JPH10325798A - Microscope apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microscope apparatus in which both a visible-light image by ordinary observation light and a fluorescence image emitted from a body tissue by an indosyanine green derivative labeled antibody are observed simultaneously. SOLUTION: A sample 2 which is stained with an indocyanine green derivative labeled antibody is irradiated with beams of illumination light from lamps 41, 51 by which light in a first wavelength band containing at least a part of the excition wavelength of a fluorescent substance and light in a second wavelength band containing at least a part of visible light are radiated. Thereby, a light image which contains reflected light and infrared light is emitted from the sample 2. The light image is separated into a fluorescence (infrared light) component and a visible-light (reflected light) component by using a second dischroic mirror 70. A fluorescence image and a visible-light image which are imaged and obtained by a CCD 71 and a CCD 72 corresponding to the light image for the respective components are observed and diagnosed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a microscope apparatus for observing an indocyanine green derivative-labeled antibody.

[0002]

2. Description of the Related Art In recent years, the spread of endoscopes has facilitated the detection of gastric cancer and colorectal cancer. However, regarding the diagnosis of small cancer, it is necessary to judge whether or not cancer is present only from an endoscopic image. It was very difficult. For this reason, when diagnosing microcancer, a part of the suspicious body tissue is excised through a treatment instrument channel provided in the endoscope, and the excised tissue is observed under a microscope to determine whether there is cancer. And the extent of infiltration was determined.

[0003] When diagnosing cancer under a microscope, a method for identifying a diseased tissue from the shape of cells or the like, or a method for staining a section of a body tissue extracted from the body by reacting it with an antibody that specifically binds to the cancer is used. Is used. In this staining method, for example, a diagnostic marker is allowed to act on a section of a body tissue extracted from the body, and a color is developed by an avidin / bition complex method (ABC method). Diamoben
Color development is observed due to the deposition of zidine).

Recently, indocyanine green (hereinafter referred to as ICG) which is excited by near-infrared rays and emits near-infrared fluorescence.
Has also been attempted to be used as a diagnostic marker by binding to an antibody. This indocyanine green derivative-labeled antibody has the international publication number WO96 /
As described in US Pat. No. 23,525, it has an affinity for a focus such as cancer, and is accumulated in a diseased tissue by reacting with a sample.

[0005]

However, in the case of making a diagnosis using the above-described DAB coloring, there is a problem that enzymatic reactions and the like take a long time in the pretreatment stage, so that the test cannot be performed quickly. Was. Further, there has not been proposed a microscope apparatus for observing an indocyanine green derivative-labeled antibody.

The present invention has been made in view of the above circumstances, and provides a microscope apparatus capable of simultaneously observing both a visible light image by ordinary observation light and a fluorescence image emitted from a body tissue by an indocyanine green derivative-labeled antibody. It is aimed at.

[0007]

A microscope apparatus according to the present invention comprises:
Light source means for irradiating a sample containing a fluorescent substance with light in a first wavelength band containing at least a part of the excitation wavelength of the fluorescent substance and light in a second wavelength band containing at least a part of visible light. Light for spatially separating light emitted from the sample into light in a third wavelength band including at least a part of the fluorescent wavelength of the fluorescent substance and light in a fourth wavelength band including at least a part of visible light A separating unit; a first imaging unit configured to image light including at least a part of the third wavelength band;
Second imaging means for imaging light containing at least a part of the wavelength band.

According to this configuration, the light of the third wavelength band including at least a part of the fluorescence wavelength emitted from the sample and the light of the fourth wavelength band including at least a part of the visible light are respectively transmitted to the first imaging unit. , The image obtained by the second imaging means and the image obtained by the light of the third wavelength band and the image obtained by the light of the fourth wavelength band can be simultaneously observed.

[0009]

Embodiments of the present invention will be described below with reference to the drawings. 1 to 6 relate to a first embodiment of the present invention, FIG. 1 is an explanatory diagram showing a schematic configuration of the entire microscope apparatus, FIG. 2 is a characteristic diagram for explaining excitation / fluorescence characteristics of an indocyanine green derivative-labeled antibody, FIG. 3 is a characteristic diagram showing the spectral transmission characteristics of the bandpass filter, FIG. 4 is a characteristic diagram showing the spectral transmission characteristics of the first dichroic mirror, and FIG.
Is a characteristic diagram showing a spectral transmission characteristic of the second dichroic mirror, and FIG. 6 is a characteristic diagram showing a spectral transmission characteristic of the excitation light cut filter.

As shown in FIG. 1, a microscope apparatus 1 according to this embodiment includes an XY stage (hereinafter abbreviated as a stage) 3 on which a sample 2 such as a section of a body tissue is placed, and this stage 3
A transmission illumination optical system 4 which is a light source means for irradiating illumination light for observation from the back side of the sample 2 arranged thereon, and illumination light for observation is illuminated from the front side of the sample 2 arranged on the stage 3. An epi-illumination optical system 5 as a light source means, and the sample 2
A visual observation system 6 for observing the optical image of the sample 2 with the naked eye, and an imaging observation system 7 for capturing an optical image of the sample 2 and displaying the optical image as an image on a monitor screen for observation. It is configured. The sample 2 was previously stained with an indocyanine green derivative-labeled antibody.

The excitation and fluorescence characteristics when the indocyanine green derivative-labeled antibody is bound to human IgG are as shown in FIG. 2, and the peak wavelength of the excitation is around 770 nm as shown by the broken line. The peak wavelength is around 810 nm as shown by the solid line.

The transmission illumination optical system 4 emits light of a first wavelength band including at least a part of the excitation wavelength of the fluorescent substance and light of a second wavelength band including at least a part of the visible light. Lamp 41, a first collector lens 42 for condensing illumination light from the first lamp 41,
A first band-pass filter 43 that transmits a desired wavelength band of the illumination light transmitted through the collector lens 42, and a light in the wavelength band transmitted through the first band-pass filter 43 is illuminated from below the sample 2. And a total-reflection mirror 44 for reflecting light to a mirror, and a condenser lens 45 for dark field suitable for fluorescence observation. The light transmitted through the condenser lens 45 illuminates the sample 2 from the back surface of the sample 2 arranged on the stage 3.

The first lamp 41 is a halogen lamp, and emits illumination light in a visible light region and a wavelength region including an excitation wavelength of the indocyanine green derivative-labeled antibody. The spectral transmission characteristics of the first band-pass filter 43 are as shown in FIG. 3. The first band-pass filter 43 transmits visible light and an excitation wavelength (770 to 790 nm) that excites an indocyanine green derivative-labeled antibody. Fluorescence wavelength of cyanine green derivative-labeled antibody (810-830n)
m).

The epi-illumination optical system 5 includes a second lamp 51 composed of a halogen lamp similar to the first lamp 41 that emits epi-illumination light, and collects illumination light from the second lamp 51. A second collector lens 52, a second band-pass filter 53 having the same characteristics as the first band-pass filter 43 transmitting a desired wavelength band of light transmitted through the second collector lens 52, The light in the wavelength band transmitted through the second band-pass filter 53 is reflected so as to be irradiated from the surface side of the sample 2, and
A first dichroic mirror 50 that transmits the fluorescence wavelength of the indocyanine green derivative-labeled antibody emitted from the sample 2.

The spectral transmission characteristics of the first dichroic mirror 50 are as shown in FIG. 4. The first dichroic mirror 50 transmits about 50% of visible light, reflects about 50%, and
It has the property of reflecting excitation light having a wavelength that excites the indocyanine green derivative-labeled antibody. In addition, the sample 2 has a property of almost transmitting a fluorescence wavelength emitted from the indocyanine green derivative-labeled antibody of the sample 2,
Either the first lamp 41 or the second lamp 51 is selected according to the sample 2 at the time of observation or the purpose of observation.

The visual observation system 6 is an objective lens 6 which is an observation optical system that captures the sample 2 placed on the stage 3.
1 and the optical path along which the optical image of the sample 2 travels,
Alternatively, an optical path conversion mirror 62, which is an optical path conversion means that can be inserted into and removed from the optical path for switching the direction of the imaging observation system 7, and an optical image reflected by the optical path conversion mirror 62 and displayed on a screen of a composite screen monitor described later. A prism 63 for reflecting the fluorescent image in a predetermined direction, and an eyepiece 64 to which an observer's eyes are put to observe the sample 2.

By inserting and removing the optical path conversion mirror 62, an optical image of the sample 2 transmitted through the first dichroic mirror 50 is transmitted in the direction of the second dichroic mirror. A half mirror that reflects a light image of the sample 2 transmitted through the mirror 50 toward the eyepiece lens 64 and transmits a fluorescent image displayed on the screen of the composite screen monitor, and a sample that transmits the first dichroic mirror 50 2 can be switched to a total reflection state in which the two light images are reflected toward the eyepiece lens 64 side.

The imaging observation system 7 converts an infrared light (fluorescence) component, which is light in a third wavelength band including at least a part of the fluorescence wavelength of the fluorescent substance, from the optical image of the sample 2 transmitted through the prism 63. The second dichroic mirror 7 is a light separating unit that reflects and transmits and spatially separates a visible light component which is light in a fourth wavelength band including at least a part of visible light.
0, a fluorescent image CCD 71 which is a first imaging means for imaging the fluorescent component reflected by the dichroic mirror 70 and converting the fluorescent component into an electric signal, and converting the visible light transmitted through the dichroic mirror 70 into an electric signal. A color CC in which a mosaic filter is arranged in front of the imaging surface as a second imaging unit to obtain a color image from a normal visible light component
D, a CCD 72 for visible light image, and each CCD 71, 72
A signal processing circuit 73 for generating an electric signal obtained by the photoelectric conversion in the above into a video signal, and an observation for receiving at least one of a visible light image and a fluorescent image on a monitor screen in response to the video signal generated by the signal processing circuit 73 And a composite screen monitor 75 that receives the video signal generated by the signal processing circuit 73, displays a fluorescent image, and composites and displays the fluorescent image during visual observation.

The composite screen monitor 75 is located near the prism 63 so that the fluorescent image displayed on the screen of the composite screen monitor 75 is reflected in the prism 63 and guided to the eyepiece lens 64. Are located in For this reason, the inspector operating the microscope needs the eyepiece 6.
The fluorescent image displayed on the screen of the composite screen monitor 75 can also be observed through 4.

The second dichroic mirror 7
The spectral transmission characteristic of No. 0 is as shown in FIG. 5, and has a characteristic of transmitting light in the visible light range and reflecting infrared light which is the fluorescence wavelength of the indocyanine green derivative-labeled antibody. That is, the second dichroic mirror 70 spatially separates the visible light component and the fluorescent light component.

The fluorescence component transmitted through the prism 63 and reflected by the second dichroic mirror 70 blocks the wavelength band of the excitation light included in the illumination light applied to the sample 2. As shown, the excitation light component of the indocyanine green derivative-labeled antibody is removed, and the excitation light cut filter 76 having a spectral transmission characteristic of transmitting a fluorescent component is provided.
, And is amplified by an image intensifier 77 having sensitivity to wavelengths around 350 nm to 910 nm, and forms an image on the imaging surface of the fluorescent image CCD 71.

Further, the CCDs 71 and 72 are synchronously driven by a CCD drive circuit (not shown) so that an image of 30 frames per second can be obtained from each of the CCDs 71 and 72.

The operation of the microscope device 1 configured as described above will be described. First, the second lamp 51 is turned on for observation with incident illumination light. Then, the lamp 51 emits illumination light in the visible light region and in the wavelength region including the excitation wavelength of the indocyanine green derivative-labeled antibody. This illumination light passes through the second collector lens 52 and the second bandpass filter 53, and then enters the first dichroic mirror 50. This first dichroic mirror 5
At 0, the excitation light at 770-790 nm and about 5
The sample 2 which reflects 0% of the light and irradiates the sample 2 previously stained with the indocyanine green derivative-labeled antibody through the objective lens 61 from the front side. When the first lamp 41 is selected as the lamp and observation is performed with transmitted illumination light, the illumination light emitted from the first lamp 41 is transmitted to the first collector lens 42 and the first bandpass filter. After passing through 43 and being reflected by a total reflection mirror 44, the sample 2 is irradiated from the back side through a condenser lens 45.

By irradiating the sample 2 with the illumination light, the visible light and the excitation light contained in the illumination light are absorbed and reflected by the sample 2 and are caused by the administered indocyanine green derivative-labeled antibody. Infrared light is emitted from the lesion tissue of the sample 2.

The light image including the reflected light and the infrared light emitted from the sample 2 is transmitted through the objective lens 61 and the first dichroic mirror 50, and the optical path conversion mirror 62 is pulled out to be in a completely transmitting state. And reaches the second dichroic mirror 70. Then, the optical image of the sample 2 is separated into a fluorescent (infrared light) component and a visible light (reflected light) component by the second dichroic mirror 70.

The fluorescence component reflected by the second dichroic mirror 70 passes through the excitation light cut filter 76, and the excitation light component of the indocyanine green derivative-labeled antibody is removed. The components are amplified by the image intensifier 77, imaged on the fluorescent image CCD 71, and photoelectrically converted. On the other hand, the second
The visible light component transmitted through the dichroic mirror 70 is
An image is formed on the visible light image CCD 72 and photoelectrically converted.

The electric signals photoelectrically converted by the CCDs 71 and 72 are input to a signal processing circuit 73 to generate image signals, and the image signals are output to a monitor 74 for observation and a monitor 75 for a composite screen, respectively. Display an optical image or fluorescent image.

On the screen of the observation monitor 74, one of a fluorescent image and a visible light image is displayed, or both images are simultaneously displayed in parallel or combined (superimposed) for diagnosis. Do.

Further, on the screen of the composite screen monitor 75, only the fluorescent image is displayed in response to the fluorescent image signal from the signal processing circuit 73. By inserting the optical path conversion mirror 62 to be in the half mirror state in the visual observation state, the optical image of the sample 2 is reflected by the optical path conversion mirror 62 and the prism 63 and guided to the eyepiece lens 64. The fluorescent image displayed on the composite screen monitor 75 is transmitted through the optical path conversion mirror 62 and is observed.
Observed at a position corresponding to the light image of FIG. That is, at the time of visual observation, it is possible to diagnose the presence or absence of cancer while associating the light image with the fluorescent image.

When only the naked eye observation is performed without observing the fluorescent image, the optical path conversion mirror 62 for total reflection is inserted so as to be located on the optical path, and the optical image of the sample 2 is observed with the naked eye. Do.

As described above, the illumination light of the lamp that emits the light of the first wavelength band including at least a part of the excitation wavelength of the fluorescent substance and the light of the second wavelength band including at least a part of the visible light is emitted from India. By irradiating a sample stained with a cyanine green derivative-labeled antibody, a light image containing reflected light and infrared light is emitted from this sample, and this light image is converted to a second image.
The dichroic mirror separates the light into a fluorescent (infrared light) component and a visible light (reflected light) component. By generating a signal, a fluorescence image and a visible light image can be obtained. This makes it easy to diagnose by observing the visible light image of the sample and the fluorescent image due to the infrared light caused by the administered indocyanine green derivative-labeled antibody.

Further, the light image from the sample is reflected toward the eyepiece lens in the optical path of the light image including the reflected light and the infrared light emitted from the sample, and the screen of the composite screen monitor arranged at a predetermined position is reflected. By arranging an optical path conversion mirror that makes a half mirror state that transmits the fluorescent image displayed above,
The diagnosis can be performed by overlapping the fluorescent image displayed on the composite screen monitor with the optical image of the sample reflected by the optical path conversion mirror. This allows the observer performing the microscopic inspection to simultaneously observe the fluorescence image and the light image of the sample to easily make a diagnosis.

In this embodiment, a mosaic filter is arranged in front of the CCD to detect normal light. However, a three-panel camera may be used to improve image quality.

Instead of using one kind of lamp as the lamp for the illumination light for observation, a combination of a plurality of light sources such as a halogen lamp for normal light observation and a semiconductor laser or a light emitting diode for exciting a fluorescent substance is used. It may be.

Further, an optical image of the fluorescent component is once imaged by a CCD, a video signal generated from the electric signal under photoelectric conversion is output to a monitor, and a displayed fluorescent image is observed through an eyepiece via a half mirror. Alternatively, the optical system may be configured so that the fluorescent screen of the image intensifier can be directly observed with the eyepiece.

7 to 9 relate to a second embodiment of the present invention, FIG. 7 is an explanatory view showing another schematic configuration of the entire microscope apparatus, FIG. 8 is an explanatory view showing the configuration of a rotary filter, and FIG. FIG. 4 is a characteristic diagram showing a spectral transmission characteristic of a rotation filter.

As shown in FIG. 7, in the microscope apparatus 10 of this embodiment, the illumination light of the visible light region and the wavelength region including the excitation wavelength of the indocyanine green derivative labeled antibody shown in the first embodiment are used as the epi-illumination system lamp. Instead of a halogen lamp that emits light, a laser light source 81 that emits near infrared light of 780 nm for exciting an indocyanine green derivative-labeled antibody is used. The sample 2
In order to observe the light image from the optical image, a rotating filter 83 provided with a plurality of filters that transmit each light component of the light image in an optical path in which the light image travels, and a motor 84 that drives the rotating filter 83 to rotate. C having a sensitivity from a visible light region to a near infrared light region for capturing a visible light image and an infrared light image transmitted through a filter provided in the rotary filter 83.
CD85.

As shown in FIG. 8, the rotary filter 83 is divided into four parts, and each part is provided with a filter corresponding to each light component. The four filters include a red transmission filter 83R transmitting red (R) light, a green transmission filter 83G transmitting green (G) light, a blue transmission filter 83B transmitting blue (B) light, and infrared light (IR). 9), and the spectral transmission characteristics of each of the filters 83R, 83G, 83B, 83F are as shown in FIG.

In the infrared transmission filter 83F, the wavelength for exciting the indocyanine green derivative-labeled antibody is cut off and the wavelength of fluorescence is transmitted. Reference numeral 82 denotes a diffusion lens 82 that diffuses laser light emitted from the laser light source 81. Other configurations are the same as those of the first embodiment, and the same members are denoted by the same reference numerals and description thereof will be omitted.

The operation of the microscope device 10 configured as described above will be described. In the present embodiment, when observing infrared light caused by the indocyanine green derivative-labeled antibody, both the first lamp 41 and the laser light source 81 are turned on, and when performing normal observation, the first lamp 41 is used. Just turn on and observe. Note that the rotary filter 83 is driven to rotate during both observations.

First, when fluorescence observation is required, the first
Lamp 41 and the laser light source 81 are turned on. Then, near-infrared light of 780 nm from the laser light source 81 passes through the diffusion lens 82 and is reflected by the first dichroic mirror 50 to irradiate the front side of the sample 2 through the objective lens 61 and to irradiate the sample 2 Illumination light from a lamp 41 composed of a halogen lamp is applied from the back side.
By using the laser light source 81, the sample 2 is irradiated with strong excitation light in addition to the excitation light from the first lamp 41.

When the illumination light is applied to the sample 2 from the two lamps, the reflected light and the fluorescent light, which are the optical images of the sample 2, pass through the objective lens 61 and the first dichroic mirror 50. , And enters the rotating filter 83 in a rotating state.

At this time, the rotation filter 83 is
4, the filters 83R, 83G, 83B, and 83F are sequentially disposed on the optical path, and are temporally separated into a visible light component and a fluorescent light component.

The filters 83R, 83R
The light transmitted through G, 83B, and 83F is imaged by the CCD 85. The CCD 85 is driven by a CCD driving circuit (not shown) in synchronization with the rotation of the rotary filter 83. Therefore, an electric signal of a red light image, an electric signal of a green light image, an electric signal of a blue light image, and an electric signal of an infrared light image are sequentially sent from the CCD 85 to the signal processing circuit 73. In this signal processing circuit 73, the CCD 85
After synchronizing the electric signals sent from the computer, a process for displaying an image corresponding to the observation monitor 74 and the composite screen monitor 75 is performed.

As described above, the rotary filter having the filters for transmitting the red light image, the green light image, the blue light image, and the infrared light image is disposed in the optical path of the light image of the sample. The light image transmitted through the camera is picked up by a CCD having sensitivity from the visible light region to the near-infrared light region and subjected to photoelectric conversion, whereby a visible light image (normal light image) is obtained through an observation monitor or an eyepiece. An infrared light image can be observed. Other functions and effects are the same as those of the first embodiment.

It should be noted that the present invention is not limited to only the above-described embodiments, and various modifications can be made without departing from the spirit of the invention.

[Appendix] According to the above-described embodiment of the present invention, the following configuration can be obtained.

(1) For a sample containing a fluorescent substance, light of a first wavelength band containing at least a part of the excitation wavelength of the fluorescent substance and light of a second wavelength band containing at least a part of visible light A light source means for irradiating light, a light emitted from the sample, a light in a third wavelength band including at least a part of a fluorescence wavelength of the fluorescent substance, and a fourth light including at least a part of visible light.
A light separating unit that spatially separates the light into light in a wavelength band;
A microscope apparatus comprising: an imaging unit; and a second imaging unit configured to image light including at least a part of the fourth wavelength band.

(2) The microscope apparatus according to Supplementary Note 1, wherein the light separating means is a dichroic mirror.

(3) The microscope apparatus according to Supplementary Note 1, further comprising an optical path conversion unit that can be inserted into and removed from the optical path of the light emitted from the sample and converts the optical path of the light.

(4) For a sample containing a fluorescent substance, light of a first wavelength band containing at least a part of the excitation wavelength of the fluorescent substance and light of a second wavelength band containing at least a part of visible light A light source means for irradiating light, a light emitted from the sample, a light in a third wavelength band including at least a part of a fluorescence wavelength of the fluorescent substance, and a fourth light including at least a part of visible light.
A light separating unit that temporally separates the light into a wavelength band light, and an imaging unit that captures light including at least a part of the third wavelength band and light including at least a part of the fourth wavelength band. Microscope equipment to be equipped.

(5) The microscope apparatus according to Supplementary Note 1 or 4, wherein the light source means is a transmission illumination optical system and an epi-illumination illumination optical system.

(6) The microscope apparatus according to Supplementary Note 5, wherein the light source means of the transmission illumination optical system and the light source means of the epi-illumination optical system are the same type of light source.

(7) The microscope device according to Supplementary Note 5, wherein the light source means of the transmission illumination optical system and the light source means of the epi-illumination optical system are composed of different types of light sources.

(8) The microscope apparatus according to Supplementary Note 4, wherein the light separating means is a rotary filter having a plurality of filters.

(9) The microscope apparatus according to Supplementary Note 1 or 4, further comprising: a visualization unit configured to make light including at least a part of the third wavelength band visible; and an image visualized by the visualization unit. Combining means for combining an image of light in the fourth wavelength band.

[0057]

As described above, according to the present invention, there is provided a microscope apparatus capable of simultaneously observing both a visible light image by ordinary observation light and a fluorescence image emitted from a body tissue by an indocyanine green derivative-labeled antibody. Can be.

[Brief description of the drawings]

FIGS. 1 to 6 relate to a first embodiment of the present invention, and FIG. 1 is an explanatory view showing a schematic configuration of an entire microscope apparatus.

FIG. 2 is a characteristic diagram for explaining excitation and fluorescence characteristics of an indocyanine green derivative-labeled antibody.

FIG. 3 is a characteristic diagram showing a spectral transmission characteristic of a bandpass filter.

FIG. 4 is a characteristic diagram showing a spectral transmission characteristic of a first dichroic mirror;

FIG. 5 is a characteristic diagram showing a spectral transmission characteristic of a second dichroic mirror;

FIG. 6 is a characteristic diagram showing a spectral transmission characteristic of an excitation light cut filter.

7 to 9 relate to a second embodiment of the present invention, and FIG. 7 is an explanatory view showing another schematic configuration of the entire microscope apparatus.

FIG. 8 is an explanatory diagram showing a configuration of a rotary filter.

FIG. 9 is a characteristic diagram showing a spectral transmission characteristic of a rotating filter.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... Microscope apparatus 2 ... Sample 41 ... First lamp 51 ... Second lamp 70 ... Second dichroic mirror 71 ... CCD for fluorescent image 72 ... CCD for visible light image 74 ... Monitor for observation 75 ... Monitor for synthetic screen

Claims (1)

[Claims] 1. A method according to claim 1, wherein a light in a first wavelength band including at least a part of an excitation wavelength of the fluorescent substance and a light in a second wavelength band including at least a part of visible light are applied to a sample including the fluorescent substance. Light source means for irradiating the light emitted from the sample into light in a third wavelength band including at least a part of the fluorescence wavelength of the fluorescent substance and light in a fourth wavelength band including at least a part of visible light; A light separating unit for separating light; a first imaging unit for capturing light including at least a part of the third wavelength band; and a second imaging unit for capturing light including at least a part of the fourth wavelength band. A microscope device, comprising: