JP5829031B2 - Faint light observation microscope and faint light acquisition method - Google Patents

Description

  The present invention relates to a weak light observation microscope and a weak light acquisition method for acquiring and observing weak light emitted from an observation target.

Conventionally, as observation of weak light, for example, observation of a change in the fine structure level of a cell or tissue for a long time using a light emission microscope is performed.
For example, as shown in FIG. 5, the light emission microscope includes an imaging optical system 51 including an objective lens 51a, an imaging lens 51b, and an imaging element 51c. In FIG. 5, reference numeral 60 denotes a sample such as a cell / tissue as an observation target, 60 a denotes a light emitting portion of the observation target 60, 61 denotes a petri dish that houses the observation target 60, a slide glass, and a micro A container 60 a ′ such as a plate is an image of the light emitting unit 60 a ′ picked up by the image pickup device 51. Then, in the observation using the light emission microscope, the weak light of a predetermined wavelength emitted from the observation target by chemical reaction with light emission reagent such as a predetermined time (e.g., 15 minutes, etc.) it is exposed, by repeating this dozens of times Capture changes in tissues and cells.
Conventionally, as this type of light emission microscope, for example, those described in the following Patent Documents 1 and 2 have been proposed.

JP 2007-108154 A WO07 / 074923 Publication

  In general, when a light emission image from an observation target is imaged using a light emission microscope, there is a method of increasing the amount of light received by increasing the exposure time of the image sensor in order to compensate for weak light emission intensity. Used. Further, in order to obtain a weak light emission image as brightly as possible, a high NA lens is used for the objective lens and the imaging lens.

Here, the larger the NA of the objective lens, the brighter the emission image.
However, when observing a wide range with a low-magnification objective lens, it is difficult to increase the NA. Further, there is a limit to the NA even when a high-magnification objective lens is used.
For this reason, in the conventional light-emitting microscope, it is necessary to compensate for the lack of brightness by increasing the exposure time of the image sensor as described above. However, when the exposure time of the image sensor is long, it is difficult to capture a fast change of the observation target.

By the way, Patent Document 2 discloses a configuration in which a unit that captures a light emission image and a unit that captures a fluorescence image are arranged to face each other with respect to an observation target.
Here, the present applicant applies the above-described configuration disclosed in Patent Document 2, and arranges a unit that captures two luminescent images to face an observation target, and synthesizes the luminescent images acquired by the two units. The idea was to compensate for the emission intensity.

  However, even if the units for capturing two luminescent images are arranged opposite to each other, the luminescence intensity captured by each unit is weak, and the exposure time of the image sensor in each unit cannot be shortened. The problem that it is difficult to photograph changes cannot be solved.

  The present invention has been made in view of such conventional problems, and is capable of shortening the exposure time of the imaging device, using a low-magnification lens, and capable of photographing a fast change of an observation object. It aims at providing the microscope and the weak light acquisition method.

In order to achieve the above object, a weak light observation microscope and a weak light acquisition method according to the present invention have an imaging optical system including an objective lens, an imaging lens, and an imaging device, and are observed through the imaging optical system. A weak light observation microscope that acquires predetermined weak light emitted from an object, the second objective lens, a second imaging lens, and the predetermined weak light disposed at a position conjugate with the observation object A light return optical system having a mirror that reflects the wavelength of light and transmits a wavelength other than the wavelength of the predetermined weak light, on the opposite side of the imaging optical system with respect to the observation target, and An illumination optical system that is provided on the opposite side of the mirror from the second imaging lens and emits illumination light having a wavelength different from the wavelength of the predetermined weak light toward the mirror. It is characterized by.

  Further, in the feeble light observation microscope of the present invention, the imaging optical system further includes a wavelength selection element that blocks the wavelength of the illumination light and transmits the wavelength of the predetermined feeble light. Is preferred.

  In the weak light observation microscope of the present invention, it is preferable that an aberration correction unit is provided at the pupil position of the second objective lens.

  In the feeble light observation microscope of the present invention, it is preferable that the aberration correction unit is a correction ring.

  In the feeble light observation microscope of the present invention, it is preferable that the aberration correction unit is a phase modulator.

  In the weak light observation microscope of the present invention, it is preferable that the predetermined weak light is light emitted from the observation object by a chemical reaction.

  In the feeble light observation microscope of the present invention, the predetermined feeble light is fluorescence emitted from the observation object by irradiation with an excitation wavelength, and the illumination light has an excitation wavelength that excites the observation object. Is preferred.

  ADVANTAGE OF THE INVENTION According to this invention, the exposure time of an image pick-up element can be shortened, the use of a low magnification lens, and the weak light observation microscope and weak light acquisition method which can image | photograph the rapid change of an observation object are obtained.

It is explanatory drawing which shows the principal part structure of the weak light observation microscope concerning 1st Embodiment of this invention. It is explanatory drawing which shows the principal part structure of the weak light observation microscope concerning 2nd Embodiment of this invention. It is explanatory drawing which shows the principal part structure of the weak light observation microscope concerning 3rd Embodiment of this invention. It is explanatory drawing which shows the principal part structure of the weak light observation microscope concerning 4th Embodiment of this invention. It is explanatory drawing which shows the example of 1 structure of the conventional light emission microscope.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. Of the embodiments of the present invention, the first to third embodiments are reference examples of the present invention.
First Embodiment FIG. 1 is an explanatory view showing the main configuration of a weak light observation microscope according to a first embodiment of the present invention.
The weak light observation microscope of the present embodiment includes an imaging optical system 1 and a light returning optical system 2.
The imaging optical system 1 includes an objective lens 1a, an imaging lens 1b, and an image sensor 1c.
The optical returning optical system 2 includes a second objective lens 2a, a second imaging lens 2b, and a plane mirror 2c, and is provided on the opposite side of the imaging optical system 1 with respect to the observation target 10. ing.
The mirror 2c is disposed at a position conjugate with the observation object 10.
The observation object 10 is a cell / tissue that includes a light emitting portion 10a that emits predetermined weak light by a chemical reaction with a luminescent reagent or the like, and uses a sample that transmits light with respect to the wavelength of the predetermined weak light. ing. The observation object 10 is accommodated in a container 11 such as a petri dish, a slide glass, or a microplate. The container 11 is placed on a stage (not shown).
The objective lens 1a and the second objective lens 2a can adjust the focus with respect to the observation object 10 by changing the relative distance between the objective lens 1a and the second objective lens 2a.

In the weak light observation microscope of the first embodiment configured as described above, the predetermined weak light emitted from the light emitting unit 10a of the observation target 10 toward the imaging optical system 1 side is the objective lens 1a and the imaging lens. Through 1b, an image is formed on the imaging surface of the imaging device 1c. In FIG. 1, 10a ′ is an image of the light emitting unit 10 formed on the imaging surface of the imaging device 1c.
On the other hand, predetermined weak light emitted from the light emitting unit 10a of the observation target 10 toward the light return optical system 2 side passes through the second objective lens 2a and the second imaging lens 2b, and then the surface of the mirror 2c. , Reflected by the mirror 2c, and returned to the observation object 10 through the second imaging lens 2b and the second objective lens 2a. In FIG. 1, 10a ″ is an image of the light emitting portion 10a formed on the surface of the mirror 2c. The predetermined weak light returned to the observation target 10 further directs the observation target 10 toward the imaging optical system 1 side. And passes through the objective lens 1a and the imaging lens 1b to form an image on the imaging surface of the imaging device 1c.
As a result, predetermined weak light having an amount of light twice as large as that of the conventional weak light observation microscope is incident on the image pickup device 1c, and the exposure time of the image pickup device 1c can be reduced to about half.
As a result, it is possible to capture a light emission image even using a low-magnification objective lens that cannot increase the NA, and it is possible to capture a fast change in the observation target (light emitting portion).

Second Embodiment FIG. 2 is an explanatory view showing the main configuration of a weak light observation microscope according to a second embodiment of the present invention. In addition, about the same structural member as the weak light observation microscope of 1st Embodiment, the same code | symbol is attached | subjected and detailed description is abbreviate | omitted.
In the weak light observation microscope of the present embodiment, the mirror disposed at a position conjugate with the observation object 10 in the light return optical system 2 is constituted by a deformable mirror 2c ′. Other configurations are substantially the same as the weak light observation microscope of the first embodiment.

In the configuration in which the planar mirror 2c is used as a mirror disposed at a position conjugate with the observation target 10 in the optical return optical system 2, when the thickness of the observation target 10 is not constant, a part of the light emitting units 10a of the observation target 10 is used. The predetermined weak light emitted toward the imaging optical system 1 side forms an image at a position before and after the plane mirror 2c. Then, the light reflected by the flat mirror 2c does not return to the light emitting point of the light emitting unit 10a in the observation target 10, but forms an image at a position before and after the light emitting point. The light is transmitted toward the imaging optical system side and enters the imaging surface of the imaging element 1c without passing through the objective lens 1a and the imaging lens 1b. As a result, an image captured by the image sensor 1 is blurred.
However, according to the feeble light observation microscope of the second embodiment, the reflecting surface of the deformable mirror 2c ′ is deformed in accordance with the thickness or the optical path length of the observation object 10, so that the predetermined light reflected by the deformable mirror 2c ′ is obtained. Can be accurately returned to the position of the light emitting point of the observation object 10. For this reason, even when the thickness of the observation object 10 is not constant, it is possible to prevent the image from being blurred.
Other functions and effects are substantially the same as those of the weak light observation microscope of the first embodiment.

Third Embodiment FIG. 3 is an explanatory view showing the main configuration of a weak light observation microscope according to a third embodiment of the present invention. In addition, about the same structural member as the weak light observation microscope of 2nd Embodiment, the same code | symbol is attached | subjected and detailed description is abbreviate | omitted.
The weak light observation microscope of the present embodiment further includes an optical path dividing member 3 and an illumination optical system 4 in addition to the configuration of the second embodiment.
The optical path dividing member 3 is composed of a dichroic mirror having optical characteristics that transmits a predetermined weak light wavelength and reflects a wavelength other than the predetermined weak light wavelength, and the second objective lens 2a and the second imaging lens. It arrange | positions between the lenses 2b.
The illumination optical system 4 is configured to emit illumination light having a wavelength different from a predetermined weak light wavelength toward the optical path dividing member 3, and is provided on the side of the light return optical system 2.
In the example of FIG. 3, the illumination optical system 4 includes a light source 4a, a collimator lens 4b that converts light from the light source into a parallel light beam, and a bandpass filter 4c. The light source 4a emits normal white light. The bandpass filter 4c has an optical characteristic of shielding a predetermined weak light wavelength and transmitting a wavelength other than the predetermined weak light wavelength. The band pass filter 4c is provided in the optical path of the illumination optical system 4 so that it can be inserted and removed.

The imaging optical system 1 further includes an illumination light cut filter 1d that can be inserted and removed between the objective lens 1a and the imaging lens 1b. The illumination light cut filter 1d has a characteristic of blocking the wavelength of illumination light and transmitting the wavelength of predetermined weak light.
Other configurations are substantially the same as the weak light observation microscope of the second embodiment.

In the feeble light observation microscope of the third embodiment configured as described above, the feeble light observation and the transmitted illumination observation with normal light can be switched by the following two methods.
In the first method, the illumination light cut filter 1d is inserted between the objective lens 1a and the imaging lens 1b and the light source 4a of the illumination optical system 4 is turned on to form an image with the objective lens 1a. In this method, the illumination light cut filter 1d is inserted into and removed from the lens 1b.
When faint light observation is performed, an illumination light cut filter 1d is inserted. Predetermined weak light emitted from the observation target 10 toward the light return optical system 2 side passes through the second objective lens 2a, passes through the optical path dividing member 3, passes through the second imaging lens 2b, and The image is formed on the surface of the mirror 2c, reflected by the mirror 2c, transmitted through the optical path dividing member 3 through the second imaging lens 2b, and returned to the observation object 10 through the second objective lens 2a. Further, the predetermined weak light transmitted through the observation target 10 toward the imaging optical system 1 side and the predetermined weak light emitted from the observation target 10 toward the imaging optical system 1 side pass through the objective lens 1a. Then, the light passes through the illumination light cut filter 1d, passes through the imaging lens 1b, and forms an image on the imaging surface of the imaging device 1c. The illumination light from the illumination optical system 4 is reflected by the optical path dividing member 3 and enters the second objective lens 2a and the observation object 10. The light transmitted through the observation object 10 enters the illumination light cut filter 1d through the objective lens 1a, but is blocked by the illumination light cut filter 1d.
On the other hand, when performing transmitted illumination observation with normal light, the illumination light cut filter 1d is extracted. The illumination light from the illumination optical system 4 is reflected by the optical path dividing member 3 and enters the observation object 10 through the objective lens 2a. The light transmitted through the observation object 10 forms an image on the imaging surface of the imaging element 1c through the objective lens 1a and the imaging lens 1b.

The second method is a method of switching ON / OFF of the light source 4a of the illumination optical system 4 in a state where the illumination light cut filter 1d is extracted from between the objective lens 1a and the imaging lens 1b.
When performing faint light observation, the light source 4a of the illumination optical system 4 is turned off. Predetermined weak light emitted from the observation target 10 toward the light return optical system 2 side passes through the second objective lens 2a, passes through the optical path dividing member 3, passes through the second imaging lens 2b, and The image is formed on the surface of the mirror 2c, reflected by the mirror 2c, transmitted through the optical path dividing member 3 through the second imaging lens 2b, and returned to the observation object 10 through the second objective lens 2a. The predetermined weak light transmitted through the observation target 10 toward the imaging optical system 1 side and the predetermined weak light emitted from the observation target 10 toward the imaging optical system 1 side are connected to the objective lens 1a. An image is formed on the imaging surface of the imaging element 1c through the image lens 1b.
On the other hand, when performing transmission illumination observation with normal light, the light source 4a of the illumination optical system 4 is turned on. The illumination light from the illumination optical system 4 follows the same path as in the first method, and forms an image on the imaging surface of the imaging element 1c.

  According to the weak light observation microscope of the third embodiment, the weak light observation and the transmitted illumination observation can be switched, and the same effect as the weak light observation microscope of the first and second embodiments can be obtained. That is, a predetermined weak light having a light quantity about twice that of a conventional weak light observation microscope is incident, and the exposure time of the image sensor 1c can be shortened to about half, and the low magnification objective lens that cannot increase the NA. It is possible to pick up a luminescent image even when using, and it is possible to take a quick change of the observation target (light emitting portion), and the reflection surface of the deformable mirror 2c ′ is to be observed. The predetermined weak light reflected by the deformable mirror 2c ′ can be accurately returned to the position of the light emitting point of the observation object 10 by being deformed according to the thickness of 10 or the optical path length. Even if is not constant, it is possible to prevent image blurring.

In the example of FIG. 3, the predetermined weak wavelength is described as the emission wavelength emitted from the observation target by a chemical reaction with the luminescent reagent or the like, but the predetermined weak wavelength is emitted from the observation target by irradiation with the excitation wavelength. It may be a fluorescence wavelength.
In that case, a band pass filter 4c may be used that has an optical characteristic of shielding a predetermined fluorescence wavelength and transmitting a predetermined excitation light wavelength.
In the third embodiment, the mirror disposed at a position conjugate with the observation target 10 in the light return optical system 2 is configured by the deformable mirror 2c ′, but may be configured by the plane mirror 2c.

Fourth Embodiment FIG. 4 is an explanatory view showing the main configuration of a weak light observation microscope according to a fourth embodiment of the present invention. In addition, about the same structural member as the weak light observation microscope of 1st Embodiment, the same code | symbol is attached | subjected and detailed description is abbreviate | omitted.
In the feeble light observation microscope according to the present embodiment, a mirror disposed at a position conjugate with the observation target 10 in the light returning optical system 2 reflects a predetermined weak light wavelength and transmits a wavelength other than the predetermined weak light wavelength. It is composed of a dichroic mirror 2c ″ that also has a function as an optical path dividing member having the above characteristics.
In addition to the configuration of the first embodiment, the weak light observation microscope of the present embodiment further includes an illumination optical system 4 ′ on the side opposite to the second imaging lens 2b with respect to the dichroic mirror 2c ″. Have.
In the example of FIG. 4, the illumination optical system 4 ′ includes a light source 4a ′ and a condensing lens 4b ′ for condensing the light from the light source 4a ′ on the dichroic mirror 2c ″. The light source 4a ′ is a predetermined light source 4a ′. Illumination light having a wavelength different from the weak light wavelength is emitted, instead of the light source 4a ′ in the illumination optical system 4 ′, a light source 4a that emits normal white light similar to the example of FIG. A band pass filter 4c may be provided.

The imaging optical system 1 further includes an illumination light cut filter 1d that can be inserted and removed between the objective lens 1a and the imaging lens 1b. The illumination light cut filter 1d has a characteristic of blocking the wavelength of illumination light and transmitting the wavelength of predetermined weak light.
Other configurations are substantially the same as the weak light observation microscope of the first embodiment.

In the weak light observation microscope according to the fourth embodiment configured as described above, the weak light observation and the transmitted illumination observation using the normal light can be switched by the following two methods.
In the first method, the illumination light cut filter 1d is inserted between the objective lens 1a and the imaging lens 1b, and the light source 4a ′ of the illumination optical system 4 ′ is turned on. In this method, the illumination light cut filter 1d is inserted into and removed from the imaging lens 1b.
When faint light observation is performed, an illumination light cut filter 1d is inserted. Predetermined weak light emitted from the observation object 10 toward the light return optical system 2 side forms an image on the surface of the dichroic mirror 2c ″ through the second objective lens 2a and the second imaging lens 2b. , Reflected by the dichroic mirror 2c ", returns to the observation object 10 through the second imaging lens 2b and the second objective lens 2a. Further, the predetermined weak light transmitted through the observation target 10 toward the imaging optical system 1 side and the predetermined weak light emitted from the observation target 10 toward the imaging optical system 1 side pass through the objective lens 1a. Then, the light passes through the illumination light cut filter 1d, passes through the imaging lens 1b, and forms an image on the imaging surface of the imaging device 1c. Illumination light from the illumination optical system 4 ′ passes through the dichroic mirror 2c ″, passes through the second imaging lens 2b, the second objective lens 2a, the observation object 10, and the objective lens 1a, and then passes through the illumination light cut filter. 1d, but is blocked by the illumination light cut filter 1d.
On the other hand, when performing transmitted illumination observation with normal light, the illumination light cut filter 1d is extracted. The light from the illumination optical system 4 ′ passes through the dichroic mirror 2c ″, passes through the second imaging lens 2b, the second objective lens 2a, the observation object 10, the objective lens 1a, and the imaging lens 1b, and then passes through the imaging device. The image is formed on the imaging surface 1c.

The second method is a method of switching ON / OFF of the light source 4a ′ of the illumination optical system 4 ′ with the illumination light cut filter 1d extracted from between the objective lens 1a and the imaging lens 1b.
When performing faint light observation, the light source 4a ′ of the illumination optical system 4 ′ is turned off. Predetermined weak light emitted from the observation object 10 toward the light return optical system 2 side forms an image on the surface of the dichroic mirror 2c ″ through the second objective lens 2a and the second imaging lens 2b. , Reflected by the dichroic mirror 2c ", returns to the observation object 10 through the second imaging lens 2b and the second objective lens 2a. The predetermined weak light transmitted through the observation target 10 toward the imaging optical system 1 side and the predetermined weak light emitted from the observation target 10 toward the imaging optical system 1 side are connected to the objective lens 1a. An image is formed on the imaging surface of the imaging element 1c through the image lens 1b.
On the other hand, when performing transmitted illumination observation with normal light, the light source 4a ′ of the illumination optical system 4 ′ is turned ON. The illumination light from the illumination optical system 4 ′ is collected on the surface of the dichroic mirror 2c ″, transmitted through the dichroic mirror 2c ″, and passed through the second imaging lens and the second objective lens 2a to the observation object 10. The incident light that has passed through the observation object 10 forms an image on the imaging surface of the imaging element 1c via the objective lens 1a and the imaging lens 1b.

  According to the weak light observation microscope of the fourth embodiment, it is possible to switch between the weak light observation and the transmitted illumination observation with an extremely small number of members, and the same effects as the weak light observation microscope of the first embodiment can be obtained. That is, a predetermined weak light having a light quantity about twice that of a conventional weak light observation microscope is incident, and the exposure time of the image sensor 1c can be shortened to about half, and the low magnification objective lens that cannot increase the NA. Even if is used, it is possible to capture a light emission image, and it is possible to capture a fast change of the observation target (light emitting portion).

In the example of FIG. 4, the predetermined weak wavelength is described as the emission wavelength emitted from the observation target by a chemical reaction with the luminescent reagent or the like, but the predetermined weak wavelength is emitted from the observation target by irradiation with the excitation wavelength. It may be a fluorescence wavelength.
In that case, a band-pass filter having optical characteristics that shields a predetermined fluorescence wavelength and transmits a predetermined excitation light wavelength between the light source 4a ′ and the condenser lens 4b ′ may be detachably provided. .
In the fourth embodiment, the mirror disposed at the position conjugate with the observation object 10 in the light return optical system 2 is configured by the planar dichroic mirror 2c ″. However, the mirror is configured by a variable shape dichroic mirror. May be.
By doing so, by deforming the reflecting surface of the deformable dichroic mirror 2c ″ in accordance with the thickness or optical path length of the observation object 10, predetermined weak light reflected by the deformable dichroic mirror 2c ″ can be obtained. 10 can be accurately returned to the position of the light emitting point, and even if the thickness of the observation object 10 is not constant, blurring of the image can be prevented.

As mentioned above, although embodiment of the weak light observation microscope of this invention was described, the weak light observation microscope of this invention is not limited to these structures.
For example, it is preferable to provide an aberration correction means such as a correction ring or a phase modulator at the pupil position of the second objective lens 2a in each embodiment.
When the container 11 that accommodates the observation object 10 is a petri dish or a microplate, the liquid surface in the container 11 is bent by surface tension, and aberration may occur. If an aberration correction means is provided at the pupil position of the second objective lens 2a, it is possible to correct the aberration due to the bending of the liquid surface and form an image on the mirror 2c (2c ′, 2c ″).

  In each embodiment shown in FIGS. 1 to 4, if the mirrors 2 c, 2 c ′, 2 c ″ are arranged at a position conjugate with the observation target 10, the optical return optical system 2 is In order to make the image of weak light as bright as possible, it is preferable to use the second objective lens 2a having a larger NA than that of the objective lens 1a.

1 to 4, the light return optical system 2 is provided on the opposite side of the imaging optical system 1 with respect to the observation target 10, but for light return in the weak light observation microscope of the present invention. The optical system 2 is not limited to such an arrangement.
For example, the light return optical system 2 may be provided on the side of the observation target with respect to the observation target accommodated in the transparent cube-shaped container.

  The weak light observation microscope and the weak light acquisition method of the present invention are for observing an observation object that emits very weak light, such as long-time observation of changes in the fine structure level of cells and tissues due to light emission, fluorescence observation, etc. It is useful in all fields where is required.

DESCRIPTION OF SYMBOLS 1 Imaging optical system 1a Objective lens 1b Imaging lens 1c Image pick-up element 1d Illumination light cut filter 2 Optical return optical system 2a Second objective lens 2b Second imaging lens 2c (Plane) mirror 2c 'Variable shape mirror 2c Dichroic mirror 3 Optical path dividing member 4, 4 'Illumination optical system 4a, 4a' Light source 4b Collimator lens 4b 'Condensing lens 4c Bandpass filter 10 Observation object (sample)
10a Light emitting unit 10a ′, 10a ″ Image of light emitting unit 11 Container 51 Imaging optical system 51a Objective lens 51b Imaging lens 51c Imaging element 60 Observation target (sample)
60a Light emitting part 60a 'Image of light emitting part 61 Container

Claims (7)

A weak light observation microscope having an imaging optical system composed of an objective lens, an imaging lens, and an image sensor, and acquiring predetermined weak light emitted from an observation object via the imaging optical system;
The second objective lens, the second imaging lens, and the observation object are arranged at a conjugate position to reflect the wavelength of the predetermined weak light and transmit a wavelength other than the wavelength of the predetermined weak light. And a light return optical system having a mirror on the opposite side of the imaging optical system with respect to the observation target,
An illumination optical system is provided on the opposite side of the mirror from the second imaging lens and emits illumination light having a wavelength different from the wavelength of the predetermined weak light toward the mirror. A weak light observation microscope characterized by that.   2. The feeble light according to claim 1, wherein the imaging optical system further includes a wavelength selection element that shields the wavelength of the illumination light and allows the wavelength of the predetermined feeble light to pass through. Observation microscope.   The weak light observation microscope according to claim 1, wherein an aberration correction unit is provided at a pupil position of the second objective lens.   The weak light observation microscope according to claim 3, wherein the aberration correction unit is a correction ring.   The weak light observation microscope according to claim 3, wherein the aberration correction unit is a phase modulator.   The weak light observation microscope according to claim 1, wherein the predetermined weak light is luminescence emitted from the observation object by a chemical reaction. The predetermined weak light is fluorescence emitted from the observation object by irradiation with an excitation wavelength;
The weak light observation microscope according to claim 2, wherein the illumination light has an excitation wavelength that excites the observation object.

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