display apparatus in the case of adjusting the position of a phase ring. (A)-(C) are figures which show an example of the display content of a display apparatus in the case of adjusting the rotation position of a polarizer and an analyzer. (A) And (B) is a figure which shows an example of the display content of a display apparatus in case a bubble is extracted. (A) And (B) is a figure which shows an example of the display content of a display apparatus in the case of making the center of a field stop image and the center of an optical axis correspond.

In the microscope apparatus described in Patent Document 1 described above, many optical elements are arranged on the pupil conjugate plane of the objective lens, and a positioning operation (so-called pupil) for accurately aligning these optical elements with respect to the pupil position. Centering operation) is required. Patent Document 1 described above has a guide function (assist function) that presents an operation of an optical element such as a pupil centering operation to the user. However, simply presenting the operation of the optical element results in the operation of the optical element depending on the user's senses, and individual differences are likely to occur, and it is difficult to accurately adjust (align) the optical element. There is.

Embodiments described below are intended to provide a microscope apparatus, an image processing method, and a processing apparatus that allow a user or the like to accurately operate an optical element such as a pupil centering operation.

Hereinafter, embodiments will be described with reference to the drawings. However, the present invention is not limited to this embodiment. Further, in the drawings, the scale is appropriately changed and expressed, for example, a part thereof is enlarged or emphasized. In each drawing, directions are indicated using an XYZ coordinate system as appropriate. In each of the X direction, the Y direction, and the Z direction, the direction of the arrow is described as the + direction (eg, + X direction), and the opposite direction is described as the − direction (eg, −X direction).

FIG. 1 is a perspective view showing an example of a microscope apparatus 100 according to the present embodiment. FIG. 2 is a perspective view showing details of the microscope main body 10 constituting the microscope apparatus 100. As shown in FIGS. 1 and 2, the microscope apparatus 100 is an inverted microscope, for example. The microscope apparatus 100 includes a microscope main body 10, a first illumination optical system 20, a second illumination optical system 30, an imaging optical system 40, an imaging unit 60, and a processing device 80. FIG. 3 is a diagram illustrating an optical path of the microscope main body 10. In the present specification, the illumination optical system is used in the meaning of both a case where a light source is not included and a case where a light source is included.

The microscope main body 10 includes a base 11, a support column 12, a revolver support unit 13, a driving device 14, and a stage 15. For example, the base 11 is placed on a placement surface such as a table. The base 11 houses optical elements (not shown) such as a reflecting mirror, a relay lens, a filter, and a prism, and forms an optical path over the lens barrel 16 and the lens barrel base 16a. These optical elements form part of the imaging optical system 40 together with the optical elements in the lens barrel 16 and the lens barrel base 16 a, the eyepiece lens 17, and the objective lens 36.

The lens barrel 16 is detachably provided on the base 11 via a lens barrel base 16a. The lens barrel 16 and the lens barrel base 16a and the lens barrel base 16a and the base 11 are fixed by a fastening member (not shown) such as a fixing screw. The lens barrel 16 houses various optical elements such as a plurality of lenses, filters, and prisms. These optical elements form part of the imaging optical system as described above. The lens barrel base 16a has an imaging unit 60 described later. The imaging unit 60 is a so-called camera unit. The lens barrel base 16a has a light splitting mirror or the like disposed therein, branches a part of the optical path of the imaging optical system 40, connects one to the lens barrel 16, and connects the other to the imaging unit 60. ing.

The eyepiece 17 is attached to the upper part of the lens barrel 16. The eyepiece 17 can be replaced with the lens barrel 16. The configuration of the eyepiece 17 is arbitrary. The eyepiece 17 is used for a user to confirm an image visually. Although not shown, an image may be acquired by an imaging element (image sensor) such as a CMOS (Complementary Metal Oxide Semiconductor) or a CCD (Charge Coupled Device) instead of the eyepiece 17. Further, in the base 11, a part of the optical path of the imaging optical system 40 may be branched so that one is connected to the eyepiece 17 and the other is connected to the above-described imaging device. The image acquired by the image sensor may be displayed on a display device such as a liquid crystal display device, or may be captured in a processing device such as a personal computer for image processing.

The operation unit 18 is provided on the base 11. The operation unit 18 is disposed on the side surface of the base 11 so that it can be operated by hand while the user is looking into the eyepiece 17, for example. The operation unit 18 is rotatably provided, and can set the drive amount of the drive device 14 according to the rotation position (or the rotation amount), and can drive and stop the drive device 14. Thereby, the height (position in the Z direction) of the revolver support portion 13 can be adjusted by operating the operation portion 18. The operation unit 18 is not limited to being rotatable. For example, a button for driving the driving device 14 to move the revolver support portion 13 upward or downward may be provided.

As shown in FIG. 2, the connecting portion 19c is a terminal formed on the upper surface of the base 11 and capable of electrical connection. The connection portion 19 c is disposed at a connection portion with the support column 12. The connecting portion 19c is electrically connected to the substrate M arranged in the base 11 via the lead wire 19a. The substrate M is electrically connected to the operation unit 18 via the lead wire 19b. The substrate M includes, for example, a CPU (Central Processing UNIT) or a memory, and constitutes a control unit for each member including the driving device 14. The substrate M receives information operated by the operation unit 18 through the lead wire 19b, and connects a signal for controlling the driving device 14 and power for driving the driving device 14 through the lead wire 19a. To the unit 19c.

The support column 12 is attached to the upper surface of the base 11. The support column 12 is arranged to extend upward (+ Z direction) from the base 11. The support column 12 and the base 11 are fixed by a fastening member (not shown) such as a fixing screw. For example, when mounting a spacer between the two, the support 12 is removed from the base 11 by removing the fastening member. Can be removed. For example, a third illumination optical system is attached to the spacer mounted between the support column 12 and the base 11. Moreover, the support | pillar 12 has the connection part 12c on the lower surface. When the support column 12 is mounted on the base 11, the connection portion 12 c comes into contact with the connection portion 19 c of the base 11 and is electrically connected to the connection portion 19 c.

The connecting portion 12c is electrically connected to a drive source of the drive device 14 via a lead wire (not shown). Further, the connecting portion 12c may be electrically connected to a first illumination optical system 20 or a stage 15 to be described later, and may transmit a signal for controlling these or supply power for driving. The connecting portions 19c and 12c described above can adopt any shape that enables electrical connection when the support column 12 is mounted. For example, the electrical connection may be ensured by contact between the flat portions when the support column 12 is mounted. For example, when one of the pins 12 has a pin shape and the other has a hole shape, Electrical connection may be ensured by being inserted into the hole.

The revolver support unit 13 and the driving device 14 are attached to the side surface on the + X side of the support column 12 and are arranged to extend from the support column 12 in the + X direction. The side surface on the + X side of the support column 12 is a surface facing the direction in which the eyepiece 17 is installed. Therefore, the revolver support unit 13 and the driving device 14 are disposed in a space sandwiched between the support column 12 and the eyepiece lens 17 (lens barrel 16). The revolver support unit 13 holds, for example, a plurality of objective lenses 36 (see FIG. 3) and supports a rotatable revolver.

Rotating the revolver manually or by a drive source such as an electric motor can place any of the plurality of objective lenses in the optical path L (see FIG. 3). In addition, the electric motor etc. which rotate a revolver may be arrange | positioned at the revolver support part 13. FIG. The drive device 14 is an actuator that moves the revolver support unit 13 up and down. The drive device 14 includes, for example, a rotational drive source such as an electric motor, a gear train that transmits the rotation of the electric motor, a cam that is rotated by the gear train, and a link rod that is moved by the cam. The revolver support 13 is moved up and down by the movement of the rod. The drive of the drive device 14 is operated by the operation unit 18 as described above.

However, the configuration of the drive device 14 is arbitrary, and any configuration such as a configuration using a rack and pinion gear together with a drive source or a configuration using a ball screw mechanism is applicable. It should be noted that the revolver support unit 13 may be moved up and down manually by the user instead of using the drive device 14.

The stage 15 is attached to the side surface on the + X side of the support column 12 above the revolver support portion 13 and the driving device 14. The stage 15 holds a sample. The stage 15 includes a through hole penetrating in the vertical direction (Z direction). The sample is held on the stage 15 in a state where it is placed on a transparent plate material such as a glass plate or in a state where it is accommodated in a transparent container such as a glass container. The glass plate or glass container is held on the stage 15 so that the sample is placed in the above-described through hole. The stage 15 may include a jig for holding a glass plate or a glass container in a predetermined position.

Further, as shown in FIG. 2, a support portion 12a fixed to the lens barrel base 16a is disposed on the −X side of the lens barrel 16, and supports the supported portion 15a provided on the + X side of the stage 15. is doing. As described above, the supported portion 15a is supported by the support portion 12a, thereby suppressing the shaking or vibration of the stage 15. The support part 12a and the supported part 15a are fixed by a fastening member (not shown) such as a bolt. For example, when a spacer is mounted between the support part 12a and the supported part 15a, the support part 12a is supported by removing the fastening member. The part 15a can be removed.

Further, the stage 15 may be movable in the X direction and the Y direction (horizontal direction). Further, the stage 15 may be movable in a direction along the vertical direction (Z direction). Such movement of the stage 15 may be performed manually by the user or using a drive source such as an electric motor. When the drive source is used, the control signal related to the movement amount and the power supply to the drive source may be performed from the base 11 via the connection portion 12 c of the support column 12. In addition, an operation unit for operating the movement of the stage 15 may be provided on the base 11.

The first illumination optical system 20 is attached to the upper part of the column 12. The exit side of the first illumination optical system 20 is disposed above the stage 15. As shown in FIG. 3, the first illumination optical system 20 performs transmission illumination on the sample S held on the stage 15. The first illumination optical system 20 includes a light source device 21 and an optical system 22.

The light source device 21 is a light source typified by a halogen lamp that emits visible light (light having a broad wavelength), a white LED, or an LD (laser diode). The light source device 21 may be a light source that emits excitation light (including activation light) that excites a fluorescent substance contained in a sample. The light source device 21 may have one light source or a plurality of light sources. For example, the light source device 21 may include a plurality of light sources having different wavelengths of emitted light, and the wavelength of the light emitted from the light source device 21 may be switchable. Further, the microscope apparatus 100 may not include the light source device 21. For example, the light source device 21 may be provided in the microscope apparatus 100 so as to be replaceable, and may be attached to the microscope apparatus 100 when performing observation.

The optical system 22 is disposed on the light emission side of the light source device 21. As shown in FIG. 3, the optical system 22 includes, for example, a lens 23, a mirror 24, a lens 25, a phase ring 26, a condenser lens 27, and a polarizer (first polarizing plate) 28. Among these, the lens 23, the mirror 24, and the lens 25 are accommodated in, for example, a housing (irradiation head) supported on the upper portion of the column 12. The phase ring 26, the condenser lens 27, and the polarizer 28 are supported by the support column 12 separately from the above-described housing, for example. Further, the lens 23 and the lens 25 may be one optical element, or a plurality of optical elements may be combined. Further, the lens 23 or the lens 25 may be omitted.

The optical system 22 is used for observation using transmission illumination on the sample S. The illumination light emitted from the light source device 21 passes through the lens 23, is reflected by the mirror 24, and then passes through the lens 25. The phase ring 26 is disposed so that it can be inserted into and retracted from the optical path between the lens 25 and the condenser lens 27. The insertion and retraction of the phase ring 26 may be performed manually by the user or by a driving mechanism (not shown). In the phase ring 26, for example, a ¼ wavelength plate that shifts the phase of light from the light source device 21 by ¼λ is formed in a ring shape, and ND that absorbs light inside and outside the ¼ wavelength plate. A filter is placed.

The condenser lens 27 is disposed at a position where light transmitted through the phase ring 26 is incident, and irradiates the sample S with light. For example, the polarizer 28 irradiates the sample S with polarized illumination light emitted from the condenser lens 27 in one direction. The polarizer 28 is disposed so as to be able to be inserted into and retracted from the optical path L. The insertion and retraction of the polarizer 28 may be performed manually by the user or may be performed by a driving mechanism (not shown).

The optical system 22 is not limited to the configuration shown in FIG. 3, and can be changed as appropriate. For example, in FIG. 3, the optical system 22 is an optical system whose optical path is bent by a mirror 24, but may be an optical system that does not include the mirror 24 and has a linear optical path. For example, the optical system 22 may include an optical member such as a diaphragm member. In addition, at least a part of the members included in the optical system 22 may not be included in the optical system 22, and may be included in the light source device 21, for example.

The second illumination optical system 30 is attached to the side surface of the support column 12 on the −X side. The exit side of the second illumination optical system 30 is disposed below the stage 15. The support column 12 is provided with a through hole through which light from the second illumination optical system 30 passes. The second illumination optical system 30 performs epi-illumination on the sample S held on the stage 15 using white light or the like. In addition to performing epi-illumination, the second illumination optical system 30 irradiates the fluorescent material with activation light or excitation light when the microscope apparatus 100 is used as a fluorescent microscope. The second illumination optical system 30 can be exchanged for either epi-illumination or fluorescence observation.

The second illumination optical system 30 includes a light source device 31 and an optical system 32 as shown in FIG. The light source device 31 includes a light source that emits light (illumination light). When the second illumination optical system 30 performs epi-illumination, this light source may be a solid light source such as an LED (light emitting diode) or LD (laser diode), or a lamp light source. When the microscope apparatus 100 is used as a fluorescence microscope, the light source apparatus 31 emits light including activation light that activates the fluorescent substance included in the sample S or light including excitation light that excites the fluorescent substance. Note that the light source device 31 may include one or more light sources. For example, the light source device 31 may include a plurality of light sources having different wavelengths of emitted light, and may be capable of switching the wavelength of light emitted from the light source device 31 according to the type of fluorescent material, for example. Further, the microscope apparatus 100 may not include the light source device 31. For example, the light source device 31 may be provided in the microscope apparatus 100 so as to be replaceable, and may be attached to the microscope apparatus 100 when performing observation.

As shown in FIG. 3, the optical system 32 includes a lens 33, a lens 34, a filter unit 35, and an objective lens 36 on the light emission side of the light source device 31. The optical system 32 is used when illuminating the material S with epi-illumination. (The illumination light (activation light or excitation light) emitted from the light source device 31 is transmitted through the lens 33 and the lens 34). The light enters the filter unit 35.

The filter unit 35 includes a filter 35a, a dichroic mirror 35b, and a filter 35c. The filter 35a has a characteristic of transmitting light having a wavelength such as excitation light incident from the lens 34 and shielding light having a wavelength other than the wavelength such as excitation light. The dichroic mirror 35b has a characteristic of reflecting excitation light and the like incident from the filter 35a and transmitting light (observation light or fluorescence) from an objective lens 36 described later. The light reflected by the dichroic mirror 35 b enters the objective lens 36. The objective lens 36 is disposed at a position where the light reflected by the dichroic mirror 35b is incident, and irradiates the sample S with light. The filter 35c will be described in the imaging optical system 40 described later.

It should be noted that the optical system 32 is not limited to the configuration shown in FIG. For example, in FIG. 3, the optical system 32 may include an optical member such as a diaphragm member. Further, at least a part of the members included in the optical system 32 may not be included in the optical system 32, and may be included in the light source device 31, for example.

The imaging optical system 40 includes an imaging optical system 40a, an imaging optical system 40b, and an imaging optical system 40c. The imaging optical system 40 a guides part of the light (observation light) from the sample S to the image sensor 61. The imaging optical system 40a includes an analyzer (second polarizing plate) 37, an objective lens 36, a filter unit 35, a lens (second objective lens) 41, a mirror 42, a lens 44, a mirror 45, a lens 46, a beam splitter 47, and A lens 48 is provided. These imaging optical systems 40a are accommodated in the base 11 (refer FIG. 2), for example. The image sensor 61 may be housed in the base 11 or may be attached as a camera unit to the side surface of the base 11.

In the present embodiment, the objective lens 36 and the filter unit 35 are members constituting the optical system 32 described above. The analyzer 37 is disposed so as to be able to be inserted into and retracted from the optical path L. The insertion and retraction of the analyzer 37 may be performed manually by the user or may be performed by a driving mechanism (not shown). The analyzer 37 is arranged with the polarization direction orthogonal to the polarizer 28 (crossed Nicols). When the analyzer 37 is inserted into the optical path L, the illumination light emitted from the polarizer 28 is blocked by the analyzer 37 unless the deflection direction is rotated by 90 degrees. The polarizer 28 and the analyzer 37 are not used alone, but both are used to be inserted into or retracted from the optical path L at the same time.

The objective lens 36 is disposed, for example, on a surface including the front focal point (front focal plane) including the rear focal point of the condenser lens 27 (rear focal plane) or in the vicinity thereof. The light (observation light) that has passed through the objective lens 36 passes through the dichroic mirror 35b and enters the filter 35c. The filter 35c has a characteristic of transmitting fluorescence emitted from the sample S and passing through the objective lens 36. The light transmitted through the filter 35 c is reflected by the mirror 42 through the lens 25 and then enters the lens 44. In the optical path between the lens 25 and the lens 44, a primary image plane F1 optically conjugate with the front focal plane (object plane) of the objective lens 36 is formed. A primary image (eg, an intermediate image) of the sample S is formed on the primary image plane F1.

The light that has passed through the lens 44 is reflected by the mirror 45, passes through the lens 46, and enters the beam splitter 47. The beam splitter 47 has a characteristic that a part of the light incident from the lens 46 is transmitted and a part of the light incident from the lens 46 is reflected. The ratio between the amount of light reflected by the beam splitter 47 and the amount of light transmitted through the beam splitter 47 is arbitrarily set. For example, the ratio may be 8: 2 or any other ratio. The light reflected by the beam splitter 47 enters the lens 48. The lens 48 guides the light reflected by the beam splitter 47 to the image sensor 61. The lens 44, the lens 46, and the lens 48 are, for example, a relay optical system, and form a secondary image surface F2 that is optically conjugate with the primary image surface F1. A secondary image (eg, final image) of the sample S is formed on the secondary image plane F2.

The image sensor 61 is disposed at or near the position of the secondary image plane F2, and captures a secondary image of the sample S. For example, the user can observe the sample S from the image captured by the image sensor 61. However, whether or not the image sensor 61 is arranged is arbitrary, and the image sensor 61 may not be provided.

The imaging optical system 40b guides light (observation light) from the sample S to the user's viewpoint VP. The imaging optical system 40b is configured using elements from the objective lens 36 to the beam splitter 47 of the imaging optical system 40a. The imaging optical system 40 b includes a beam splitter 49 of the imaging optical system 40 c described later on the transmission side of the light incident on the beam splitter 47. The light transmitted through the beam splitter 47 reaches the beam splitter 49. The light transmitted through the beam splitter 49 passes through the lens 52, is reflected by the mirror 53, and enters the eyepiece lens 17. The lens 44, the lens 46, and the lens 52 are, for example, a relay optical system, and a secondary image surface F3 that is optically conjugate with the primary image surface F1 is formed. A secondary image of the sample S is formed on the secondary image plane F3.

The user can observe the secondary image of the sample S through the eyepiece 17. The eyepiece 17 is disposed on the front side (see FIG. 2) of the microscope apparatus 100, for example. For example, the user observes the sample S on the front side of the microscope apparatus 100 and performs operations such as exchanging and moving the sample S from this position.

It should be noted that at least a part of the imaging optical system 40 (imaging optical system 40a and imaging optical system 40b) is not limited to the configuration of FIG. 3, and can be changed as appropriate. For example, the microscope apparatus 100 may not include the imaging optical system 40a or the imaging optical system 40b.

The imaging optical system 40 c guides light (observation light) from the sample S to the imaging unit 60. The imaging optical system 40 c includes a beam splitter 49, a lens 50, and a shutter unit 51. The light (observation light) from the sample S is guided to the beam splitter 49 of the imaging optical system 40c through the elements from the objective lens 36 of the imaging optical system 40a to the beam splitter 47. The imaging optical system 40 c includes a lens 50 and an imaging element 61 of the imaging unit 60 on the light reflection side by the beam splitter 49. The light transmitted through the beam splitter 49 enters the eyepiece 17 through the lens 52 as described above.

The beam splitter 49, the lens 50, and the shutter unit 51 are accommodated in, for example, the lens barrel base 16a (see FIG. 2). The imaging unit 60 having the imaging element 62 is attached in a state of protruding to the −Y side of the lens barrel base 16a. The imaging unit 60 is a so-called camera unit, and is detachably fixed to the lens barrel base 16a. The beam splitter 49 has a characteristic that a part of the light incident from the imaging optical system 40a is transmitted and a part of the light incident from the imaging optical system 40a is reflected. The ratio between the amount of light reflected by the beam splitter 49 and the amount of light transmitted through the beam splitter 49 is arbitrarily set. For example, the ratio may be 8: 2 or any other ratio.

The light reflected by the beam splitter 49 enters the lens 50. The lens 50 guides the light reflected by the beam splitter 49 to the image sensor 62. The lens 50 is, for example, a relay optical system together with the lens 44, the lens 46, and the lens 48, and a secondary image surface F4 optically conjugate with the primary image surface F1 is formed on the imaging element 62. A secondary image of the sample S is formed on the secondary image plane F4. The imaging device 62 of the imaging unit 60 is disposed at or near the position of the secondary image plane F4 and captures a secondary image of the sample S. For example, the user can observe the sample S from the image captured by the image sensor 62.

The shutter unit 51 is disposed so as to be able to be inserted into and retracted from the optical path L on the transmission side of the beam splitter 49. When the shutter unit 51 is inserted into the optical path L, the light transmitted through the beam splitter 49 is blocked by the shutter unit 51. When the shutter 51 is retracted from the optical path L, the light transmitted through the beam splitter 49 is guided to the lens 52 side of the imaging optical system 40b. As described above, the shutter unit 51 restricts the image from reaching the eyepiece 17 on the rear side of the beam splitter 49. The shutter unit 51 is provided on the lens barrel base 16a as described above, and a specific configuration will be described later with reference to another drawing.

Further, as shown in FIG. 3, the imaging optical system 40 a includes a belt run lens 43. The Bertrand lens 43 is a pupil observation optical element. The Bertrand lens 43 is disposed inside the base 11 and is disposed so as to be able to be inserted and retracted with respect to a position in the optical path L that connects the pupil image of the objective lens 36 to the field of view (for example, the image sensor 62). A specific configuration of the Bertrand lens 43 will be described later with reference to another drawing. When the belt run lens 43 is disposed in the optical path L, an image of the pupil conjugate plane (hereinafter referred to as a pupil image) is formed on the secondary image plane F4 (including the secondary image planes F2 and F3). Therefore, the imaging element 62 of the imaging unit 60 can capture a pupil image when the belt run lens 43 is disposed in the optical path L.

By imaging this pupil image with the imaging unit 60, an image formed by each optical element that is a component of the first illumination optical system 20, the second illumination optical system 30, or the imaging optical system 40 is obtained. Further, by retracting the belt run lens 43, an image of the sample surface can be captured by the imaging unit 60. The user can, for example, insert the belt run lens 43 and observe the optical element arranged on the plane conjugate with the pupil plane from the captured image of the pupil image by the image pickup element 62, and retract the belt run lens 43. An optical element arranged on a surface conjugate with the sample surface can be observed from the captured image of the sample surface. In the present embodiment, the image conjugate plane and the plane conjugate with the sample plane are imaged by the imaging device 62, but the present invention is not limited to this, and the image is a plane deviated from the pupil conjugate plane or conjugate with the sample plane. An image of a surface shifted from the surface may be captured by the image sensor 62.

Returning to FIG. 1, the processing device 80 includes an input device 81, a display device 82, an image processing unit 83, a calculation unit 84, and a display control unit 85. The processing device 80 is configured to include a CPU or a memory, and for example, a personal computer is used. Further, the processing device 80 may be formed as a separate device from the microscope main body 10, or may be formed inside the microscope main body 10, for example, on the substrate M in the base 11. In the present embodiment, a correction unit is provided that corrects the position of the component that constitutes one of the optical systems based on the information of the image output from the imaging unit 60. The calculation unit 84 may be a part of the correction unit. In the present embodiment, the calculation unit 84 may not be provided.

The input device 81 can input operation information of the processing device 80. As the input device 81, for example, various devices and devices capable of inputting information such as a mouse, a lever, a touch panel, and a keyboard are used. The input device 81 can input information for adjusting the position and rotational position of optical elements such as the phase ring 26, the polarizer 28, and the analyzer 37, for example. The display device 82 displays information processed by the processing device 80. As the display device 82, for example, various devices and devices such as a liquid crystal panel are used. The display device 82 can display, for example, an image captured by the image sensor 62 of the imaging unit 60. Further, an image captured by the image sensor 61 (see FIG. 3) may be displayed.

The image processing unit 83 performs predetermined image processing such as pattern matching on the image picked up by the image pickup unit 60 (image pickup element 62). For example, the image processing unit 83 extracts an image of an optical element that is a component of the first illumination optical system 20, the second illumination optical system 30, or the imaging optical system 40 from the image captured by the image sensor 62. Is possible.

The calculation unit 84 performs various calculation processes. For example, the calculation unit 84 calculates the difference between the position of the component image extracted by the image processing unit 83 and the target position of the component. The computing unit 84 calculates at least one of the direction and the movement amount for moving the component to the target position. The processing device 80 may store data related to the target position of the component in a storage unit or the like in advance. Further, the processing device 80 may include a communication unit, and may obtain data related to the target position of the component from an external control device or the like via the communication unit.

The display control unit 85 controls the display of the display device 82. For example, the display control unit 85 causes the display device 82 to display the target position of the component extracted by the image processing unit 83 on the image captured by the image sensor 62. The display control unit 85 may display the target position at a luminance that increases the contrast with respect to the image, may blink the target position, or may display the target position in a color different from the main color of the image. .

The display control unit 85 displays the calculation result of the calculation unit 84 on the display device 82. The display control unit 85 causes the display device 82 to display information such as the direction or movement amount calculated by the calculation unit 84 together with the image of the target component. The direction calculated by the calculation unit 84 may be displayed by an arrow as a movement direction instruction mark on the display device 82, for example. For example, the movement amount calculated by the calculation unit 84 may be displayed as a numerical value on a part of the screen (eg, near the target position). Further, when the difference from the target position calculated by the calculation unit 84 is equal to or less than a predetermined threshold value, the processing device 80 may cause the display control unit 85 to display a display corresponding to the necessity of adjustment on the display device 82.

4A and 4B are perspective views showing an example of the lens barrel base 16a, where FIG. 4A is a diagram showing an appearance, and FIG. 4B is a diagram showing an internal configuration. As shown in FIG. 4A, the lens barrel base 16 a has a body 63. The body 63 has a base connection part 63a and a lens barrel mounting part 63b. The base connection part 63 a is connected to the upper surface on the + X side (user side) with respect to the central portion of the base 11. The lens barrel base 16a has a base connecting portion 63a installed on the upper surface of the base 11, and is fixed to the base 11 in a removable state by a fastening member (not shown) such as a fixing pin. Further, the lens barrel mounting portion 63b has the lens barrel 16 mounted thereon, and fixes the lens barrel 16 to the lens barrel base 16a in a removable state by a fastening member (not shown) such as a fixing pin. The eyepiece 17 is instructed to the base 11 through the lens barrel base 16 a and the lens barrel 16.

The body 63 includes an imaging unit 60 having an imaging element 62 and a support unit 12a that fixes the supported portion 15a of the stage 15 described above. The imaging unit 60 is fixed in a state of protruding from the body 63, but may be arranged inside the body 63. In addition, the body 63 includes a manual switching knob 64 and a shutter operation unit 65. The manual switching knob 64 is connected to the shutter operation unit 65 and is used for switching (opening and closing) the shutter unit 51 manually by the user. Therefore, the manual switching knob 64 is formed in a shape that is easy for the user to hold. The manual switching knob 64 is disposed below or near the eyepiece 17.

As shown in FIG. 4B, the shutter operation section 65 has a unit body 65a. The unit body 65a is fixed to the lens barrel base 16a. The beam splitter 49 (see FIG. 3) is fixed to the unit body 65a. The shutter operation unit 65 includes a shutter body 66 and a guide shaft 67. The shutter body 66 is formed integrally with the shutter unit 51.

The shutter body 66 is movable along the guide shaft 67. The guide shaft 67 is fixed to the unit body 65a. The guide shaft 67 is provided in a cylindrical shape or a columnar shape whose outer periphery is circular, but is not limited to this, and may have other shapes. The manual switching knob 64 is attached to the end of the turning shaft 64a. The pivot shaft 64a is rotatably supported by a part of the unit body 65a. The operation of the shutter operation unit 65 and the manual switching knob 64 will be described with reference to FIGS.

5 and 6 are diagrams showing an example of the operation of the shutter operation unit 65, respectively. FIG. 5 and FIG. 6A are perspective views, and FIG. 5 and FIG. FIG. 5A and 5B show a state where the shutter unit 51 is retracted from the optical path, and FIGS. 6A and 6B show a state where the shutter unit 51 is inserted into the optical path.

The shutter operation unit 65 includes an arm 68 (see FIGS. 5B and 6B) and an interlock switch 69 (see FIGS. 5A and 6A). The arm 68 is connected to the turning shaft 64 a of the manual switching knob 64. The arm 68 can turn integrally with the turning shaft 64a in the direction around the turning shaft 64a. The arm 68 has a notch 68a in the longitudinal direction from the distal end side. The width of the notch 68 a is formed substantially constant over the longitudinal direction of the arm 68. A protrusion 66a provided on the shutter body 66 is inserted into the notch 68a.

The guide shaft 67 has recesses 67a and 67b (see FIGS. 5B and 6B). Each of the concave portions 67a and 67b is formed in a shape into which a convex portion 66c provided in the shutter body 66 enters. The protruding portion 66c is held in a state of protruding by an elastic body (not shown) and can be immersed. The shutter body 66 is positioned in the longitudinal direction of the guide shaft 67 by the projection 66c entering either of the recesses 67a and 67b. The recess 67a is provided at a position where the shutter 51 is retracted from the optical path L. The recess 67b is provided at a position where the shutter portion 51 is inserted into the optical path L.

The interlock switch 69 is set so as to operate in a state where the convex portion 66c enters the concave portion 67a or the concave portion 67b of the guide shaft 67. For example, when the convex portion 66 c is in the concave portion 67 a, the shutter portion 51 releases the optical path L and an image is guided to the eyepiece lens 17. In this state, the user can look through the eyepiece lens 17 and, for example, perform an interlock such that the laser light emitted from the light source device 21 (see FIG. 3 and the like) does not reach the eyepiece lens 17. It may be.

The opening / closing operation of the shutter unit 51 will be described. When the user rotates the manual switching knob 64 counterclockwise in the drawing from the state shown in FIGS. 5A and 5B, the turning shaft 64a and the arm 68 similarly turn counterclockwise in the drawing. By this turning of the arm 68, the projection 66a sandwiched between the notches 68a is pulled in the left direction in the figure, the convex part 66c is disengaged from the concave part 67a, and the shutter body 66 is moved along the guide shaft 67 to the left in the figure. Move in the direction.

When the shutter body 66 moves to the left in the drawing and the convex portion 66c enters the concave portion 67b, the shutter portion 51 is inserted into the optical path L as shown in FIGS. The shutter body 66 is positioned with respect to the unit body 65a. In this state, the interlock switch 69 may be turned on. Further, since the shutter unit 51 closes the optical path L, the image cannot be viewed even if the user looks into the eyepiece 17.

Further, from the state shown in FIGS. 6A and 6B, when the user rotates the manual switching knob 64 clockwise in the drawing, the turning shaft 64a and the arm 68 similarly turn clockwise in the drawing. By this turning of the arm 68, the projection 66a sandwiched between the notches 68a is pulled in the right direction in the figure, the convex part 66c is disengaged from the concave part 67b, and the shutter body 66 is moved along the guide shaft 67 to the right in the figure. Move in the direction. When the shutter body 66 moves rightward in the drawing and the convex portion 66c enters the concave portion 67b, the shutter portion 51 returns to the state of being retracted from the optical path L as shown in FIGS. The shutter body 66 is positioned with respect to the unit body 65a.

Although the configuration shown in FIGS. 5 and 6 describes the case where the user manually opens and closes the shutter unit 51, the configuration is not limited to this configuration. For example, the turning shaft 64a may be rotated by a drive source such as an electric motor. In this case, the user may open and close the shutter unit 51 by operating a switch such as an electric motor in order to open and close the shutter unit 51.

7 to 9 are diagrams showing an example of the internal configuration of the base 11. FIG. 7 is a diagram of the internal configuration viewed from the bottom side, and FIG. 8 is a diagram of the internal configuration viewed from the side. 7 and 8 show a state where the belt run lens 43 is inserted in the optical path L. As shown in FIGS. 7 and 8, a support substrate 70 is disposed inside the base 11. The support substrate 70 is disposed in parallel to the XY plane, for example. The support substrate 70 supports the mirror 42, the belt run lens 43, the lens 44, and the mirror 45. The mirror 42, the belt run lens 43, the lens 44, and the mirror 45 are disposed on the back surface side (−Z side) of the support substrate 70.

Also, a belt run lens drive mechanism (element operation unit) 71 is attached to the support substrate 70. The Bertrand lens drive mechanism 71 includes an optical path insertion / removal handle 72, an X position adjustment handle 73, a first shaft 74, a first connecting portion 75, a second shaft 76, and a second connecting portion 77. The optical path insertion / removal handle 72 protrudes outside the microscope body 10 from the + X side of the base 11 (see FIG. 2). The optical path insertion / removal handle 72 is rotatable in the direction around the axis X1 parallel to the X axis, for example. The optical path insertion / removal handle 72 has a protrusion 72a protruding in the radial direction. The user can determine the rotational position of the optical path insertion / removal handle 72 by checking the protruding direction of the protrusion 72a.

The first shaft 74 is disposed along the X direction. The first shaft 74 is integrated with the optical path insertion / removal handle 72. The first shaft 74 is formed in a hollow shape such as a cylindrical shape. The first axis 74 shares the axis X1 with the optical path insertion / removal handle 72. The first shaft 74 is supported by bearings 78 and 79 so as to be rotatable around the axis X1. The bearings 78 and 79 are fixed to the support substrate 70 by a fixing member. The first shaft 74 is supported in a state where movement of the axis X1 in the axial direction (X direction) is restricted.

The first connecting portion 75 connects the first shaft 74 and the belt run lens 43. The 1st connection part 75 has the shaft end part 75a, the plate-shaped member 75b, the rod-shaped member 75c, and the cover member 75d. The shaft end portion 75 a is integrated with the first shaft 74. The shaft end portion 75a shares the axis X1 with the optical path insertion / removal handle 72 and the first shaft 74. The shaft end portion 75a is rotatable in the direction around the axis X1. The plate-like member 75b is fixed to the shaft end portion 75a. When the shaft end portion 75a rotates in the direction around the axis X1, the plate-like member 75b rotates in the direction around the axis X1.

The rod-like member 75c is fixed to the surface on the −X side of the plate-like member 75b. The rod-like member 75c is arranged extending in the −X direction from a position away from the axis X1. The cover member 75d accommodates the belt run lens 43 therein. The cover member 75d has a guide hole 75e on the −Z side surface. The guide hole 75e is a long hole extending in the X direction. The guide hole 75e is provided through the inside and outside of the cover member 75d. Further, the cover member 75d is movable in the Y direction by the guide portion 75f.

The X position adjustment handle 73 is disposed inside the optical path insertion / removal handle 72 and is provided so as to protrude from the optical path insertion / removal handle 72 in the + X direction. The X position adjustment handle 73 protrudes from the + X side of the base 11 together with the optical path insertion / removal handle 72 to the outside of the microscope body 10. The X position adjustment handle 73 can move in the X direction independently of the optical path insertion / removal handle 72 and can rotate about the axis X1. Further, a part of the X position adjusting handle 73 is screw-coupled to a screw portion provided on the inner periphery of the + X side end of a fixing portion 70a (see FIG. 8) attached to the support substrate 70.

The second shaft 76 is arranged along the X direction. The second shaft 76 is integrated with the X position adjusting handle 73. The second shaft 76 is formed in a columnar shape or a cylindrical shape, for example. The second shaft 76 is disposed inside the first shaft 74. The second shaft 76 is rotatable relative to the first shaft 74 in the direction around the axis X1. When the X position adjusting handle 73 is rotated, the X position adjusting handle 73 (second shaft 76) is moved in the X direction with respect to the optical path inserting / removing handle 72 (first shaft 74) by screw connection with the fixing portion 70a. Move relative to. The second shaft 76 is disposed through the plate-like member 75b.

The second connecting portion 77 connects the second shaft 76 and the belt run lens 43. The 2nd connection part 77 has the plate-shaped member 77a and the lens holding part 77b. The plate-like member 77a is attached to the end surface on the −X side of the lens holding portion 77b, and is formed so as to sandwich a part of the outer periphery of the rod-like member 75c. The −X side end of the second shaft 76 is in contact with the plate member 77. Since the plate-like member 77a sandwiches the outer periphery of a part of the rod-like member 75c as described above, the plate-like member 77a can move in the X direction with respect to the rod-like member 75c. When rotating around the axis X1, it moves in the Y direction along the guide portion 75f.

The lens holding portion 77b holds the belt run lens 43. The lens holding portion 77b is fixed to the surface on the −X side of the plate-like member 77a. The lens holding portion 77b is given an elastic force in the + X direction by an elastic member (not shown) such as a spring in the cover member 75d. When the plate-like member 77a moves in the + X direction, the lens holding portion 77b moves in the + X direction by the elastic force of the elastic member. Further, when the plate-like member 77a moves in the −X direction, it moves in the −X direction against the elastic force of the elastic member. The lens holding portion 77b is relatively movable in the X direction with the cover member 75d. The lens holding portion 77b has a protrusion 77c on the −Z side. The protrusion 77c is disposed in the guide hole 75e of the cover member 75d. When the lens holding portion 77b moves in the X direction relative to the cover member 75d, the protrusion 77c is guided in the X direction by the guide hole 75e. For this reason, the lens holding part 77b is movable along the X direction. Further, since the protrusion 77c is disposed in the guide hole 75e, relative rotation between the cover member 75d and the lens holding portion 77b is restricted.

Further, when the X position adjusting handle 73 is rotated, the X position adjusting handle 73 moves in the X direction by screw connection with the fixing portion 70a (see FIG. 8). Thereby, the lens holding part 77b can be moved relative to the cover member 75d in the X direction via the second shaft 76 and the plate-like member 77a. That is, by rotating the X position adjustment handle 73, the belt run lens 43 held by the lens holding portion 77b can be moved along the optical path L parallel to the X axis.

The belt run lens driving mechanism 71 can insert the belt run lens 43 into the optical path L or retract it from the optical path L by rotating the optical path insertion / removal handle 72 in the direction around the axis X1. FIG. 9A shows a state where the belt run lens 43 is inserted into the optical path L, and FIG. 9B shows a state where the belt run lens 43 is retracted from the optical path L. As shown in FIG. 9A, in a state where the belt run lens 43 is inserted in the optical path L, the belt run lens 43 is retracted from the optical path L by rotating the optical path insertion / removal handle 72 around the axis X1. It is possible.

9A, the optical path insertion / removal handle 72 is rotated in the direction around the axis X1 (counterclockwise direction) when viewed from the + X side. By the rotation of the optical path insertion / removal handle 72, the first shaft 74, the shaft end portion 75a, and the plate member 75b are integrally rotated in the direction around the axis X1. The rod-like member 75c moves around the axis X1 by the rotation of the plate-like member 75b. By the movement of the rod-shaped member 75c, as shown in FIG. 9B, the cover member 75d and the lens holding portion 77b are guided by the guide portion 75f and moved in the −Y direction. The belt run lens 43 is retracted from the optical path L by the movement of the lens holding portion 77b.

9B, the belt run lens 43 is inserted into the optical path L by rotating the optical path insertion / removal handle 72 in the direction around the axis X1 from the state where the belt run lens 43 is retracted from the optical path L as shown in FIG. It is possible. In this case, the optical path insertion / removal handle 72 is rotated in the direction around the axis X1 (clockwise direction) when viewed from the + X side. By the rotation of the optical path insertion / removal handle 72, the first shaft 74, the shaft end portion 75a, and the plate member 75b are integrally rotated in the direction around the axis X1. The rod-like member 75c moves around the axis X1 by the rotation of the plate-like member 75b. By the movement of the rod-shaped member 75c, the cover member 75d and the lens holding portion 77b are guided by the guide portion 75f and moved in the + Y direction. The belt run lens 43 is inserted into the optical path L by the movement of the lens holding portion 77b.

Further, when the belt run lens 43 is inserted in the optical path L (state shown in FIG. 9A), the X position adjustment handle 73 is rotated around the axis X1 so that the X direction relative to the optical path insertion / removal handle 72 is obtained. The relative position of changes. For example, when the X position adjustment handle 73 is rotated clockwise as viewed from the + X direction, the end of the second shaft 76 pushes the plate-like member 77a in the −X direction and moves it. Due to the movement of the plate member 77a, the lens holding portion 77b moves in the −X direction with respect to the cover member 75d against the elastic force of the elastic member. The belt run lens 43 moves in the −X direction by the movement of the lens holding portion 77b. For this reason, the belt run lens 43 moves away from the mirror 42 and closer to the lens 44.

Also, for example, when the X position adjustment handle 73 is rotated counterclockwise when viewed from the + X direction, the end of the second shaft 76 moves in the + X direction. Accordingly, the plate-like member 77a and the lens holding portion 77b are moved in the + X direction with respect to the cover member 75d by the elastic force of the elastic member. Due to the movement of the lens holding portion 77b, the belt run lens 43 moves in the + X direction. For this reason, the belt run lens 43 moves closer to the mirror 42 and away from the lens 44. Thus, the position of the belt run lens 43 in the optical path L can be adjusted by rotating the X position adjusting handle 73.

7 to 9 describe the case where the user manually inserts and retracts the belt run lens 43 and moves the belt run lens 43 in the optical path L. However, the configuration is not limited to this configuration. For example, the first shaft 74 or the second shaft 76 may be rotated by a drive source such as an electric motor. In this case, the user may operate a switch such as an electric motor to insert and retract the belt run lens 43 and move the belt run lens 43 in the optical path L. As shown in FIGS. 7 and 9, the support substrate 70 includes a switching unit 90 for switching the second objective lens 41 (magnification lens) shown in FIG. 3. For example, the switching unit 90 can rotate a turret (not shown) that holds the plurality of second objective lenses 41 by rotating a knob, and can place the predetermined second objective lens 41 in the optical path L.

Next, an image processing method according to the embodiment will be described. FIG. 10 is a flowchart illustrating an example of an image processing method according to the embodiment. In the present embodiment, using the microscope apparatus 100 described above, a positioning operation (for example, a pupil centering operation) is performed on an optical element that is a component of the first illumination optical system 20 or the imaging optical system 40. The operation in this case will be described.
In this embodiment, the means for performing the alignment operation is referred to as a correction unit. The operation of the correction unit may be performed manually by the user, or may be performed automatically based on image information provided with a correction operation device that drives the operation unit.
Based on the information of the image output from the imaging unit 60, the correction unit positions the components constituting any one of the optical systems (the first illumination optical system 20, the second illumination optical system 30, and the imaging optical system 40). Correct. The correction unit corrects the position of the component based on the image information and the target information of the predetermined component. The image information is image position information, and the target information is target position information.

The processing device 80 may display a menu image for selecting an adjustment target on the display device 82 prior to imaging by the imaging device 62 in step ST01. On the menu screen displayed on the display device 82, for example, the names of optical elements to be adjusted are displayed side by side, and by selecting one with the input device 81 such as a mouse or a keyboard, the object to be adjusted is selected. May be determined.

In addition, after determining the adjustment target, the processing device 80 determines whether or not a pupil image is necessary when adjusting the selected optical element or the like. The Bertrand lens driving mechanism 71 may be operated to display on the display device 82 such that the belt run lens 43 is inserted into the optical path L. In this case, the processing device 80 may cause the display device 82 to display the operation method of the belt run lens driving mechanism 71. For example, the processing device 80 may display an image of the optical path insertion / removal handle 72 on the display device 82 and display an arrow indicating the rotation direction in an overlapping manner. The user can easily insert the belt run lens 43 into the optical path L by checking the screen of the display device 82.

Further, the processing device 80 may confirm that the belt run lens 43 is properly inserted into the optical path L by using a sensor (not shown) or the like, and display the insertion completion of the belt run lens 43 on the display device 82. When the belt run lens 43 is inserted and retracted by a drive source such as an electric motor, the processing device 80 automatically drives the drive source to insert the belt run lens 43 when the user selects an adjustment target. May be.

Further, similarly to the insertion of the belt run lens 43, the processing device 80 may display on the display device 82 so that the user operates the manual switching knob 64 to insert the shutter portion 51 into the optical path L. In this case, the processing device 80 may display the operation method of the manual switching knob 64 on the display device 82. For example, the processing device 80 may display an image of the manual switching knob 64 on the display device 82 and display an arrow indicating the rotation direction in an overlapping manner. The user can easily insert the shutter unit 51 into the optical path L by checking the screen of the display device 82.

Further, the processing device 80 may confirm that the shutter unit 51 is properly inserted into the optical path L with a sensor (not shown) or the like, and display the completion of insertion of the shutter unit 51 on the display device 82. When the shutter unit 51 is opened and closed (inserted and retracted) by a driving source such as an electric motor, the processing device 80 drives the driving source when the user selects an adjustment target or inserts the belt run lens 43. Then, the shutter unit 51 may be automatically inserted.

Subsequently, as illustrated in FIG. 10, the processing device 80 causes the image sensor 62 to capture a pupil image of a predetermined optical element (step ST01). The processing device 80 irradiates light from the light source device 21 of the first illumination optical system 20 and images the pupil image formed at the position of the secondary image plane F4 by the imaging element 62. The processing device 80 may irradiate light from the light source device 21 after confirming that the belt run lens 43 is inserted in the optical path L and that the shutter unit 51 is inserted in the optical path L.

Next, the image processing unit 83 extracts a feature point of a component (for example, a target optical element) from the image captured by the image sensor 62 (step ST02). In step ST02, the image processing unit 83 extracts feature points of the target optical element by pattern matching or the like using the captured image and, for example, a plurality of types of shapes of the optical element stored in the storage unit or the like. Good. The feature points include, for example, the shape of some or all of the components such as the light source device or the optical element, the luminance based on the components, the shape to be imaged in the vicinity of the imaging target, and the like.

Next, the calculation unit 84 calculates the difference between the feature point extracted by the image processing unit 83 and the target (step ST03). In step ST03, for example, the difference between the position of the optical element from which the feature point is extracted and the target position of the optical element is calculated. Subsequently, the calculation unit 84 determines whether or not the difference calculated in step ST03 is appropriate (step ST04). “Appropriate” means, for example, that the optical element is arranged at the target position (the difference is equal to or less than the threshold value), or that there is no unnecessary state (for example, bubbles or foreign matter) in the vicinity of the imaging target. .

If the difference calculated in step ST04 is not appropriate (NO in step ST04), the display control unit 85 causes the display device 82 to display assist information superimposed on the captured image (step ST05). In step ST03, the calculation unit 84 may calculate at least one of a direction and a movement amount for moving the target optical element to the target position. For example, the calculation unit 84 calculates the center (or center of gravity) position of the optical element from the feature point based on the captured image, and compares the calculated position with the target position stored in the storage unit or the like to determine the difference from the target position. Further, one or both of the direction and the amount of movement of the optical element to the target position may be calculated.

Next, the assist information in step ST04 includes, for example, information calculated by the calculation unit 84 such as the direction in which the optical element is moved to the target position and the amount of movement thereof. In step ST <b> 05, for example, the display control unit 85 may display the target positions of the constituent elements extracted by the image processing unit 83 on the display device 82 so as to overlap the image. Further, the display control unit 85 may cause the display device 82 to display the direction calculated by the calculation unit 84 (the direction in which the optical element is moved to the target position) or the movement amount as a movement direction instruction mark so as to overlap the image with an arrow. Good. Subsequently, the user moves the optical element or the like according to the assist information in step ST04 (step ST06). The user can easily adjust the position of the optical element by operating the optical element while checking the image displayed on the display device 82 and the assist information. When the optical element can be moved by a drive source such as an electric motor, the optical element may be moved by driving the drive source instead of being operated by the user.

After step ST06, the process returns to step ST01, and the above-described steps ST01 to ST04 are repeated. In step ST01, the imaging unit 60 may display the moving state of the optical element on the display device 82 in real time by imaging a moving image or every several microseconds. In addition, an image captured by the imaging unit 60 may be displayed on the display device 82 as appropriate by the user operating a button or the like.

Further, when the difference calculated in step ST04 is appropriate (YES in step ST04), the processing device 80 displays on the display device 82 that the difference is appropriate (step ST07). For example, the processing device 80 displays an indication of appropriateness (for example, adjustment) when the difference calculated by the calculation unit 84 in step ST03 is equal to or smaller than a predetermined threshold value, or when there are no bubbles or the like in the observation target and its vicinity. The display control unit 85 may display on the display device 82 a display corresponding to unnecessary or completion of adjustment. The user can easily confirm that there is no need to move the target optical element or that there are no bubbles or the like in the observation target and its vicinity by checking the display indicating that adjustment is unnecessary.

Note that the optical elements need to be adjusted differently when the observation method using the microscope body 10 is different. For example, the processing device 80 may display a plurality of observation methods side by side on the initial menu screen and display them on the display device 82. When an observation method is selected from this menu, the processing device 80 displays optical elements that require adjustment on the display device 82. It may be displayed. Thereby, the user can easily recognize an optical element necessary for the selected observation method, and can manually insert the optical element into the optical path L. When optical elements necessary for the selected observation method are inserted into and retracted from the optical path L by a drive source such as an electric motor, the processing device 80 drives the drive source when the observation method is selected from the menu. Then, the drive source may be controlled so that an optical element necessary for the observation method is automatically inserted into the optical path L.

After the adjustment of the optical element as described above is completed, the sample S is placed on the stage 15 (see FIG. 2), and the sample S is observed. When observing the sample S, both the belt run lens 43 and the shutter unit 51 are retracted from the optical path L. When the adjustment of the optical element described above is completed, the processing device 80 may cause the display device 82 to display a display corresponding to the observation of the sample S. Further, when the processing device 80 performs the operations of the belt run lens 43 and the shutter unit 51 by the drive source, after the adjustment of the optical element is completed, the drive source is driven to move both the belt run lens 43 and the shutter unit 51 to the optical path L. May be automatically evacuated.

11 to 14 show examples displayed by the display device 82. FIG. 11 is a diagram illustrating an example of display contents of the display device 82 when the position of the phase ring 26 is adjusted. In FIG. 11, an image captured by the image sensor 62 with both the belt run lens 43 and the shutter unit 51 inserted in the optical path L is used. First, the processing device 80 extracts the position of the phase film of the objective lens 36. The position of the phase film is the target position of the phase ring 26. The processing device 80 irradiates light from the light source device 21 in a state where the phase ring 26 is retracted from the optical path L, and causes the imaging device 62 to perform imaging. As shown in FIG. 11A, the display control unit 85 causes the display device 82 to display the phase film image 17a as the imaging result. Thereafter, the image processing unit 83 extracts the position of the phase film based on the imaging result. Note that the processing device 80 may extract the position of the phase film based on position information stored in advance in a storage unit or the like.

Next, the processing device 80 irradiates light from the light source device 21 with the phase ring 26 inserted in the optical path L, and takes an image of the phase ring 26 by the imaging element 62. For example, in the example shown in FIG. 11B, the phase film image 17a and the phase ring image 26a are displayed on the display device 82 in a state where they overlap each other. The image processing unit 83 extracts the position (feature point) of the phase ring based on the phase ring image 26a.

Next, the calculation unit 84 calculates the difference between the extracted position of the phase ring 26 and the position of the phase film that is the target position. When the difference is greater than or equal to the threshold (when the difference is not appropriate), the display control unit 85 uses the calculation result calculated by the calculation unit 84 as the assist information, for example, as shown in FIG. The direction in which the ring 26 is moved to the target position is displayed on the display device 82 by an arrow (movement direction instruction mark) 91. Thus, the user can easily adjust the phase ring 26 to the target position by moving the phase ring 26 in the direction of the arrow 91 displayed on the display device 82. Note that the target position (phase film) may be difficult to see depending on the magnification or the like. For example, the target position may be highlighted.

Note that when the user moves the phase ring 26, the situation may be continuously captured by the image sensor 62 and displayed on the display device 82 as a moving image. Thus, the user can easily confirm whether the direction or amount of movement of the phase ring 26 is correct while viewing the display device 82. In addition, the calculation unit 84 may perform the above calculation for each image picked up by the image sensor 62, and may continue to display the arrow 91 on the display device 82 when the calculation result is equal to or greater than a predetermined threshold value.

In addition, when the difference between the position of the phase ring 26 and the target position is equal to or smaller than a predetermined threshold (when the difference is appropriate), the processing device 80 determines that the difference is appropriate and does not require adjustment (or adjustment is completed). For example, a circular display 92 as shown in FIG. 11D may be displayed on the display device 82 by the display control unit 85. The display 92 may be displayed colored in green or the like. As a result, the user can confirm that the phase ring 26 is arranged at the substantially target position. When the phase ring 26 is already in the vicinity of the target position before the user adjusts the phase ring 26 (when the difference between the position of the phase ring 26 and the target position is equal to or smaller than a predetermined threshold value), FIG. Is displayed on the display device 82 from the beginning. Thus, the user can easily confirm that the phase ring 26 does not require adjustment.

FIG. 12 is a diagram showing an example of display contents of the display device 82 when adjusting the rotation positions of the polarizer 28 and the analyzer 37. In FIG. 12, an image captured by the image sensor 62 with both the belt run lens 43 and the shutter unit 51 inserted in the optical path L is used. First, the processing device 80 irradiates light from the light source device 21 in a state where the polarizer 28 and the analyzer 37 are inserted in the optical path L, and performs imaging by the imaging element 62. For example, as illustrated in FIG. 12A, an image 93 that is an imaging result is displayed on the display device 82. The calculation unit 84 may calculate the difference from the target from the luminance (feature point) of the image 93.

When the difference is not appropriate, that is, when the luminance is larger than a predetermined threshold, for example, the rotation direction of the polarizer 28 may be displayed on the display device 82 as assist information. When the user rotates the polarizer 28 from the state shown in FIG. 12A, the luminance of the image 93 changes as shown in FIG. The user rotates the polarizer 28 while confirming the luminance of the image 93 by the display device 82, and adjusts the rotation position of the polarizer 28 so that the luminance of the image 93 is the lowest, as shown in FIG. When the polarizer 28 and the analyzer 37 have a crossed Nicols relationship, the image 93 is the darkest (the luminance is the lowest).

The processing device 80 determines, for example, whether or not the luminance value of a part of the image (for example, the central portion) is equal to or less than a predetermined threshold, or whether the luminance value is the minimum value, by the arithmetic unit 84. Also good. In addition, as a result of the determination by the calculation unit 84, when the luminance of a part of the image described above is equal to or lower than a predetermined threshold value, or when the luminance value becomes the minimum value (that is, when the difference is appropriate), processing is performed. The device 80 may be displayed on the display device 82 by the display control unit 85 with a display corresponding to no adjustment (or adjustment completed), for example, a circular display 94 as shown in FIG. Good. The display 94 may be colored and displayed. Thereby, it is possible to notify the user that the polarizer 28 is disposed at an appropriate position. In the above description, the user rotates the polarizer 28, but the analyzer 37 may be rotated.

FIG. 13 is a diagram showing an example of display contents of the display device 82 when bubbles are extracted. In FIG. 13, an image captured by the image sensor 62 in a state where both the belt run lens 43 and the shutter unit 51 are inserted in the optical path L is used. For example, when immersion liquid is supplied between the objective lens 36 and the sample S, the processing device 80 irradiates light from the light source device 21 and performs imaging with the imaging element 62. As a result, for example, the image shown in FIG. 13A is captured and displayed on the display device 82.

The image processing unit 83 detects bubbles that are feature points from the captured image. For example, the bubble may be detected by dividing the image into a plurality of images, obtaining the luminance for each of the images, and detecting that there is a bubble in the segment having the luminance corresponding to the bubble, or detecting the bubble by pattern matching. Good. When a bubble is detected (when the difference from the target is not appropriate), for example, as shown in FIG. 13B, the display control unit 85 displays a display 96 in the area corresponding to the bubble as assist information. May be. The display 96 may be displayed in red or the like. In addition, the calculation unit 84 may calculate the size or number of bubbles and perform pass / fail determination in the observation of the sample S. The user can easily confirm the presence of bubbles by looking at the image on the display device 82. In addition, when the user determines that the bubbles interfere with the observation of the material S, the user may remove the bubbles, such as replacing the immersion liquid.

Further, the image shown in FIG. 13 is picked up by the image pickup device 62 while changing the focus position of the pupil. Bubble detection is performed in the same manner as described above based on an image obtained by changing the focus position of the pupil. The processing device 80 may change the focus position of the pupil with a drive source (not shown). When the focus position of the pupil is changed manually by the user, the user may take an image for each changed focus position, or the processing device 80 may take an image. The change width of the focus position may correspond to the thickness of the immersion liquid.

FIG. 14 is a diagram showing an example of display contents of the display device 82 when the center of the field stop image and the center of the optical axis are matched. In FIG. 14, an image captured by the image sensor 62 with both the belt run lens 43 and the shutter unit 51 retracted from the optical path L is used. The field stop image looks at the image on the sample surface (sample surface), not the pupil conjugate surface. First, the processing device 80 irradiates light from the light source device 21 and performs imaging with the imaging element 62. For example, as shown in FIG. 14A, the display control unit 85 superimposes the picked-up image and uses a field stop image and a cross mark 97 indicating the center of the field stop image as a feature point as assist information. Then, a cross mark 98 indicating the center of the optical axis and a determination mark 99 indicating whether or not both coincide with each other are displayed on the display device 82.

FIG. 14A shows a case where the center of the display screen of the display device 82 and the center of the optical axis coincide with each other. However, the present invention is not limited to this, and the center of the display screen is shifted from the center of the optical axis. May be. The user moves a field stop (not shown) so that the cross mark 97 and the cross mark 98 coincide with each other. Further, the calculation unit 84 calculates, for example, the difference between the cross mark 97 and the cross mark 98, and uses the direction in which the cross mark 97 is moved to the cross mark 98 as a movement direction instruction mark, as in FIG. You may display on the display apparatus 82 by the arrow. Since the cross marks 97 and 98 are displayed on the display device 82, the field stop can be easily adjusted to the target position.

When the cross mark 97 and the cross mark 98 match, the display control unit 85 determines that the difference is appropriate, and displays a display corresponding to adjustment unnecessary (or adjustment completed), for example, as shown in FIG. The mark 99 is displayed thickly. The determination mark 99 may be displayed in a green color or the like. Thereby, the user can easily confirm that the field stop is arranged at the target position.

The microscope apparatus 100 according to the present embodiment extracts an image of an optical element that is a component such as the first illumination optical system 20 or the imaging optical system 40 from an image captured by the imaging element 62, and the image processing unit 83 A display device that calculates the difference between the position of the extracted image of the optical element and the target position of the optical element, displays the direction, etc., so as to grasp the target position and move the optical element to the target position Therefore, the operation depending on the user's sense is reduced, and the optical element or the like can be easily arranged at the target position. Furthermore, in this embodiment, it is also possible to adjust the optical axis of the laser emission end of the fluorescent illumination device used for fluorescence observation.

As mentioned above, although embodiment was described, the technical scope of this invention is not limited to the aspect demonstrated by above-described embodiment etc. In addition, one or more of the requirements described in the above-described embodiments may be omitted. In addition, the requirements described in the above-described embodiments and the like can be combined as appropriate. In addition, as long as it is permitted by law, the disclosure of all the references cited in the above-described embodiments and the like is used as a part of the description of the text.

In the above-described embodiment, the configuration in which the user manually adjusts the position of the optical element after confirming the image displayed on the display device 82 has been described as an example. However, the present invention is not limited to this. For example, when the microscope main body 10 includes a driving device for moving each optical element (component) such as the first illumination optical system 20 or the imaging optical system 40, the processing device 80 calculates the calculation unit 84. Based on the result, a configuration may be adopted in which a driving device corresponding to each optical element is driven to perform automatic adjustment such that each optical element is moved to a target position. Thereby, the user can save the trouble of adjusting the optical element.

In the above-described embodiment, the microscope apparatus 100 may include a mechanism for performing differential interference observation and a mechanism for performing phase difference observation. Further, the microscope apparatus 100 may be a stereomicroscope. The microscope apparatus 100 may be a so-called upright microscope in which the objective lens 36 is disposed above the stage 15.

L ... optical path, 10 ... microscope body, 20 ... first illumination optical system, 30 ... second illumination optical system, 40, 40a, 40b, 40c ... imaging optical system, 43. ... Belt run lens (pupil observation optical element), 60... Imaging unit, 80... Processing device, 82... Display device, 83. Display control unit, 91... Arrow (movement direction instruction mark), 100.

Claims (24)

1. An illumination optical system for irradiating the sample with light;
   An imaging optical system for imaging light from the sample;
   An imaging unit that captures an image;
   A processing device for processing an image output from the imaging unit;
   A microscope apparatus, comprising: a correction unit that corrects the position of a component that constitutes one of the optical systems based on information on the image.
2. 3. The microscope apparatus according to claim 1, wherein the correction unit corrects the position of the component based on the information on the image and target information on a predetermined component.
3. 3. The microscope apparatus according to claim 1, wherein the image information is image position information, and the target information is target position information.
4. 4. The microscope apparatus according to claim 2, wherein the information of the image is a center position of a field stop.
5. The microscope apparatus according to claim 3 or 4, further comprising a calculation unit that calculates a difference between the position of the image and the target position.
6. The microscope apparatus according to claim 5, wherein the calculation unit calculates at least one of a direction and a movement amount for moving the component to a target position.
7. The microscope apparatus according to any one of claims 1 to 6, wherein the treatment apparatus includes a display device that displays the image and a display control unit that controls display of the display device.
8. The microscope apparatus according to claim 7, wherein the display control unit causes the display device to display a target position of the component extracted by the image processing unit together with the image.
9. The microscope apparatus according to any one of claims 5 to 8, wherein the display control unit displays a calculation result of the calculation unit on the display device.
10. 10. The microscope apparatus according to claim 9, wherein the display control unit causes the display device to display a direction in which the component calculated by the calculation unit is moved to a target position by a movement direction instruction mark.
11. The processing device causes the display control unit to display on the display device a display corresponding to adjustment unnecessary when the difference calculated by the calculation unit is equal to or less than a predetermined threshold value. The microscope apparatus according to claim 1.
12. An eyepiece that can visually observe an image of the sample by the imaging optical system;
    The microscope apparatus according to any one of claims 1 to 11, further comprising a shutter unit that is inserted into an optical path in order to restrict an image from reaching the eyepiece lens behind the imaging unit.
13. A lens barrel base for supporting the eyepiece;
    The microscope apparatus according to claim 12, wherein the shutter unit is provided on the lens barrel base.
14. The microscope apparatus according to claim 13, wherein the imaging unit is arranged on the lens barrel base.
15. The microscope apparatus according to any one of claims 12 to 14, further comprising a shutter operation unit that inserts and retracts the shutter unit.
16. The microscope apparatus according to claim 15, wherein the shutter operation unit is provided below or in the vicinity of the eyepiece.
17. The microscope according to claim 12 or 16, wherein when the image processing unit extracts an image of the component, the processing device operates the shutter operation unit to insert the shutter unit into an optical path. apparatus.
18. The microscope apparatus according to any one of claims 1 to 17, wherein a pupil observation optical element for projecting the image of the pupil to the imaging unit is insertable at a position of the imaging optical system or in the vicinity of the pupil. .
19. The microscope apparatus according to claim 18, further comprising an element operation unit that inserts or retracts the pupil observation optical element into or from the optical path.
20. A driving device for moving components constituting the illumination optical system or the imaging optical system;
    The microscope apparatus according to claim 1, wherein the processing device drives the driving device corresponding to the component based on a calculation result of the calculation unit to move the component to a target position.
21. The microscope according to any one of claims 1 to 20, wherein the processing device calculates a difference between a sample or a state in the vicinity of the sample extracted by the image processing unit and a target state by the calculation unit. apparatus.
22. The image processing unit extracts air bubbles existing in the vicinity of the sample or the sample from the image,
    The microscope apparatus according to claim 21, wherein the calculation unit performs pass / fail determination by comparing the size or number of bubbles with respect to the target state.
23. In the microscope apparatus comprising: an illumination optical system that irradiates light onto the sample; an imaging optical system that forms an image of light from the sample; and an imaging unit that captures an image of the sample by the imaging optical system; A method of processing the image captured in
    Extracting an image of a component constituting the illumination optical system or the imaging optical system from the image;
    An image processing method comprising: calculating a difference between the extracted image position of the component and a target position of the component.
24. A processing apparatus of a microscope apparatus comprising: an illumination optical system that irradiates light on a sample; an imaging optical system that forms an image of light from the sample; and an imaging unit that captures an image of the sample by the imaging optical system. And
    An image processing unit that extracts an image of a component constituting the illumination optical system or the imaging optical system from the image captured by the imaging unit;
    A processing apparatus comprising: an arithmetic unit that calculates a difference between a position of an image of the component extracted by the image processing unit and a target position of the component.

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