阅读说明：本技术 扫描型显微镜单元 (Scanning microscope unit ) 是由 山下慈郎 田边康行 松田俊辅 寺田浩敏 于 2020-03-26 设计创作，主要内容包括：实施方式为安装于显微镜(50)的连接端口(P1)的共聚焦显微镜单元(1),包括：光源(10a～10d),其向观察对象的试样(M)输出照射光；光检测器(13a～13d),其检测根据照射光而从试样产生的观察光；扫描镜(4),其使照射光在试样(M)上扫描,并将从试样(M)产生的观察光朝向光检测器(13a～13d)引导；扫描透镜(7),其将由扫描镜(4)扫描的照射光导光至显微镜光学系统,将由显微镜光学系统成像的观察光导光至扫描镜(4)；镜筒(3),其固定有扫描透镜(7)；配件部(21),其用于将镜筒(3)安装于连接端口(P1)；和可动部(22),其以可变更镜筒(3)相对于配件部(21)的角度的方式支撑镜筒(3)。(The embodiment is a confocal microscope unit (1) mounted on a connection port (P1) of a microscope (50), comprising: light sources (10 a-10 d) that output irradiation light to a sample (M) to be observed; photodetectors (13 a-13 d) that detect observation light generated from the sample in response to the irradiation light; a scanning mirror (4) that scans the sample (M) with the irradiation light and guides the observation light generated from the sample (M) to the photodetectors (13 a-13 d); a scanning lens (7) that guides the illumination light scanned by the scanning mirror (4) to the microscope optical system and guides the observation light imaged by the microscope optical system to the scanning mirror (4); a lens barrel (3) to which a scanning lens (7) is fixed; a fitting part (21) for attaching the lens barrel (3) to a connection port (P1); and a movable part (22) that supports the lens barrel (3) so that the angle of the lens barrel (3) relative to the attachment part (21) can be changed.)

1. A scanning microscope unit characterized in that,

is a scanning microscope unit which is mounted on a connection port of a microscope having a microscope optical system to constitute a scanning microscope,

the scanning microscope unit includes:

a light source that outputs irradiation light toward a sample to be observed;

a photodetector that detects observation light generated from the sample in accordance with the irradiation light;

a scanning mirror that scans the irradiation light output from the light source over the sample and guides the observation light generated from the sample according to the irradiation light toward the photodetector;

a scanning lens that guides the illumination light scanned by the scanning mirror to the microscope optical system and guides the observation light imaged by the microscope optical system to the scanning mirror;

a housing to which a scanning lens is fixed;

a fitting portion for mounting the housing to the connection port; and

and a movable portion that supports the housing so that an angle of the housing with respect to the attachment portion can be changed.

2. Scanning microscope unit according to claim 1,

the movable unit can change the angle of the housing so that the optical axis of the scanning lens is parallel to the optical axis of the microscope optical system.

3. Scanning microscope unit according to claim 1 or 2,

the movable portion is configured to be rotatable with respect to the attachment portion.

4. Scanning microscope unit according to claim 3,

the rotation center of the movable portion is included in an image forming surface of the microscope optical system.

5. The scanning microscope unit of any one of claims 1 to 4,

the scanning mirror is a MEMS mirror.

6. Scanning microscope unit according to any of claims 1 to 5,

the photodetector detects fluorescence generated as the observation light by irradiation of the irradiation light.

7. Scanning microscope unit according to any of claims 1 to 5,

the photodetector detects, as the observation light, reflected light generated as a result of irradiation of the irradiation light.

8. Scanning microscope unit according to any of claims 1 to 7,

further comprising: an aperture member that restricts a beam of the observation light returned via the scanning mirror,

the light detector detects the observation light that has passed through the aperture member.

9. Scanning microscope unit according to claim 8,

the aperture part is a pinhole plate.

Technical Field

The present invention relates to a scanning microscope unit constituting a scanning microscope.

Background

Conventionally, a microscope apparatus capable of obtaining an image of a specimen to be observed is known. For example, patent document 1 listed below discloses a microscope connection unit including a microscope connection port connected to a microscope, a stimulation unit that irradiates a specimen with light, an observation unit that detects light emitted from the specimen, and an optical path combining unit that combines optical paths that optically connect the microscope to the stimulation unit and the observation unit. In the microscope connecting unit having such a configuration, for example, the specimen is imaged by irradiating the specimen with excitation light, using fluorescence generated by the irradiation, and detecting the fluorescence.

Documents of the prior art

Patent document

Patent document 1: japanese patent laid-open publication No. 2011-90248

Disclosure of Invention

Technical problem to be solved by the invention

In the conventional microscope connecting unit as described above, when the unit is connected to a microscope of each kind or each manufacturer for use, the positional relationship between the optical axis of the lens in the microscope and the mirror for scanning the excitation light in the microscope connecting unit may be unstable. Therefore, the signal intensity and the resolution of the observed image tend to decrease.

The embodiment has been made in view of the above problems, and it is an object of the invention to provide a scanning microscope unit capable of forming an image with maintained signal intensity and resolution.

Means for solving the problems

One aspect of the present invention is a scanning microscope unit which is attached to a connection port of a microscope having a microscope optical system to constitute a scanning microscope, the scanning microscope unit including: a light source that outputs irradiation light toward a sample to be observed; a photodetector that detects observation light generated from the sample in accordance with the irradiation light; a scanning mirror that scans irradiation light output from the light source on the sample and guides observation light generated from the sample according to the irradiation light to the photodetector; a scanning lens that guides the irradiation light scanned by the scanning mirror to the microscope optical system and guides observation light imaged by the microscope optical system to the scanning mirror; a housing to which a scanning lens is fixed; a fitting portion for attaching the housing to the connection port; and a movable portion that supports the housing so that an angle of the housing with respect to the attachment portion can be changed.

According to the scanning microscope unit of the above aspect, the irradiation light emitted from the light source is scanned on the sample via the scanning mirror, the scanning lens, and the external microscope, and the observation light generated from the sample based on the scanning light is detected by the photodetector via the external microscope, the scanning lens, and the scanning mirror. In the scanning microscope unit, a housing to which the scanning lens is fixed is attached to a connection port of the microscope by a fitting portion, and an angle of the housing with respect to the fitting portion can be changed. With this configuration, the optical axis of the scanning lens can be aligned in the direction of the optical axis of the optical system of the microscope in accordance with the microscope to be mounted. As a result, it is possible to realize an image in which the signal intensity and the resolution are maintained.

ADVANTAGEOUS EFFECTS OF INVENTION

According to the embodiment, it is possible to realize imaging with maintained signal intensity and resolution.

Drawings

Fig. 1 is a schematic configuration diagram of a confocal microscope a according to an embodiment.

Fig. 2 is a plan view showing a refraction state of the fluorescent light in the dichroic mirrors 9a to 9 c.

Fig. 3 is a cross-sectional view showing a mounting structure of the confocal microscope unit 1 of fig. 1 to the microscope 50.

Fig. 4 is a graph showing the wavelength distribution characteristics of the excitation light and the fluorescence processed by the 1 st to 4 th subunits 6a to 6d of fig. 1.

Fig. 5 is a diagram showing an image of light guide of excitation light or fluorescence in the confocal microscope unit 1.

Fig. 6 is a diagram showing an image of light guide of excitation light or fluorescence in the confocal microscope unit 1.

Fig. 7 is a diagram showing a configuration of a jig used for the oscillation adjustment of the confocal microscope unit 1.

Detailed Description

Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. In the description, the same elements or elements having the same function are denoted by the same reference numerals, and redundant description thereof will be omitted.

Fig. 1 is a schematic configuration diagram of a confocal microscope a as a type of scanning microscope according to an embodiment. The confocal microscope a shown in fig. 1 is configured to acquire a possible image constructed by constructing an optical tomographic image of a sample M to be observed, and the confocal microscope unit 1, which is a scanning microscope unit according to the embodiment, is configured to be connected to a connection port P1 for connecting an external unit of the microscope 50. The confocal microscope unit 1 is a device that irradiates an excitation light to a sample M placed on a stage of a microscope 50 and the like via a microscope optical system such as an imaging lens 51 and an objective lens 52 in the microscope 50, receives (detects) fluorescence generated from the sample M by the excitation light via the microscope optical system of the microscope 50, generates an optical tomographic image, and outputs the optical tomographic image: .

Specifically, the confocal microscope unit 1 includes a main housing 2, a lens barrel (housing) 3 constituting a part of the main housing 2 and detachably connected to a connection port P1 of the microscope 50, a scanning mirror 4 fixed in the main housing 2, a fixed mirror 5, 1 st to 4 th sub-units 6a to 6d, and a scanning lens 7 fixed in the lens barrel 3. Hereinafter, each constituent element of the confocal microscope unit 1 will be described in detail.

The scanning lens 7 in the lens barrel 3 is an optical element for converging the excitation light (irradiation light) to the 1 st image plane of the microscope optical system of the microscope 50 while relaying the reflection surface of the scanning mirror 4 to the pupil position of the objective lens 52. The scanning lens 7 guides the excitation light (irradiation light) scanned by the scanning mirror 4 to the microscope optical system to irradiate the sample M with the excitation light, and guides fluorescence (observation light) generated from the sample M according to the irradiation light to the scanning mirror 4. Specifically, the scanning lens 7 is configured to form an image of the pupil of the objective lens 52 on the scanning mirror 4, and guide the fluorescence formed by the objective lens 52 and the imaging lens 51 of the microscope 50 to the scanning mirror 4.

The scanning mirror 4 in the main casing 2 is an optical scanning element such as a Micro Electro Mechanical System (MEMS) mirror in which a reflection plate is formed to be tiltable in two axes, for example. The scanning mirror 4 has a function of scanning the excitation light (irradiation light) output from the 1 st to 4 th subunits 6a to 6d on the sample M by continuously changing the reflection angle, and guiding the fluorescence (observation light) generated by the excitation light to the 1 st to 4 th subunits 6a to 6 d.

The fixed mirror 5 is a light reflecting element fixed in the main casing 2, and reflects the excitation light output from the 1 st to 4 th sub-units 6a to 6d toward the scanning mirror 4, and reflects the fluorescence reflected by the scanning mirror 4 toward the 1 st to 4 th sub-units 6a to 6d coaxially with the excitation light.

The 1 st subunit 6a has a substrate 8a, a dichroic mirror (1 st beam splitter) 9a disposed on the substrate 8a, a light source 10a, a dichroic mirror 11a, a pinhole plate (1 st aperture member) 12a, and a photodetector (1 st photodetector) 13 a. The dichroic mirror 9a is a beam splitter having a property of reflecting the 1 st excitation light having the wavelength λ 1 irradiated by the 1 st subunit 6a and the 1 st fluorescence having the wavelength range Δ λ 1 generated from the sample M according to the reflected fluorescence, and transmitting light having a longer wavelength than the 1 st excitation light and the 1 st fluorescence, and fixed to the fixed mirror 5 in the reflection direction of the fluorescence. The dichroic mirror 11a is a beam splitter which is provided in the reflection direction of the 1 st fluorescence of the dichroic mirror 9a, and has properties of transmitting the 1 st fluorescence in the wavelength range Δ λ 1 and reflecting the 1 st excitation light having the wavelength λ 1 shorter than the wavelength range Δ λ 1. The light source 10a is a light emitting element (e.g., a laser diode) that outputs the 1 st excitation light (e.g., laser light) of the wavelength λ 1, and is arranged such that the 1 st excitation light is reflected by the dichroic mirror 11a toward the dichroic mirror 9a coaxially with the 1 st fluorescence. The pinhole plate 12a is an aperture member that is disposed so that the position of the pinhole coincides with the conjugate position of the light spot of the 1 st excitation light of the sample M and limits the light flux of the 1 st fluorescence, and constitutes a confocal optical system together with the light source 10a and the like. The pinhole plate 12a can adjust the diameter of the pinhole from the outside, and can change the resolution of the image detected by the photodetector 13a and the signal intensity of the image. The photodetector 13a is disposed so that its detection surface faces the pinhole plate 12a, and receives and detects the 1 st fluorescence that has passed through the pinhole plate 12 a. The photodetector 13a is a photomultiplier, a photodiode, an avalanche photodiode, an MPPC (Multi-Pixel Photon Counter), an HPD (Hybrid Photo Detector), an area image sensor, or the like.

The 2 nd to 4 th subunits 6b to 6d also have the same configuration as the 1 st subunit 6 a.

That is, the 2 nd subunit 6b has a substrate 8b, a dichroic mirror (2 nd beam splitter) 9b, a light source 10b, a dichroic mirror 11b, a pinhole plate (2 nd aperture member) 12b, and a photodetector (2 nd photodetector) 13 b. The dichroic mirror 9b has a property of reflecting the 2 nd excitation light of wavelength λ 2 (> λ 1) irradiated by the 2 nd subunit 6b and the 2 nd fluorescence of wavelength range Δ λ 2 generated from the sample M according to the reflected light, and transmitting light of a longer wavelength than the 2 nd excitation light and the 2 nd fluorescence. The dichroic mirror 11b has properties of transmitting the 2 nd fluorescence in the wavelength range Δ λ 2 and reflecting the 2 nd excitation light of the wavelength λ 2 shorter than the wavelength range Δ λ 2. The light source 10b is a light emitting element that outputs the 2 nd excitation light of the wavelength λ 2. The pinhole plate 12b is an aperture member that restricts the luminous flux of the 2 nd fluorescence, and the position of the pinhole coincides with the conjugate position of the spot of the 2 nd excitation light of the sample M. The photodetector 13b is disposed so that its detection surface faces the pinhole plate 12b, and receives and detects the 2 nd fluorescence that has passed through the pinhole plate 12 b. The photodetector 13b is a photomultiplier, a photodiode, an avalanche photodiode, an MPPC (Multi-Pixel Photon Counter), an HPD (Hybrid Photo Detector), an area image sensor, or the like.

The 3 rd subunit 6c has a substrate 8c, a dichroic mirror (3 rd beam splitter) 9c, a light source 10c, a dichroic mirror 11c, a pinhole plate (3 rd aperture member) 12c, and a photodetector (3 rd photodetector) 13 c. The dichroic mirror 9c has a property of reflecting the 3 rd excitation light of wavelength λ 3 (> λ 2) irradiated by the 3 rd subunit 6c and the 3 rd fluorescence of wavelength range Δ λ 3 generated from the sample M according to the reflected light, and transmitting light of a longer wavelength than the 3 rd excitation light and the 3 rd fluorescence. The dichroic mirror 11c has properties of transmitting the 3 rd fluorescence in the wavelength range Δ λ 3 and reflecting the 3 rd excitation light having the wavelength λ 3 shorter than the wavelength range Δ λ 3. The light source 10c is a light emitting element that outputs the 3 rd excitation light of the wavelength λ 3. The pinhole plate 12c is an aperture member that restricts the 3 rd fluorescence beam, with the pinhole position thereof being aligned with the conjugate position of the 3 rd excitation light spot of the sample M. The photodetector 13c is disposed so that its detection surface faces the pinhole plate 12c, and receives and detects the 3 rd fluorescence that has passed through the pinhole plate 12 c. The photodetector 13c is a photomultiplier, a photodiode, an avalanche photodiode, an MPPC (Multi-Pixel Photon Counter), an HPD (Hybrid Photo Detector), an area image sensor, or the like.

The 4 th subunit 6d has a substrate 8d, a total reflection mirror 9d, a light source 10d, a dichroic mirror 11d, a pinhole plate (4 th aperture member) 12d, and a photodetector (4 th photodetector) 13 d. The total reflection mirror 9c reflects the 4 th excitation light of the wavelength λ 4 (> λ 3) irradiated from the 4 th subunit 6d and the 4 th fluorescence of the wavelength range Δ λ 4 generated from the sample M in accordance therewith. The dichroic mirror 11d has properties of transmitting the 4 th fluorescence in the wavelength range Δ λ 4 and reflecting the 4 th excitation light having the wavelength λ 4 shorter than the wavelength range Δ λ 4. The light source 10d is a light emitting element that outputs the 4 th excitation light of the wavelength λ 4. The pinhole plate 12d is an aperture member that restricts the luminous flux of the 4 th fluorescence, and the position of the pinhole coincides with the conjugate position of the luminous spot of the 4 th excitation light of the sample M. The photodetector 13d is disposed so that its detection surface faces the pinhole plate 12d, and receives and detects the 4 th fluorescence that has passed through the pinhole plate 12 d. The photodetector 13d is a photomultiplier, a photodiode, an avalanche photodiode, an MPPC (Multi-Pixel Photon Counter), an HPD (Hybrid Photo Detector), an area image sensor, or the like.

The positional relationship between the 1 st to 4 th subunits 6a to 6d configured as described above will be described.

The 1 st to 4 th subunits 6a to 6d are fixed in the main casing 2 so as to be arranged in this order along the direction of separating from the fixed mirror 5 along the light guiding direction of the 1 st to 4 th fluorescent lights by the scanning mirror 4 and the fixed mirror 5, and so as to have the dichroic mirrors 9a to 9c and the total reflection mirror 9d positioned on the optical paths of the 1 st to 4 th fluorescent lights. Specifically, the 2 nd to 4 th subunits 6b to 6d are arranged to be shifted (shift) by a predetermined distance d in a direction perpendicular to the light guiding direction of the 2 nd to 4 th fluorescent lights with respect to the 1 st to 3 rd subunits 6a to 6c, respectively, with reference to the center positions of the dichroic mirrors 9a to 9c and the total reflection mirror 9 d.

The predetermined distance d is set to be substantially equal to a movement amount δ on the optical path of the fluorescent light transmitted by the dichroic mirrors 9a to 9c, the movement amount δ being a movement amount in a direction perpendicular to the optical path due to refraction of the fluorescent light in the dichroic mirrors 9a to 9 c. In the present embodiment, since the thicknesses of the mirror members constituting the dichroic mirrors 9a to 9c are set to be the same, the amounts of movement generated in the dichroic mirrors 9a to 9c are substantially the same, and accordingly, the moving distances d between the adjacent 2 sub-units among the 1 st to 4 th sub-units 6a to 6d are also set to be the same. The moving distance d is appropriately set according to the thickness and refractive index of the mirror members constituting the dichroic mirrors 9a to 9 c. Specifically, when the thickness of the mirror member is t, the refractive index is n, the incident angle of fluorescence incident on the mirror member is θ, and the refraction angle into the mirror member is Φ, the movement amount δ of fluorescence based on the mirror member is shown in fig. 2. In this case, the movement amount δ can be obtained as shown in the following equation (1), and therefore, the movement distance (predetermined distance) d may be set in accordance with the movement amount δ. In addition, the incident angle θ and the refraction angle Φ have a relationship of the following expression (2).

δ＝t·sin(θ-φ)/cosφ…(1)

φ＝arcsin(sinθ/n)…(2)

When the incident angle θ is 45 degrees, if the refractive index n of the mirror member is 1.5, d ═ δ is 0.33t, and if the refractive index n of the mirror member is 1.4, d ═ δ is 0.29 t.

Next, the mounting structure of the confocal microscope unit 1 to the microscope 50 will be described in detail. Fig. 3 is a sectional view showing a mounting structure of the confocal microscope unit 1 to the microscope 50.

As shown in fig. 3, a scanning lens 7 including a plurality of lenses is fixed inside the lens barrel 3, and a swing adjustment mechanism 23 in which a fitting portion 21 and a movable portion 22 are integrated is provided inside the front end of the lens barrel 3. The attachment 21 has a ring shape protruding from the distal end of the lens barrel 3, and has a structure (for example, a structure corresponding to a mount C (mount)) that can be attached to a camera connection port P1 of the microscope 50 on the distal end side. The movable portion 22 is formed continuously with the proximal end side of the metal fitting portion 21, has a substantially annular shape, and the outer surface thereof constitutes a sliding surface formed in a spherical shape. A spherical sliding surface 24 corresponding to the outer surface shape of the movable portion 22 is formed on the inner surface of the distal end of the lens barrel 3. Here, the outer surface of the movable portion 22 and the inner surface of the lens barrel 3 are formed in the shapes described below: in a state where the movable part 22 is fitted into the lens barrel 3 and the attachment part 21 is connected to the connection port P1 of the microscope 50, the center C1 of the spherical surface including these shapes is positioned on the imaging surface S1 of the microscope optical system of the microscope 50.

According to the mounting structure in which the swing adjustment mechanism 23 having the above-described structure is fitted to the distal end side of the lens barrel 3, the lens barrel 3 can be supported so that the angle of the lens barrel 3 with respect to the accessory part 21 can be changed by sliding the movable part 22 with respect to the sliding surface 24 of the lens barrel 3 in a state of being mounted on the microscope 50. At this time, since the outer surface of the movable portion 22 and the inner surface of the lens barrel 3 are formed in a spherical shape, the lens barrel 3 can be rotated with respect to the accessory portion 21, and the angle of the central axis of the lens barrel 3 with respect to the central axis of the accessory portion 21 can be two-dimensionally adjusted. That is, the swing adjustment mechanism 23 is configured to be able to change the angle of the lens barrel 3 with respect to the attachment part 21 so that the optical axis of the microscope optical system of the microscope 50 is parallel to the optical axis of the scanning lens 7.

According to the confocal microscope unit 1 described above, the excitation light (irradiation light) irradiated from the light sources 10a to 10d of the respective subunits 6a to 6d is scanned on the sample M via the scanning mirror 4, the scanning lens 7, and the external microscope 50, and the fluorescence (observation light) generated from the sample M based on the scanning light is detected by the photodetectors 13a to 13d of the respective subunits 6a to 6d via the external microscope 50, the scanning lens 7, and the scanning mirror 4. The lens barrel 3 to which the scanning lens 7 is fixed in the confocal microscope unit 1 is attached to the connection port P1 of the microscope 50 via the attachment part 21, and the angle of the lens barrel 3 with respect to the attachment part 21 can be changed. With such a configuration, the optical axis of the scanning lens 7 can be aligned in the direction of the optical axis of the microscope optical system of the microscope 50 corresponding to the microscope 50 to be mounted. As a result, it is possible to realize an image in which the signal intensity and the resolution are maintained.

Fig. 4 is a graph showing the wavelength distribution characteristics of the excitation light and the fluorescence processed by the 1 st to 4 th subunits 6a to 6 d. According to the wavelength lambda irradiated from the 1 st subunit 6a₁Wavelength range of fluorescence generated by the excitation light of (2)₁Usually to be at a wavelength λ₁And wavelength ratio wavelength λ₁A long range. In contrast, the wavelength λ of the excitation light emitted from the 2 nd subunit 6b₂And the wavelength range DeltaLambda of the fluorescence generated therefrom₂Becomes a wavelength ratio wavelength lambda₁Sum wavelength range Δ λ₁A long range. Here, the boundary wavelength λ at which the light of the dichroic mirror 9a of the 1 st subunit 6a is divided_d1Set as wavelength ratio wavelength lambda₁Sum wavelength range Δ λ₁Long and wavelength specific wavelength lambda₂Sum wavelength range Δ λ₂Short values. This enables the use of the wavelength λ of the 1 st subunit 6a₁Sum wavelength range Δ λ₁Can be measured using the 2 nd subunit 6b of the same device₂Sum wavelength range Δ λ₂Confocal measurement of the range of (1). Similarly, the boundary wavelength λ at which the light of the dichroic mirror 9b of the 2 nd subunit 6b is divided_d2Set as wavelength ratio wavelength lambda₂Sum wavelength range Δ λ₂Long and wavelength specific to wavelength lambda₃Sum wavelength range Δ λ₃Short value, boundary wavelength λ that divides light of dichroic mirror 9c of subunit 3c_d3Set as wavelength ratio wavelength lambda₃Sum wavelength range Δ λ₃Long and wavelength specific to wavelength lambda₄Sum wavelength range Δ λ₄Short values. This enables the 3 rd subunit 6c to use the same apparatus at the wavelength λ₃Sum wavelength range Δ λ₃Can be measured using the 4 th subunit 6d of the same device₄Sum wavelength range Δ λ₄Confocal measurement of the range of (1).

Here, the movable portion 22 of the rocking adjustment mechanism 23 is configured to be able to change the angle of the lens barrel 3 so that the optical axis of the scanning lens 7 is parallel to the optical axis of the microscope optical system of the microscope 50. In this case, the direction of the optical axis of the scanning lens 7 can be aligned with the optical axis of the microscope optical system, and the pupil positions of the scanning mirror 4 and the objective lens 52 can be arranged at conjugate positions, so that the NA of the objective lens 52 can be used to the maximum, and the signal intensity and the resolution of the signals detected by the photodetectors 13a to 13d can be reliably improved.

In particular, the movable portion 22 is configured to be rotatable with respect to the attachment portion 21. With this configuration, the direction of the scanning lens 7 can be two-dimensionally aligned with the optical axis of the microscope optical system, and the signal intensity and resolution of the signals detected by the photodetectors 13a to 13d can be reliably improved. The rotation center C1 of the movable unit 22 is included on the image forming surface S1 of the microscope optical system. In this way, the field of view can be adjusted without affecting the field of view, and the field of view adjustment and the angle adjustment do not need to be repeated alternately.

Fig. 5 shows an image of guided light of excitation light or fluorescence in the case where the direction of the optical axis of the scanning lens 7 in the confocal microscope unit 1 does not coincide with the direction of the optical axis of the microscope optical system, and fig. 6 shows an image of guided light of excitation light or fluorescence in the case where the direction of the optical axis of the scanning lens 7 in the confocal microscope unit 1 coincides with the direction of the optical axis of the microscope optical system.

In the confocal microscope unit 1, the microscope optics and the scanning lens 7 are adjusted in advance in such a way that the objective pupil of the microscope 50 is imaged on the scanning mirror 4. Thus, the change of the angle by the driving of the scanning mirror 4 is equivalent to swinging the angle of the beam of the excitation light or the fluorescence at the pupil of the objective lens 52, and the performance of the objective lens 52 can be sufficiently exhibited while reducing the loss of the beam. Here, the size of the objective lens pupil differs depending on the objective lens 52, and the beam diameter of the excitation light or the fluorescence needs to be equal to or larger than the pupil size, but in general, the scanning mirror 4 formed of a MEMS mirror or the like has a small diameter and a large tilt angle, and therefore, when the objective lens 52 of low magnification and low NA having a large pupil is used, the objective lens pupil may not converge on the scanning mirror 4.

In the microscope 50, the optical axis of the microscope may be inclined with respect to the connection port P1 (fig. 1) due to an assembly error. This tilt is a problem when imaging and detecting observation light, and is a problem particularly when used as a scanning microscope. As a result, as shown in fig. 4, when the confocal microscope unit 1 is mounted on the microscope 50, the optical axis a2 of the scanning lens 7 may be inclined at an angle θ (> 0) of about the optical axis a1 of the microscope optical system. Such a situation may cause a situation in which, when the scanning mirror 4 is driven to change the imaging position of the excitation light and the sample M is irradiated with the light rays B1, B2, the positions of the light rays B1, B2 may be deviated from the aperture of the objective lens, and the excitation light may be partially reflected. As a result, the amount of signals and the resolution of the images generated by the subunits 6a to 6d are reduced.

On the other hand, as shown in fig. 6, when the confocal microscope unit 1 is mounted on the microscope 50, the direction of the optical axis a2 of the scanning lens 7 is adjusted to be aligned with respect to the optical axis a1 of the microscope optical system by using the oscillation adjusting mechanism 23, so that the positions of the light beams B1 and B2 can be matched with the aperture position of the objective lens when the imaging position of the excitation light is changed and the sample M is irradiated with the light beams B1 and B2. As a result, the excitation light can be prevented from being reflected, and the signal amount and resolution of the images generated using the subunits 6a to 6d can be improved.

The swing adjustment using the swing adjustment mechanism 23 of the present embodiment can be realized using a jig as shown in fig. 7. Fig. 7 is a diagram showing a configuration of a jig used for the oscillation adjustment of the confocal microscope unit 1. As the jig, a jig including a disk-shaped target member 31 and a mounting portion 32, the mounting portion 32 being used to mount the target member 31 to the microscope 50 instead of the objective lens 52, can be employed. The center of the surface of the target member 31 is provided with a circular mark 33, and when the microscope 50 is attached by the attachment frame portion 32, the position of the mark 33 matches the pupil position of the objective lens 52.

With the jig having such a configuration, the excitation light is output from the confocal microscope unit 1, and the light spot SP1 of the excitation light irradiated to the target member 31 is observed with an external camera or the like. Then, by performing the oscillation adjustment so that the spot SP1 of the excitation light coincides with the mark 33 on the target member 31, it is possible to perform, for example, adjustment for aligning the direction of the optical axis of the scanning lens 7 with respect to the optical axis of the microscope optical system.

Furthermore, the scanning mirror 4 may also be a MEMS mirror. In this case, the device can be easily reduced in size.

Each of the subunits 6a to 6d includes a pinhole plate 12a to 12d that restricts the beam of observation light returned via the scanning mirror 4, and the photodetectors 13a to 13d are configured to detect the observation light that has passed through the pinhole plates 12a to 12 d. In this case, it is possible to realize an image in which the signal intensity and the resolution are maintained in the confocal observation.

While various embodiments of the present invention have been described above, the present invention is not limited to the above embodiments, and can be modified within the scope not changing the gist of the claims, or can be applied to other embodiments.

The swing adjustment mechanism 23 of the above embodiment is not limited to the structure of the movable portion 22 having a spherical sliding surface, and the movable portion 22 may be formed of a rubber member or a bellows-shaped member.

In the above embodiment, the excitation light is irradiated to each of the subunits 6a to 6d, and the fluorescence generated by the excitation light can be detected, but the fluorescence detection unit may be configured to detect the reflected light generated in the sample M by irradiation of the irradiation light.

The above-described embodiment is not limited to the use as a confocal microscope, and may be used in a general fluorescence microscope, a reflection microscope, or the like as long as it is a scanning microscope using a scanning mirror.

The above embodiment is not limited to the application to a scanning microscope using a scanning mirror, and is also effective when the optical unit having a lens is attached to the connection port P1 of an optical microscope, and the housing of the optical unit is tilted so that the optical axis of the microscope optical system of the microscope 50 is parallel to the optical axis of the lens of the optical unit. In this case, the optical axes of the optical microscope and the optical unit are easily adjusted.

In the above-described embodiments, the pinhole plate is used as the aperture member to constitute the confocal optical system, but the aperture member may be any optical element that restricts a light flux, and may be, for example, a color aperture or an optical fiber core. When a fiber output type light source is used, the position of the end face of the optical fiber core may be set as the aperture position (the position where the light flux is restricted).

In the above embodiment, a laser light source such as a solid laser or a diode laser can be used. In this case, the position of the beam waist of the laser light source may be set as a diaphragm position (a position where the beam is limited), and the light source itself may function as a diaphragm member.

In the above embodiment, the movable portion may be a member capable of changing the angle of the housing so that the optical axis of the scanning lens is parallel to the optical axis of the microscope optical system. In this case, by aligning the direction of the scanning lens with the optical axis of the microscope optical system, the signal intensity and resolution of the signal detected by the photodetector can be reliably improved.

The movable portion may be configured to be rotatable with respect to the attachment portion. With this configuration, the direction of the scanning lens and the optical axis of the microscope optical system can be aligned two-dimensionally, and the signal intensity and resolution of the signal detected by the photodetector can be reliably improved.

The rotation center of the movable unit may be included in the image forming surface of the microscope optical system. In this case, the effect of increasing the amount of signals detected by the photodetector and improving the resolution can be obtained without affecting the field of view.

Further, the scanning mirror may also be a MEMS mirror. In this case, the device can be easily reduced in size.

The photodetector may detect fluorescence generated by irradiation of the irradiation light as the observation light, or may detect reflected light generated by irradiation of the irradiation light as the observation light. With this configuration, it is possible to realize imaging with maintained signal intensity and resolution in imaging with various types of observation light.

Further, the present invention may further include: and an aperture member that restricts a light flux of the observation light returned via the scanning mirror, wherein the light detector detects the observation light having passed through the aperture member. In this case, an optical tomographic image can be acquired by confocal observation. Further, the aperture member may be a pinhole plate.

Industrial applicability of the invention

In the embodiment, the scanning microscope unit constituting the scanning microscope is used for a purpose of use, and an image with maintained signal intensity and resolution can be obtained.

Description of the symbols

Optical axes of a1 and a2 …, a rotation center of C1 …, an M … sample, a P1 … connection port, an S1 … imaging plane, a d … specified distance, 10a to 10d … light sources, 12a to 12d … pinhole plates (diaphragm members), 13a to 13d … photodetectors, 6a to 6b … 1 st to 4 th subunits, 9a to 9C … dichroic mirrors, 1 … confocal microscope unit, 2 … main housing, 3 … lens barrel (housing), 4 … scanning mirror, 7 … scanning lens, 21 … accessory part, 22 … movable part, 23 … swing adjustment mechanism, 24 … sliding surface, 50 … microscope, 52 … objective lens.