JP2000321501A - Microscope using photoreflective optical material - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a microscope which eliminates the need for adjusting an aperture, lessens the effect of a culture vessel and allows the free change of a contrast and resolution by arranging the partial aperture near the pupil posi tion of an illumination optical system and arranging a modulation element having a photoreflective optical material near the pupil position of an image formation optical system conjugate with this pupil position. SOLUTION: The illumination optical system comprises a lens 12, a condenser lens 13 and a ring slit 14 near the pupil position and introduces the light of a light source 11 to a specimen 15. The image formation optical system comprises the objective 16 and the modulation element 17 having the photoreflective optical material (photochromic material) 17 arranged in the pupil position of the objective 16 conjugate with the pupil position of the illumination optical system. The image of the ring slit 14 by the zero order light past the aperture of the ring slit 14 is formed on a photoreflective optical element and the transmittance and refractive index of this portion alone change and can be observed as a phase difference microscope. Even more, the image formation position of the ring slit 14 acts as a phase film and, therefore, there is no need for adjusting the ring slit 14.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microscope using a photoreactive optical material.

[0002]

2. Description of the Related Art In a microscope, a colorless and transparent specimen such as cultured cells cannot be observed by ordinary bright-field observation because there is no difference in brightness. In order to observe such a colorless and transparent sample, an observation method such as a phase difference method and a modulation contrast method is used.

In a phase contrast microscope, a ring slit is arranged at a pupil position of an illumination system, and a phase plate having a phase film having a shape corresponding to the ring slit is arranged at a pupil position of an objective lens conjugate to the ring slit. The phase and intensity of the zero-order light of the light diffracted on the sample surface are changed, and the light is made to interfere with the diffracted light other than the zero-order light, so that the phase change of the sample is changed to a bright and dark contrast for observation.

Further, as disclosed in Japanese Patent Application Laid-Open No. 51-128548, a modulation contrast microscope has a rectangular aperture disposed at a pupil position of an illumination system, and furthermore, a pupil position of an objective lens conjugate to the rectangular aperture. A modulation plate having a modulation film in a region corresponding to a rectangular aperture and having a different transmittance is disposed, and the intensity of the diffracted light diffracted to one side with respect to the intensity of the zero-order light in the light diffracted on the sample surface is arranged The phase change of the sample is observed three-dimensionally (with a three-dimensional effect) by increasing the intensity and reducing the intensity of the diffracted light diffracted to the other.

In such a microscope, the illumination system between the pupil position of the illumination system in which the aperture is arranged and the pupil position of the objective lens in which the phase film and the modulation film are arranged, the lens in the objective lens, the aperture, Although there is little eccentricity in each part of the phase plate and the modulation plate, it is difficult to eliminate this eccentricity without adjustment. Depending on the amount of eccentricity, the zero-order light from the aperture may not enter the phase film and the modulation film. Therefore, conventional phase contrast microscopes and modulation contrast microscopes have a mechanism for decentering the aperture and adjusting the zero-order light from the aperture to enter the phase film and the modulation film.

In a phase contrast microscope, as described in Optics, Vol. 20, No. 9, pp. 590 to 594, the contrast and resolution are determined by the ratio between the phase film diameter and the pupil diameter. It is theoretically clear that the closer the ratio is to 1, the higher the resolution and the lower the contrast, and the closer the ratio between the phase film diameter and the pupil diameter to 0, the higher the contrast and the lower the resolution. Actually, since the optimum contrast and resolution differ depending on the specimen, the conventional phase contrast microscope is fixed to a phase film diameter that balances contrast and resolution.

Further, Japanese Patent Application Laid-Open No. 58-184115 and Japanese Patent Application Laid-Open No. 9-80313 disclose that the contrast which is most suitable for a specimen is changed by changing the transmittance of a phase film on a phase plate. Techniques to be obtained are described. That is, instead of the absorbing film on the phase plate, an electro-magnetic material whose transmittance changes by changing the applied voltage is used.

[0008] Further, there has been conventionally known a phase contrast microscope in which illumination is performed at different apertures without using a mechanical mechanism for changing the aperture. In this conventional example, as described in JP-A-5-333272 and JP-A-7-134250, a liquid crystal whose transmittance changes by changing an applied voltage instead of an aperture is used. is there.

[0009]

As described above, in the phase contrast microscope and the modulation contrast microscope, when the aperture adjustment is insufficient, all of the zero-order light does not enter the phase film or the modulation film. Therefore, sufficient contrast and three-dimensional effect cannot be obtained.

[0010] Further, the phase contrast microscope uses the NA of the objective lens.
Depending on the magnification and specifications of the objective lens, it is necessary to use different ring slits. In this case, if the lenses and the phase plates in the objective lenses are largely eccentric in different directions in the respective objective lenses, the aperture must be adjusted each time the objective lens to be used is switched.

Further, since it is difficult to change the diameter of the phase film after the phase film is once formed, a phase contrast microscope capable of freely changing the resolution and contrast has not been realized.

Further, when observing cells cultured in a culture vessel such as a well plate using a phase contrast microscope or a modulation contrast microscope, especially in the case of a culture vessel having a small diameter, the culture solution in the culture vessel is used. The surface is curved by the surface tension to form a lens, so that the projection position and the projection magnification between the opening and the phase plate or the modulation plate change, and the conjugate relationship between the opening and the phase plate or the modulation plate cannot be maintained. A similar situation occurs when observing cells located in the corners of a container even in a container having a small diameter. In such a state, all the zero-order light does not enter the phase film or the modulation film, so that the range in which light of the phase film or the modulation contrast can be obtained is narrowed, and the contrast is significantly reduced.

Among the above conventional examples, Japanese Patent Application Laid-Open No. 58-18411
Even if an electrochromic material is used as a modulating element as in a phase contrast microscope described in Japanese Patent Application Laid-Open No. H05-50013 and Japanese Patent Application Laid-Open No. 9-80313, the transmittance can be changed only in a region using the electrochromic material. Therefore, the adjustment of the opening is not required, and the influence of the curvature of the culture solution in the culture vessel cannot be reduced.

Further, in the phase contrast microscope described in JP-A-58-184115 and JP-A-9-80313, even if liquid crystal is used for the opening, the transmittance can be changed only in the divided area. In order to change the shape of the opening freely, the area must be divided innumerably,
It is virtually impossible. Also, it is not necessary to adjust the opening, and by changing the shape of the opening, it is possible to reduce the influence of the liquid level of the culture solution in the culture vessel, but the surface tension of the liquid level of the culture solution is reduced. In order to observe multiple containers because they differ depending on the shape of the culture container and the state of the culture solution,
The shape of the opening must be adjusted each time the container is changed. As described above, the conventional microscope cannot solve the above-mentioned disadvantage.

The present invention provides a microscope such as a phase-contrast microscope or a modulation contrast microscope that does not require adjustment of the aperture, reduces the influence of the culture vessel, and can freely change contrast and resolution.

[0016]

SUMMARY OF THE INVENTION A microscope according to the present invention has an illumination optical system for guiding light from a light source to a sample, and an imaging optical system, and has a partial aperture disposed near a pupil position of the illumination optical system. When,
A modulation element having a photoreactive optical material is arranged near a pupil position conjugate with a pupil position of the illumination optical system.

Further, the present invention relates to a microscope objective lens, and is characterized in that a modulation element having a photo-responsive optical material is arranged near a pupil position.

When light of a certain wavelength enters the photoresponsive optical element, the transmittance and the refractive index of the incident portion change reversibly. Here, the change in transmittance is a change in the direction in which the transmittance decreases when light is incident, compared to the transmittance when no light is incident. As this photoreactive optical element, "A glass handbook" published by Asakura Shoten Photochromic materials whose transmittance and refractive index change when light as described on pages 992 to 1000 are incident, and JP-A-7-316548.
There are photorefractive materials whose refractive index changes when light is incident thereon, as described in Japanese Patent No.

Since the microscope of the present invention has the above-described configuration, the portion where the zero-order light is incident from the aperture has a transmittance,
The refractive index changes. Also, other diffraction light diffracted by the sample is incident on a part of the modulation element other than the part where the zero-order light is incident. If the sample is sufficiently thin, the intensity is about 10% to 15% as compared with the zero-order light. Since it is small below, the transmittance and the refractive index of that portion do not change. In other words, the transmittance and the refractive index of the modulation element change only in the portion where the zero-order light is incident. Therefore, even if the opening is eccentric, a sufficient three-dimensional effect and contrast can be obtained, and there is no need to adjust the opening.

Further, due to the influence of the liquid level of the culture solution in the culture vessel, the projection position and the projection magnification between the opening and the phase plate are changed, and the position of the modulation element having the photoreactive optical material is changed to a different position.
Even if the next light passes, the transmittance and the refractive index of the passing portion change, so that the conventional phase contrast microscope, the aperture created by the modulation contrast microscope and the phase film on the modulation element,
It is possible to solve the disadvantage that the range in which the effect can be obtained is narrowed due to the mismatch of the modulation films, and the contrast is significantly reduced.

When the shape and position of the opening are changed, the transmittance and the refractive index of the portion corresponding to the opening in the modulation element are changed, so that the contrast, the resolution, and the appearance of the image can be freely changed.

It is difficult to adjust the components of the photoreactive optical material used in the microscope of the present invention so that changes in both the transmittance and the refractive index are optimal. It is difficult to obtain both the optimum change in transmittance and the optimum change in refractive index for a material.

Therefore, according to the present invention, the modulation element has a structure having a plurality of light-responsive optical materials, and the light-reactive optical material has a structure in which at least one of a change in transmittance and a change in refractive index is different. With such a configuration of the modulation element, the optimum transmittance and refractive index can be easily obtained. In addition, if the modulation element is formed of a plurality of different regions of at least one of the transmittance change amount and the refractive index change amount different photoreactive material, different contrast and appearance can be obtained by switching the aperture. .

Also, the phase contrast microscope sets the phase of the zero-order light to λ
The phase difference between the 0th-order light and the 1st-order light is set to λ / 2 by advancing / 4, and the dark contrast that makes the phase object look dark, and the 0th-order light and the 1st-order light by delaying the position of the 0th-order light by λ / 4. There is a bright contrast that makes the phase object look brighter in phase. Here, in the case of dark contrast, the phase of the zero-order light is λ / 4
In order to proceed, the refractive index of the portion where the zero-order light enters must be lower than the refractive index of the other portions where the diffracted light enters.

However, in the photoreactive optical material, it is difficult to lower the refractive index of the portion to which the zero-order light is applied to such an extent that the phase of the zero-order light is advanced by λ / 4 compared with the other portions.

The microscope of the present invention has at least one photo-responsive optical material whose transmittance changes, and is arranged in an illumination optical system and has a shape corresponding to a partial aperture. The photoreactive optical material is formed of a low refractive index material having a refractive index of 1.5 or less, and is bonded to another glass substrate. As a result, the portion other than the low-refractive-index material is filled with the adhesive, and the low-refractive-index material (refractive index 1.5 or less) and the filled bonding agent (refractive index 1.
The phase difference of the zero-order light can be easily advanced by λ / 4 due to the difference in the refractive index from that of (5 to 1.6).

Further, as described in the above-mentioned JP-A-51-128548, the modulation contrast microscope uses the intensity with respect to the intensity of the zero-order light among the light diffracted on the sample surface, while the intensity of the diffracted light as one. It is necessary to increase the intensity of the diffracted light on the other hand. Therefore, a region having a transmittance lower than that of the portion where the zero-order light is applied is required. However, since the transmittance of only the portion where light is incident on the photoreactive material changes, it is not possible to obtain a region having a low transmittance other than the portion on which the light is incident only with the photoreactive optical material.

In the microscope according to the present invention, the modulation element has a photoreactive optical material in which at least one of the transmittance and the refractive index changes, and is arranged in a partial region of the modulation element. It consisted of a light absorbing material having a lower transmittance than the photoreactive optical material after the change. With such a configuration, it is possible to obtain a portion having a lower transmittance than a portion where the zero-order light is incident.

The photochromic material, which is one of the photoreactive optical materials, can change the wavelength at which the transmittance changes depending on the substance contained therein, as described on page 996 of the above-mentioned document.

The microscope of the present invention has a light source, an illumination optical system for guiding the light from the light source to the sample, and an imaging optical system, and a part of the wavelength within the illumination optical system or the imaging optical system. A filter that cuts light is removably arranged, or an interference filter formed by a dielectric multilayer film is inclined with respect to the optical axis and arranged so that the inclination angle can be changed. It is configured to have a plurality of different photoreactive optical materials. With such a configuration, the amount of illumination light can be adjusted.

The microscope has a light source, an illumination optical system for guiding the light from the light source to the sample, and an imaging optical system, and a filter for cutting a part of the wavelength can be attached to and detached from the illumination optical system. Or an interference filter composed of a dielectric multilayer film is arranged so that the inclination angle with respect to the optical axis can be changed, a partial aperture is arranged near the pupil position of the illumination optical system, and the pupil of the illumination optical system is arranged. In the vicinity of the pupil position of the imaging optical system conjugate with the position, a modulation element having a plurality of photoresponsive optical materials having different wavelengths at which the transmittance changes is arranged.
With such a configuration, it is possible to change the wavelength to be cut, and a plurality of photoreactive optical materials having different wavelengths at which the transmittance changes can change the transmittance only at a certain wavelength. Or the type of material can be changed so that the transmittance of all wavelengths changes. As a result, the intensity of the zero-order light can be freely changed, and observation with different contrast suitable for various different specimens becomes possible.

[0032]

Next, an embodiment of the microscope according to the present invention will be described.

FIG. 1 shows a microscope according to a first embodiment of the present invention. A light source 11 and a lens 12 for introducing light from the light source 11 are shown.
, A condenser lens 13, and a ring slit 14 disposed near the pupil position of the illumination optical system. The illumination optical system guides the light of the light source to the sample 15, and is conjugate with the pupil position of the objective lens 16 and the illumination optical system. And a modulating element 17 having a photochromic material 17a, which is one of the photoreactive optical materials, disposed at the pupil position of a simple objective lens.

In the microscope shown in FIG. 1, the zero-order light passing through the ring slit 14 is a photochromic material which is one of the photoreactive optical materials placed at the pupil position of the objective lens 16 by the condenser lens 13 and the objective lens 16. Material 17a
Is formed on the modulation element 17 having

According to the microscope of this embodiment, since the image of the ring slit by the 0th-order light passing through the opening of the ring slit 14 is formed on the photoreactive optical element, the transmittance and the refractive index of only that portion are formed. Changes and can be observed as a phase contrast microscope. In addition, since the imaging position of the ring slit by the zero-order light acts as a phase film, there is no need to adjust the ring slit. In addition, phase difference observation can be performed without being affected by a culture vessel or the like.

FIG. 2 shows a second embodiment of the present invention.
The microscope according to this embodiment includes a light source 21, a lens 22 for introducing light from the light source 21, a condenser lens 23, and a partial aperture 24 arranged near a pupil position of the illumination optical system. An illumination optical system for guiding the specimen, an objective lens 26, and an imaging optical system including a modulation element 27 having a photochromic material 27a at a pupil position of the objective lens which is a position conjugate with a pupil position of the illumination optical system. ing.

In this microscope, the zero-order light passing through the partial aperture 24 is imaged by the condenser 23 and the objective lens 26 on a modulation element having a photo-responsive optical material at the pupil position of the objective lens.

In the microscope according to this embodiment, the partial aperture 24 is formed eccentrically as shown in the figure, whereby the zero-order light passing through the partial aperture is obliquely incident on the sample, whereby the sample The phase change can be observed three-dimensionally. Further, if a partial aperture is formed so as to be close to the outer periphery of the pupil of the objective lens at an angle close to the NA of the objective lens, one of the diffracted lights enters the outer periphery of the pupil, so that the same configuration as the modulation contrast is obtained. A three-dimensional effect can be obtained.

The microscope according to this embodiment also has the modulation optical element 2
An image of the partial aperture 24 is formed on the reference numeral 7, and the transmittance or the refractive index of the image of the partial aperture irradiated with the zero-order light changes. Therefore, the adjustment of the partial opening is unnecessary.

FIG. 3 is a diagram showing a configuration of a microscope according to the third embodiment of the present invention. This microscope comprises a light source 31, a lens 32 for introducing light from the light source, a condenser lens 33, and a ring slit 34 arranged near the pupil position of the illumination optical system. An illumination optical system leading to an objective lens 35; an objective lens 36; and a relay lens 37 for relaying an image formed by the objective lens.
And an imaging optical system including a modulation element 38 having a photochromic material 38a disposed at a pupil position of the relay lens conjugate with the pupil position of the illumination optical system and the pupil position of the objective lens.

In the microscope of this embodiment, the modulation element is arranged at the pupil position of the relay lens conjugate to the pupil position of the illumination optical system and the pupil position of the objective lens.

When the modulation element is arranged in the objective lens, the objective lens must be disassembled in order to replace the modulation element having different characteristics, and the replacement is troublesome. If the modulation element is arranged at the pupil position of the lens, the replacement can be facilitated.

If a partial aperture as in the second embodiment shown in FIG. 2 is used instead of the ring slit, stereoscopic observation is possible.

FIG. 4 shows a first example of a modulation element used in the microscope of the present invention. This modulation element is a glass substrate 41
A photochromic material 42 having a small phase change with respect to a change in density and a photochromic material 43 having a large change in refractive index with respect to a change in transmittance are arranged above.

In this modulator, the thickness of the photochromic material 42 having a small change in refractive index with respect to the change in transmittance and the thickness of the photochromic material 43 having a large change in refractive index with respect to the change in transmittance are changed. Thereby, the optimum transmittance and refractive index can be easily obtained.

FIG. 5 shows a second example of the modulation element used in the microscope of the present invention. The modulation element shown in FIG. 5 includes a photochromic material 52 having a large change in transmittance disposed in a partial region on a glass substrate 51 and a photochromic material having a small change in transmittance disposed in another partial region. Material 53
It is composed of

In the modulation device shown in FIG. 5, the contrast can be easily changed by changing the aperture to change the region where the zero-order light is incident.

FIG. 6 shows a third example of the modulation element used in the microscope of the present invention.

The modulation element shown in FIG.
1 and a photorefractive material 63.

In the element shown in FIG. 6, the transmittance is changed by the photochromic material 62, and the refractive index is changed by the photorefractive material 63, so that the optimum transmittance is changed. Can be easily obtained.

FIG. 7 shows a fourth example of a modulation element used in the microscope of the present invention, which is a phase difference modulation element. In this modulation element, a photochromic material 72 is disposed on a glass substrate 71, and a ring-shaped magnesium fluoride (MgF ₂ ) 73 corresponding to a ring slit disposed in an illumination optical system of a phase microscope is disposed. The glass substrate 74 is bonded by an adhesive 75. The refractive index of magnesium fluoride is 1.37, and the refractive index of the adhesive is 1.5 to
1.6.

By using this modulation element, 0
The phase of the next light can be advanced by λ / 4 from the other diffracted lights, and the phase difference can be observed with dark contrast.

FIG. 8 shows a fifth example of a modulation element used in the microscope of the present invention, which is a modulation contrast modulation element.

This modulating element is composed of a photochromic material 82 disposed on a glass substrate 81 and a light absorbing material 83 on another glass substrate 84 disposed in a region close to the photochromic material 82. I have. Reference numeral 85 denotes an adhesive.

By using this modulation element, a region having a lower transmittance than the portion where the zero-order light is incident can be obtained, and it is possible to easily observe the modulation contrast. Here, the transmittance of the portion where the zero-order light is incident is preferably 10% to 15%. The transmittance of the light absorbing material is best at 0%, but a sufficient effect can be obtained if the transmittance of the light absorbing material is smaller than the transmittance of the portion where the zero-order light is incident.

FIG. 9 is a diagram showing the configuration of the illumination system of the microscope according to the present invention. A light source 91, a lens 92 for introducing light from the light source, a condenser lens 93, and a variable inclination angle with respect to the optical axis are arranged. A short-wavelength cut filter 94 made of a dielectric multilayer film, a photochromic material 95a having a spectral transmittance characteristic whose transmittance changes at 450 nm or less as shown by a curve a in FIG. 10, and a curve b in FIG. The modulation device 95 includes the photochromic material 95b whose transmittance changes at 500 nm or less. Reference numeral 96 denotes, for example, a specimen.

According to this illumination system, the wavelength to be cut is shifted to the longer wavelength side by tilting the short-wavelength cut filter made of the dielectric multilayer film to change the transmittance of the photochromic material, thereby changing the amount of illumination light. It is like that.

FIG. 11 shows a microscope according to a fourth embodiment of the present invention. This microscope includes a light source 111, a lens 112 for introducing light from the light source 111, a condenser lens 113, a ring slit 114 disposed near a pupil position of the illumination optical system, and a tilt angle with respect to the optical axis. And a short wavelength cut filter 115 having a dielectric multilayer film arranged so as to obtain light from a light source.
Optical system, an objective lens 117, and photochromic materials 118a and 5a whose transmittance changes at 450 nm or less at the pupil position of the objective lens conjugate with the pupil position of the illumination optical system.
The imaging optical system is composed of a modulation element 118 having a photochromic material 118b whose transmittance changes below 00 nm.

In the microscope according to this embodiment, the wavelength to be cut is shifted to the longer wavelength side by tilting a short wavelength cut filter made of a dielectric multilayer film to change the transmittance of the photochromic material, thereby changing two different types of contrast. Can be obtained. Further, more kinds of contrast can be obtained by increasing the number of photochromic materials whose transmittance changes at different wavelengths.

The microscope or microscope objective of the present invention comprises:
It has the features as described above, and the following configurations can also achieve the object in addition to the configurations described in the claims. (1) The microscope according to claim 1 or the objective lens according to claim 2, wherein the modulation element is disposed in the illumination optical system, and the modulation is performed. The element has at least one photoreactive optical material having a shape corresponding to the shape of the partial opening and having a variable transmittance, and the photoreactive optical material is a low refractive index material having a refractive index of 1.5 or less. A microscope or a microscope objective lens, comprising:

(2) A photo-responsive optical material having at least one of a transmittance and a refractive index that changes with a modulation element provided in claim 1 or claim 2. A modulation element comprising a light-absorbing material having a transmittance lower than the transmittance of the photoreactive optical material after the change.

(3) A light source, a microscope optical system having an illumination optical system for guiding the light from the light source to the sample, and an imaging optical system, are removably disposed in the microscope optical system. Having a filter that cuts some wavelengths or an interference filter formed by a dielectric multilayer film that can change the tilt angle with respect to the optical axis, and a plurality of photoreactive optical materials with different wavelengths at which the transmittance changes Microscope.

(4) In the microscope according to the first aspect of the present invention, the inclination angle with respect to the optical axis is changed with respect to a filter or a filter that cuts a part of the wavelength and that is detachably mounted on the illumination optical system. A microscope having an interference filter formed by a dielectric multilayer film arranged so as to obtain a light-sensitive optical material having different wavelengths at which transmittance changes. .

(5) The objective lens according to claim 2, wherein the modulation element has a plurality of photo-reactive optical materials, and the photo-reactive optical materials have a change in transmittance or a refraction. An objective lens characterized in that at least one of the amounts of change in the rate is different.

[0065]

According to the present invention, there is no need to adjust the aperture, the influence of the culture vessel is reduced, and the contrast and resolution can be freely changed.

[Brief description of the drawings]

FIG. 1 is a diagram showing a first embodiment of a microscope of the present invention.

FIG. 2 is a diagram showing a second embodiment of the microscope of the present invention.

FIG. 3 is a diagram showing a third embodiment of the microscope of the present invention.

FIG. 4 is a diagram showing a first example of a modulation element used in the microscope of the present invention.

FIG. 5 is a diagram showing a second example of a modulation element used in the microscope of the present invention.

FIG. 6 is a diagram showing a third example of a modulation element used in the microscope of the present invention.

FIG. 7 is a diagram showing a fourth example of a modulation element used in the microscope of the present invention.

FIG. 8 is a diagram showing a fifth example of a modulation element used in the microscope of the present invention.

FIG. 9 is a diagram showing a configuration of an illumination system used in the microscope according to the present invention, in which the amount of illumination can be changed.

10 is a diagram showing an outline of spectral transmittance characteristics of a photochromic material used in the illumination system shown in FIG. 9;

FIG. 11 is a diagram showing a fourth embodiment of the microscope of the present invention.

[Explanation of symbols]

11, 21, 31, 91, 111 ... light source 12, 22, 32, 92, 112 ... lens 13, 23, 33, 93, 113 ... condenser lens 14, 34, 114 ... ring slit 24: Partial aperture 16, 26, 36, 117 ... Objective lens 17, 27, 38, 118 ... Modulating element

Claims (3)

[Claims] 1. An illumination optical system comprising: a light source; an illumination optical system for guiding light from the light source to a sample; and an imaging optical system; a partial aperture disposed near a pupil position of the illumination optical system; A microscope having a modulation element disposed in the vicinity of a pupil position of the imaging optical system conjugate with the pupil position of the imaging element, wherein the modulation element has a photoreactive optical material. 2. A microscope objective lens in which a modulation element having a photoreactive optical material is arranged near a pupil position. 3. The optical modulator according to claim 2, wherein the modulating element has a plurality of photo-responsive optical materials, and the photo-responsive optical materials differ from each other in at least one of a change in transmittance and a change in refractive index. 1 microscope.