CN210954468U

CN210954468U - Microscope device

Info

Publication number: CN210954468U
Application number: CN201890000408.0U
Authority: CN
Inventors: 大坪诚
Original assignee: Asukanet Co Ltd
Current assignee: Asukanet Co Ltd
Priority date: 2017-04-11
Filing date: 2018-04-04
Publication date: 2020-07-07
Anticipated expiration: 2028-04-04
Also published as: JP6632762B2; CN212749367U; JP2020034956A; WO2018190221A1; JPWO2018190221A1

Abstract

A microscope device (10) is provided with: a support unit 11a that holds a sample (11) to be observed; an illumination unit (17) that irradiates the sample (11) with light from a specific direction; and a microscope (13) disposed facing an observation target to be magnified and observed, wherein a three-dimensional image imaging unit (12) is disposed between the sample (11) and an objective lens (15) of the microscope (13), the sample (11) is disposed on one side of the three-dimensional image imaging unit (12), a real image (14) of the sample (11) is formed on the other side of the three-dimensional image imaging unit (12) by the three-dimensional image imaging unit (12), and the real image (14) is used as the observation target of the microscope (13).

Description

Microscope device

Technical Field

The present invention relates to a microscope device capable of observing a sample as an observation target while keeping the sample (in a planar or three-dimensional state).

Background

Conventionally, a sample is observed using a microscope (optical microscope) by placing a sample (i.e., a slice) that is attached to a slide glass and sealed with a cover glass on a stage (see, for example, patent document 1).

Documents of the prior art

Patent document

Patent document 1: japanese patent laid-open publication No. 2007-17806

Patent document 2: japanese patent No. 5437436

Patent document 3: WO2006/001158 publication

Patent document 4: japanese patent No. 6203989

SUMMERY OF THE UTILITY MODEL

Problem to be solved by utility model

However, in patent document 1, since the specimen is observed in a state of being sandwiched between the slide glass and the cover glass, the specimen becomes planar, and the specimen cannot be observed in a state of being kept as it is (original state). In addition, although there are some microscopes for observing a three-dimensional image of a sample, such as a confocal laser scanning microscope and a multiphoton laser excitation laser scanning microscope, in this case, the device configuration of the microscope becomes complicated, the cost is high, the economical efficiency is poor, and the operability is also problematic.

Further, when a specimen is observed using a slice, the objective lens may collide with the slice and damage the objective lens when focus adjustment is performed.

Patent documents 2 to 4 propose a display device for stereoscopic images and a method for manufacturing the same, but do not enlarge an image of a sample.

The present invention has been made in view of such circumstances, and an object thereof is to provide a microscope device that can realize, with a simple device configuration, 1) observation of a transparent or translucent sample at a desired arbitrary depth position without damaging an objective lens, for example, while keeping the sample intact; and 2) depending on the case, even if the specimen is opaque, the specimen can be observed by enlarging the specimen without bringing the specimen or the slice into contact with the objective lens.

Means for solving the problems

To the purpose the utility model discloses a microscope device has: a support unit that holds a sample to be observed; an illumination unit that irradiates the sample with light from a specific direction; and a microscope disposed toward an observation target to be magnified and observed, wherein a three-dimensional image forming unit is disposed between the sample and an objective lens of the microscope, the sample is disposed on one side of the three-dimensional image forming unit, a real image of the sample is formed in a space on the other side of the three-dimensional image forming unit by the three-dimensional image forming unit, and the real image is set as the observation target of the microscope.

In the microscope device of the present invention, the three-dimensional image forming means is preferably formed by arranging a 1 st light control panel and a 2 nd light control panel each having a plurality of strip-shaped light reflecting portions arranged in parallel, in a manner such that the light reflecting portions intersect (preferably orthogonally intersect) in a plan view (see, for example, patent documents 2 and 4).

In the microscope device of the present invention, it is preferable that the supporting means is a transparent container filled with a liquid having translucency, the transparent or translucent sample is placed in the liquid in the container, and the magnified cross section of the sample can be observed by adjusting the focus of the microscope.

Here, the real image may be formed by two of the stereoscopic image forming units optically arranged in series.

In the microscope device of the present invention, the support unit may be a transparent plastic body, and the sample may be contained or sealed in the plastic body.

In the microscope device of the present invention, the illumination unit irradiates light from a specific direction onto a sample to be observed which is held by the support unit and arranged on one side of the three-dimensional imaging unit, thereby forming a real image of the sample on the other side of the three-dimensional imaging unit and observing the real image with the microscope.

In the microscope device according to the present invention, it is preferable that the three-dimensional image forming means is formed by arranging a 1 st light control panel and a 2 nd light control panel each having a plurality of strip-shaped light reflection portions arranged in parallel in a state where the light reflection portions intersect (preferably intersect orthogonally) in a plan view, and the light from the sample reflected by the light reflection portion of the 1 st light control panel is reflected again by the light reflection portion of the 2 nd light control panel to form a real image of the sample.

Effect of the utility model

In the microscope device of the present invention, since the three-dimensional image forming means is disposed between the sample and the objective lens of the microscope and the sample is disposed on one side of the three-dimensional image forming means, the real image of the sample is formed on the other side by the three-dimensional image forming means, and the real image can be magnified and observed by the microscope. Thus, for example, a transparent or translucent sample can be imaged in a space in a state of being held as it is by a simple apparatus configuration, and the image formed in the space can be observed by the objective lens, and therefore, an observation target can be observed at an arbitrary depth position without damaging the objective lens. In addition, even an opaque sample can be observed by enlarging the sample without bringing the sample or the slice into contact with the objective lens, depending on the case.

Drawings

Fig. 1 is an explanatory view of a microscope device according to an embodiment of the present invention.

Fig. 2 is an explanatory view showing a state of formation of a real image of a sample formed by the stereoscopic image forming unit of the microscope apparatus.

Fig. 3 (a) and (B) are a front sectional view and a side sectional view of the stereoscopic imaging unit of the microscope device, respectively.

Fig. 4 (a) to (C) are explanatory views of the sample container used in the microscope apparatus.

Detailed Description

Next, specific embodiments of the present invention will be described with reference to the drawings for understanding the present invention.

As shown in fig. 1 and 2, a microscope device 10 according to an embodiment of the present invention is a device including: a stage (an example of a support unit) 11a that holds a sample 11; a stereoscopic image imaging unit 12; and a microscope 13 for observing a real image (an example of an observation target) 14 of the sample 11 formed by the stereoscopic image forming unit 12 by enlarging the microscope 13 disposed toward the real image 14, whereby the sample 11 can be observed in a state of being kept as it is (in a stereoscopic state) without making the sample 11 planar. The details will be described below.

The microscope 13 is a conventionally known optical microscope, and includes, for example, an inverted microscope, a measuring microscope, a dissecting microscope, a phase-difference microscope, a differential interference microscope, a polarization microscope, a fluorescence microscope, and the like.

The microscope 13 includes an objective lens 15 and an eyepiece lens 16, and forms an image of a real image 14 (also referred to as an inverted real image, an intermediate image, or a primary magnified image) through the objective lens 15, and guides the image to the eyepiece lens 16 to form an erect virtual image. The microscope 13 is a compound microscope having an objective lens 15 and an eyepiece 16, but may be a single microscope for directly observing an inverted real image magnified by the objective lens, and may be a television observation microscope (i.e., a microscope without an eyepiece) for directly capturing an inverted real image by a CCD camera or the like.

The microscope apparatus 10 is provided with an illumination unit 17, and as shown in fig. 1, is configured to irradiate the sample 11 with light from the left side (specific direction) in the horizontal direction. As the illumination unit 17, for example, a light source such as an LED lamp or a halogen lamp can be used. In fig. 1, reference numeral 18 denotes a light reflecting mirror provided on the microscope 13, and does not have to be provided.

The sample 11 to be observed by the microscope device 10 is a transparent or translucent sample that can be observed by an optical microscope and can be visually confirmed from the outside. Specifically, microorganisms such as paramecium or daphnia, human cells, and fungi such as escherichia coli are present. The sample may be opaque depending on the case.

A stereoscopic image imaging unit 12 is disposed between the objective lens 15 of the microscope 13 and the sample 11.

As shown in fig. 3 (a) and (B), the three-dimensional image forming unit 12 is formed by overlapping and arranging a 1 st light control panel (parallel light reflection panel) 20 and a 2 nd light control panel (parallel light reflection panel) 20a such that the vertical light reflection surface 19 of the 1 st light control panel 20 and the vertical light reflection surface 19 of the 2 nd light control panel 20a are orthogonal (an example of intersection, for example, may intersect in a range of 88 to 92 degrees) in a plan view (for example, see patent document 4 and PCT/JP2017/12622), wherein each of the 1 st light control panel (parallel light reflection panel) 20 and the 2 nd light control panel (parallel light reflection panel) 20a has a plurality of strip-shaped vertical light reflection surfaces (an example of light reflection portions) 19 arranged in parallel (standing up). Since the 1 st and 2 nd

light control panels

20 and 20a have the same configuration, the same reference numerals are given to the components.

The 1 st light control panel 20 (the same applies to the 2 nd light control panel 20 a) has a groove 24 having a triangular cross section including a vertical surface 22 and an inclined surface 23 and a convex strip 25 having a triangular cross section formed between the grooves 24 on one side (an upper portion for the 1 st light control panel 20 and a lower portion for the 2 nd light control panel 20 a) of the transparent flat plate 21. The grooves 24 and the ridges 25 of the 1 st and 2 nd light control panels 20 are provided in parallel at a fixed pitch.

A metal reflective film (metal plating film) 22a is formed on the vertical surface 22 of the groove 24 by mirror surface processing, and one surface of the metal reflective film 22a becomes the vertical light reflective surface (mirror surface) 19.

On the other hand, the inclined surface 23 is preferably a non-light reflecting surface, that is, a light transmitting surface.

The groove 24 is filled with a transparent resin 26, and the filling surfaces 27 are parallel to the surfaces 28 of the 1 st and 2 nd

light control panels

20 and 20a (e.g., the transparent flat plate 21), respectively.

The 1 st and 2 nd

light control panels

20 and 20a are arranged in contact with or in close proximity to each other in a state where the vertical light reflecting surfaces 19 thereof are perpendicular to each other in a plan view. The 1 st and 2 nd

light control panels

20 and 20a may be joined and integrated by a transparent adhesive (resin), for example.

The transparent resin constituting the shape (transparent flat plate 21 and convex strips 25) of the 1 st and 2 nd

light control panels

20 and 20a and the transparent resin 26 filled in the grooves 24 are preferably the same resin, but may be different kinds of transparent resins, and when different kinds of transparent resins are used, the refractive index is preferably the same or similar, that is, a transparent resin having the same or substantially the same refractive index (for example, in the range of ± 20%, that is, in the range of (0.8 to 1.2) η) as the refractive index (η) of the transparent resin constituting the shape of the 1 st and 2 nd

light control panels

20 and 20a is used as the transparent resin 26 filled in the grooves 24.

Thus, in fig. 3 a and B, light L1 and L2 from the sample 11 obliquely incident from one side (lower left side) of the three-dimensional image forming unit 12 is reflected by the P1 and P2 of the vertical light reflecting surface 19 of the lower 1 st light controlling panel 20, and then reflected again by the Q1 and Q2 of the vertical light reflecting surface 19 of the upper 2 nd light controlling panel 20a, whereby the real image 14 (three-dimensional image) can be formed on the other side (upper right side) of the three-dimensional image forming unit 12.

Here, the rear surface side (left side in fig. 3) of the metal reflection film 22a formed on the vertical surface 22 by the mirror surface processing is used as the vertical light reflection surface 19 of the 1 st and 2 nd

light control panels

20 and 20a, but the front surface side (right side in fig. 3) of the metal reflection film 22a may be used as the vertical light reflection surface 19.

The 1 st light control panel 20 (and likewise the 2 nd light control panel 20 a) can be manufactured by the following method: after a molding base material made of a transparent resin, in which grooves 24 each having a triangular cross section and a vertical surface 22 and an inclined surface 23 are formed in parallel on one side of a transparent plate 21, and ridges 25 each having a triangular cross section and formed by adjacent grooves 24, is manufactured, a metal reflective film 22a made of, for example, aluminum, silver, nickel, or the like is formed on the vertical surfaces 22 of the ridges 25 (grooves 24).

The molding base material may be manufactured by press molding, injection molding, or roll molding.

The metal reflective film 22a may be formed by sputtering, metal vapor deposition, or metal fine particle spray coating on the vertical surfaces 22 of the ridges 25.

The pitch of the metal reflective film 22a (vertical light reflecting surface 19) is not particularly limited, but is preferably 1 to 20 μm (more preferably 5 to 10 μm) in consideration of the size of the sample 11.

The three-dimensional image forming means is not limited to the above configuration as long as it can form a real image of the sample.

In addition to the case where the vertical surface on which the light reflecting portion is formed as the convex strip having a triangular cross section as described above, for example, as described in japanese patent No. 4865088, the light reflecting portion may be formed on one surface of a stacked transparent plate having a rectangular cross section.

The stereoscopic image forming unit 12 is formed by manufacturing the 1 st and 2 nd

light control panels

20 and 20a separately and overlapping them, but may be formed by forming grooves and ridges on the front and back surfaces of a transparent flat plate and integrally molding the 1 st and 2 nd light control panels.

The observation of the sample 11 by the microscope apparatus 10 is performed as follows: as shown in fig. 1, a sample 11 is placed and held on a planar stage (an example of a support unit) 11a provided in a microscope apparatus 10, a three-dimensional image forming unit 12 is disposed between the sample 11 and an objective lens 15, the sample 11 is disposed on one side (lower left side in fig. 1) of the three-dimensional image forming unit 12, and the sample 11 is irradiated with light from a horizontal direction by an illumination unit 17. Thus, since the sample 11 is transparent or translucent, a real image 14 of the sample 11 is formed by the stereoscopic imaging unit 12 at a position symmetrical to the sample 11 (the other side (upper right side in fig. 1) of the stereoscopic imaging unit 12) with the stereoscopic imaging unit 12 as the center (with the stereoscopic imaging unit 12 interposed therebetween), and the real image 14 becomes an observation target to be observed by the microscope 13 under magnification.

Here, the observation of the sample 11 may be performed in a state where the sample 11 is placed (loaded) in a petri dish (plate) as an example of a plastic body having translucency, or may be performed by placing a plastic body having translucency (transparent) in which the sample 11 is sealed on a stage 11a instead of the petri dish (in this case, the plastic body and the stage 11a serve as a support means).

As a method of sealing the sample 11 in a plastic body, there is a method of filling the sample 11 in a molten resin (resin filling) and curing the resin (a method of curing the resin by using heat or ultraviolet rays depending on the type of the resin).

As shown in fig. 4 (a) to (C), the observation of the sample 11 may be performed using containers 31 to 33 into which the sample 11 can be loaded as supporting means for holding the sample 11. The containers 31 to 33 are preferably transparent and made of a transparent plastic material, but may be made of another transparent material such as glass.

The containers 31 to 33 are filled with a liquid 34 having translucency. The liquid 34 may be, for example, water, physiological saline, or a culture solution (the same applies to the case of using a petri dish) depending on the type of the sample 11. The positioning of the sample 11 may be performed by freezing (becoming solid) the liquid 34 in a state where the sample 11 is present.

The container 31 shown in fig. 4 (a) has an opening 35 formed in the upper portion thereof in a horizontal state. The stereoscopic imaging unit 12 is attached and fixed to the upper portion of the container 31 in an inclined manner with the opening 35 of the container 31 opened. At this time, the microscope 13 is disposed so that the observation direction becomes a lateral direction (horizontal direction) toward the real image 14.

Thus, the real image 14 of the sample 11 (loaded in the container 31) in the liquid 34 filled in the container 31 can be formed on the side of the container 31 in a plan view at a position symmetrical to the sample 11 with the stereoscopic imaging unit 12 interposed therebetween.

The container 32 shown in fig. 4 (B) has an opening 36 formed in an inclined state at an upper portion thereof. The stereoscopic imaging unit 12 is attached and fixed to the upper end of the container 32 so as to close the opening 36. At this time, the microscope 13 is disposed so that the observation direction becomes a lateral direction (horizontal direction) toward the real image 14.

Thus, since air can be discharged from the container 32 by filling the liquid 34 into the container 32 sealed by the stereoscopic imaging unit 12, the real image 14 of the sample 11 in the liquid 34 can be formed on the side of the container 32 in a plan view at a position symmetrical to the sample 11 with the stereoscopic imaging unit 12 therebetween, without considering refraction between the liquid 34 and the air layer.

In the container 33 shown in fig. 4 (C), 2 stereoscopic image imaging units 12 are provided. The 2 stereoscopic imaging units 12 are optically arranged in series, and are arranged in a mountain shape with their tops abutting, and one inclined stereoscopic imaging unit 12 constitutes the bottom of the container 33. Therefore, the liquid 34 does not enter between the 2 stereoscopic imaging units 12 arranged in the mountain shape.

Although the upper end portions of the 2 stereoscopic imaging units 12 are in contact with each other here, the 2 stereoscopic imaging units 12 may be arranged in a separated state.

Thus, the real image 14a of the sample 11 formed by one of the three-dimensional image forming units 12 at a position symmetrical to the sample 11 in the liquid 34 about the one of the three-dimensional image forming units 12 is formed again by the other one of the three-dimensional image forming units 12 as a real image (an example of an observation target) 37 at a position symmetrical to the real image 14a about the one of the three-dimensional image forming units 12. At this time, the microscope 13 is disposed so that the observation direction becomes vertical (vertical direction) toward the real image 37.

Therefore, the real image 37 of the sample 11 to be observed is an image formed via the 2 stereoscopic image forming units 12 optically arranged in series, and therefore the irregularities are in a normal state (the same state as the sample 11).

Next, a sample observation method using the microscope device 10 according to one embodiment of the present invention will be described with reference to fig. 1 and 2.

First, as shown in fig. 1, a specimen 11 to be observed is arranged on the stage 11a of the microscope apparatus 10 on the side of the stereoscopic image imaging unit 12. Further, the stereoscopic imaging unit 12 is disposed on the stage 11a, and the sample 11 is disposed in the petri dish as described above, but containers 31 to 33 shown in fig. 4 (a) to (C) may be used instead of the petri dish. Then, the sample 11 is irradiated with light from a specific direction by the illumination unit 17.

As a result, the real image 14 of the sample 11 shown in fig. 2 (or the real image 37 shown in fig. 4C) is formed on the other side of the stereoscopic image imaging unit 12.

The cross-sectional image of the real image 14 of the sample 11 thus formed is enlarged by the objective lens 15 to form an enlarged cross-sectional image, and the enlarged cross-sectional image is enlarged by the eyepiece lens 16 to form a further enlarged cross-sectional image, which can be visually observed.

Specifically, when the sectional image of the real image 14 of the sample 11 is mn, an inverted real image 1 time (enlarged sectional image) m 'n' is formed by the objective lens 15. Further, by disposing the eyepiece 16 so that the 1 st order image m 'n' is positioned closer to the eyepiece 16 than the front focal point of the eyepiece 16, the erect virtual image MN is formed which is further magnified, and the magnified image can be observed by the eyes (pupils).

At this time, although the magnified image (magnified cross section) of the cross section at each position of the real image 14 can be observed by moving the objective lens 15 closer to or farther from the real image 14 (performing focus adjustment of the microscope 13), the objective lens 15 may enter the real image 14 depending on the position of the real image 14 to be observed. However, the real image 14 is not the sample 11 itself, but an image of the sample 11, and therefore the objective lens 15 is not damaged.

Therefore, according to the microscope device 10 and the sample observation method using the microscope device 10 of the present invention, it is possible to observe a transparent or translucent sample at an arbitrary depth position with a simple device configuration, while maintaining the original state, without damaging the objective lens. In addition, even an opaque sample, depending on the case, can be observed under magnification without bringing the sample or the slice into contact with the objective lens.

The present invention has been described above with reference to the embodiments, but the present invention is not limited to the configurations described in any of the above embodiments, and includes other embodiments and modifications that can be considered within the scope of the items described in the claims. For example, the scope of the present invention also includes a case where a part or all of the above-described embodiments or modifications are combined to constitute the microscope device of the present invention and the sample observation method using the microscope device.

For example, although the microscope provided in the microscope apparatus of the present invention is configured on the assumption that the stereoscopic imaging unit is disposed, the scope of the present invention is also included in the case where the stereoscopic imaging unit is provided in a conventionally used microscope apparatus (a microscope apparatus not on the assumption that the stereoscopic imaging unit is disposed).

Industrial applicability

The microscope device and the sample observation method using the same of the present invention can form an image of a sample in a space with a simple device configuration in a state where the sample is held, and can observe the image (real image) formed in the space by magnifying the image with an objective lens. This enables observation of an observation target at an arbitrary depth position, and thus can contribute to, for example, research of organisms (animals, plants, fungi, and bacteria), development of medicine, development of drugs, and the like.

Description of the reference symbols

10: a microscope device; 11: a sample; 11 a: a stage (support unit); 12: a stereoscopic image imaging unit; 13: a microscope; 14: real images (observation objects); 14 a: real images; 15: an objective lens; 16: an eyepiece; 17: a lighting unit; 18: a mirror; 19: a vertical light reflecting surface (light reflecting section); 20: a 1 st light control panel; 20 a: a 2 nd light control panel; 21: a transparent flat plate; 22: a vertical plane; 22 a: a metal reflective film; 23: an inclined surface; 24: a groove; 25: a convex strip; 26: a transparent resin; 27: a filling surface; 28: a surface; 31-33: a container; 34: a liquid; 35. 36: an opening part; 37: real image (observation target).

Claims

1. A microscope device, having: a support unit that holds a sample to be observed; an illumination unit that irradiates the sample with light from a specific direction; and a microscope arranged toward an observation object to be observed under magnification,

a three-dimensional image forming unit is disposed between the sample and an objective lens of the microscope, the sample is disposed on one side of the three-dimensional image forming unit, a real image of the sample is formed in a space on the other side of the three-dimensional image forming unit by the three-dimensional image forming unit, and the real image is set as the observation target of the microscope.

2. The microscope device according to claim 1,

the three-dimensional image forming unit is formed by overlapping and arranging a 1 st light control panel and a 2 nd light control panel, each of which has a plurality of strip-shaped light reflecting portions arranged in parallel, in a state where the light reflecting portions intersect when viewed from above.

3. The microscope device according to claim 1 or 2,

the supporting means is a transparent container filled with a liquid having translucency, the transparent or semitransparent sample is placed in the liquid in the container, and the magnified cross section of the sample can be observed by adjusting the focus of the microscope.