JP2008116756A - Optical device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an optical device by which the optical paths of scanning optical systems relating to light having a plurality of wavelengths can easily be combined without using a complicated configuration. <P>SOLUTION: The optical device includes: a stimulus light scanning optical system 17 for deflecting stimulus light that is used for stimulating a body 13 to be tested and has at least one or more wavelengths; and a combining acoustic optical element 33 for combining the optical path of the deflected stimulus light with the optical path of an observing optical system 15 that obtains the image of the body 13 being tested. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an optical device.

In recent years, functional analysis of living cells using a scanning confocal microscope or the like has been actively performed. By using this scanning confocal microscope or the like, functional analysis at various depths in a thick biological cell can be performed, and thus it has become possible to elucidate the three-dimensional structure of a biological cell.
In the scanning confocal microscope, a sample is irradiated with light emitted from a point light source, and fluorescence or reflected light emitted from the sample is detected by a photodetector to obtain sample information. A pinhole is arranged on the light incident surface side of the photodetector at a position conjugate with the observation surface, and the photodetector detects only the light passing through the pinhole and converts it into an electrical signal. . Therefore, the scanning confocal microscope can obtain an optical slice image at a desired position in the optical axis direction in a thick sample.
The scanning confocal microscope can obtain the three-dimensional information of the sample by sequentially acquiring the information of the sample while moving at least one of the sample and the objective lens in the optical axis direction (for example, Patent Document 1). reference.).
Japanese Patent Laid-Open No. 10-206742

Patent Document 1 described above includes a first scanning optical system and a second scanning optical system, and the first scanning optical system and the second scanning optical system are made to coincide with each other by using a dichroic mirror ( A scanning confocal microscope (laser scanning microscope) is disclosed.
With such a configuration, the first scanning optical system and the second scanning optical system can share one objective lens.

  However, in the configuration in which the optical paths of the first and second scanning optical systems are combined using a dichroic mirror, depending on the combination of the wavelengths of irradiation light used in each scanning optical system and the combination of wavelengths detected by the photodetector Therefore, it is necessary to change the wavelength characteristics of the dichroic mirror. Therefore, when changing the irradiation wavelength and the detection wavelength, the synthetic dichroic mirror must be replaced. In particular, as in the case of fluorescence observation of biological samples, if you want to vary the wavelength of the observation (fluorescence excitation) laser light, the stimulation laser light, and the detection wavelength depending on the type of specimen and the observation site, Since a dichroic mirror suitable for the combination of wavelengths has to be attached / detached or switched, there is a problem that the structure is complicated, the cost is increased, and the operation is complicated.

  The present invention has been made to solve the above-described problems, and provides an optical device that can easily cope with various combinations of wavelengths of light used for stimulation and observation without using a complicated configuration. For the purpose.

In order to achieve the above object, the present invention provides the following means.
The present invention relates to a scanning optical system for stimulating light that deflects stimulating light having a wavelength of at least one wavelength, and an observation optical system for obtaining an image of the subject. And an optical acousto-optic device for synthesizing the optical path of the deflected stimulation light.

  According to the present invention, since a synthetic acoustooptic device having a higher degree of freedom in wavelength of light that can be synthesized is used as compared with a dichroic mirror that synthesizes only light having a predetermined wavelength, a stimulus having at least one wavelength is used. The optical path of light is easily combined with the optical path of the observation optical system without using a complicated configuration. In other words, the acoustooptic device can cope with a change in the wavelength of the light beam to be synthesized (diffracted) by changing the frequency of the ultrasonic wave (elastic wave) excited by the crystal body. The optical path of stimulation light having one or more wavelengths can be easily combined with the optical path of the observation optical system.

  Furthermore, when using a plurality of the above dichroic mirrors, etc., it is necessary to adjust the arrangement angle of the plurality of dichroic mirrors, etc., whereas the optical device of the present invention determines the arrangement position of one synthetic acoustooptic element, etc. By simply adjusting, the optical path of the stimulus light having at least one wavelength can be synthesized with the optical path of the observation optical system.

  Note that the stimulating light that changes the specimen causes a photochemical reaction to the specimen (for example, fluorescent bleaching or release of the caged reagent), manipulation of the specimen, dissection, processing, etc. This is light that irradiates the subject with a purpose other than observation, such as performing.

In the above-described invention, it is desirable that the stimulation light scanning optical system and the synthetic acousto-optic element be insertable into and removable from the optical path of the observation optical system.
According to the present invention, stimulation light and observation optics that can be freely scanned and changed in position by attaching the above-described stimulation light scanning optical system and synthetic acoustic optical element to an optical apparatus that includes only the observation optical system. It is possible to easily realize an optical device that synthesizes the light from the system and focuses it on the subject.

In the above invention, it is desirable that the stimulation light scanning optical system is provided with a stimulation light deflecting unit for deflecting the stimulation light.
According to the present invention, the stimulation light is deflected by the stimulation light deflecting unit and collected on a predetermined region of the subject. For example, by providing a stimulation light deflecting unit that deflects the stimulation light of each wavelength, the stimulation light of each wavelength can be simultaneously focused on different predetermined regions.

In the above invention, the synthetic acoustooptic device deflects the stimulation light in one direction intersecting each other, and the stimulation light scanning optical system deflects the stimulation light in another direction intersecting each other. It is preferable that an acoustooptic element is provided, and the stimulation light deflected by the deflecting acoustooptic element and the other light beam are incident on the synthetic acoustooptic element.
According to the present invention, the stimulation light is deflected in one direction intersecting with each other by the synthetic acoustooptic element, and is deflected in the other direction intersecting with each other by the deflection acoustooptic element. Therefore, the stimulation light can be scanned in one and other directions intersecting each other.

In the above invention, it is desirable that the synthetic acousto-optic element diffracts the stimulation light and transmits light in the optical path of the observation optical system.
According to the present invention, since the optical path of the stimulating light is diffracted in the synthetic acoustooptic element, the optical path of the stimulating light bends in the synthetic acoustooptic element and is combined with the optical path of the light beam of the observation optical system that passes through the synthetic acoustooptic element Is done.

  In the above invention, it is preferable that an objective optical system for condensing the stimulation light on the subject is provided, and the synthetic acousto-optic element is disposed at a position conjugate with the pupil of the objective optical system.

  According to the present invention, when the stimulation light is deflected, the incident position of the stimulation light on the synthetic acousto-optic device does not vary, but only the incident angle of the stimulation light varies. Therefore, since only the incident angle of the stimulation light is changed as compared with the case where the synthetic acoustooptic element is not arranged at the conjugate position, the optical path of the stimulation light can be easily combined with the optical path of the light beam of the observation optical system. .

  In the above invention, it is preferable that an objective optical system for condensing the stimulation light on the subject is provided, and the synthetic acousto-optic element is disposed at a position conjugate with an image of the objective optical system.

According to the present invention, when the stimulation light is deflected, the incident angle of the stimulation light incident on the synthetic acousto-optic device does not vary, and the incident position only moves in parallel. Therefore, since the incident position of the stimulating light is simply moved in parallel as compared with the case where the synthetic acoustooptic element is not arranged at the conjugate position, the optical path of the stimulating light can be easily combined with the optical path of the light beam of the observation optical system. .
For example, an acoustooptic device such as an acoustic modulation filter has a narrow allowable range of the incident angle of light. Therefore, the incident angle of the stimulation light does not change compared to the case where the synthetic acousto-optic element is arranged at a position conjugate with the pupil of the objective optical system, so the optical path of the stimulation light is changed to the optical path of the light beam of the observation optical system. Easy to synthesize.

  In the above-described invention, a signal generation unit that generates a modulation signal for controlling an ultrasonic wave excited in the synthetic acoustooptic device is provided, and the signal generation unit controls the ultrasonic wave, thereby It is desirable to control the intensity of the deflected stimulation light that is irradiated.

According to the present invention, the ultrasonic wave excited by the synthetic acoustooptic device is controlled by the signal control unit, and the intensity of the stimulation light irradiated on the subject can be freely controlled.
Here, the synthetic acoustooptic device diffracts part of the incident light beam and transmits the remaining light beam. The ratio of the diffracted light beam to the transmitted light beam depends on the amplitude of the ultrasonic wave excited by the synthetic acoustooptic device. In other words, the ratio of the diffracted light beam increases as the amplitude of the ultrasonic wave increases, and conversely, the ratio of the diffracted light beam decreases as the amplitude of the ultrasonic wave decreases. Therefore, the signal control unit inputs the modulation signal to the synthetic acousto-optic element, thereby controlling the amplitude of the ultrasonic wave and the amplitude, and freely controlling the intensity of the stimulation light irradiated to the subject. Can do.

  In the above invention, a detection unit that detects the intensity of the deflected stimulation light that passes through the synthetic acoustooptic device without being irradiated on the subject is provided, and the signal generation unit is detected by the detection unit. Preferably, the modulation signal is generated based on the intensity of the stimulated light.

According to the present invention, the signal generation unit generates a control signal based on the intensity of the stimulation light transmitted through the synthetic acoustooptic element detected by the detection unit, and inputs the control signal to the synthetic acoustooptic element, thereby irradiating the subject. The intensity of the stimulated light can be freely controlled.
Here, a part of the stimulation light incident on the synthetic acoustooptic element is diffracted by the synthetic acoustooptic element, and the remaining stimulation light is transmitted through the synthetic acoustooptic element. The signal generation unit performs control based on the intensity of the remaining stimulation light transmitted through the synthetic acoustooptic element detected by the detection unit, and thereby the intensity of the stimulation light diffracted by the synthesis acoustooptic element and the transmitted stimulation light. The strength of the can be controlled freely.

In the above invention, it is desirable that the synthetic acoustooptic element is an acoustooptic modulation filter.
According to the present invention, since an acousto-optic modulation filter (hereinafter referred to as AOTF) is used as a synthetic acousto-optic device, the AOTF diffracts the stimulation light and the optical path of the stimulation light of the observation optical system. It can be combined with the optical path of the light beam.

In the above invention, it is desirable that the synthetic acoustooptic element is an acoustooptic modulator.
According to the present invention, since an acousto-optic modulator (hereinafter referred to as AOM) is used as a synthetic acousto-optic device, the AOM diffracts the stimulation light, and the optical path of the stimulation light is the light beam of the observation optical system. Can be combined in the optical path.

In the above invention, it is desirable that the synthetic acoustooptic element is an acoustooptic deflector.
According to the present invention, since an acousto-optic deflector (hereinafter referred to as “AOD”) is used as a synthetic acousto-optic device, the AOD diffracts the stimulation light, and the optical path of the stimulation light is a light beam of the observation optical system. Can be combined in the optical path.

In the above invention, in any of the above inventions, it is desirable that the observation optical system includes a scanning optical system that scans light that irradiates the subject.
According to the present invention, a scan image of a subject can be acquired. For example, a confocal image is acquired using a confocal detection system, or a scan image is acquired using a multiphoton excitation method. Thus, an optical slice image of the sample can be acquired as an observation image.

  In addition, the objective light that combines the stimulus light deflected by the scanning optical system for stimulus light and the light beam deflected by the scanning optical system for observation is synthesized by a synthetic acousto-optic device, for example, condenses the light on the subject. Since the light is incident on the system, the stimulus light and the other light are deflected independently within the field of view of the same objective optical system. That is, the stimulation light and the light beam of the observation optical system are condensed on different predetermined areas in the subject.

In the above invention, in any one of the above inventions, it is desirable that the observation optical system is a non-scanning microscope optical system.
According to the present invention, a microscopic image of a subject can be obtained.

  According to the optical device of the present invention, since the stimulus light deflected by the stimulus light scanning optical system having at least one wavelength and the light beam of the observation optical system are incident on the synthetic acousto-optic element, a complicated There is an effect that it is possible to easily cope with various combinations of wavelengths of light used for stimulation and observation without using a configuration.

[First Embodiment]
The laser scanning microscope according to the first embodiment of the present invention will be described below with reference to FIGS.
FIG. 1 is a schematic diagram illustrating a configuration of a laser scanning microscope according to the present embodiment.
As shown in FIG. 1, the laser scanning microscope 1 includes a first light source 3, a first scanning optical system (observation optical system) 15, an optical device 7, a second light source 5, and a detection optical system 9. And an objective lens (objective optical system) 55 and a third relay optical system 53.

  The first light source 3 emits laser light used for obtaining an image of the subject 13. Specifically, the first light source 3 emits excitation light (light of the observation optical system) that is irradiated on the subject 13 to excite fluorescence. The excitation light emitted from the first light source 3 is incident on a first scanning optical system 15 described later.

  The first scanning optical system 15 deflects the excitation light and causes the deflected excitation light to enter the objective lens 55. The excitation light is deflected by the first scanning optical system 15 to scan the subject 13. The first scanning optical system 15 includes a dichroic mirror 19, a first scanning optical unit (scanning optical system) 21, and a first relay lens system 23.

  The dichroic mirror 19 transmits the excitation light and reflects the fluorescence excited by the subject 13. The dichroic mirror 19 is arranged between the first light source 3 and the first scanning optical unit 21 and arranged so that the fluorescence reflected by the dichroic mirror 19 enters the detection optical system 9. .

  The first scanning optical unit 21 scans the excitation light on the subject 13 by deflecting the excitation light two-dimensionally. The first scanning optical unit 21 is disposed between the dichroic mirror 19 and the first relay lens system 23. The first scanning optical unit 21 includes two first galvanometer mirrors 21A and 21B. The first galvanometer mirrors 21A and 21B deflect the excitation light in one and other directions orthogonal to each other.

The first relay lens system 23 cooperates with the third relay lens system 53 to relay the pupil of the objective lens 55 to the position of the first scanning optical unit 21. The first relay lens system 23 is disposed between the first scanning optical unit 21 and the objective lens 55.
Further, a mirror 27 is disposed between the first relay lens system 23 and the objective lens 55. The mirror 27 reflects the excitation light emitted from the first relay lens system 23 toward the objective lens 55.

  The optical device 7 irradiates stimulation light to a desired point or region of the specimen. The optical device 7 includes a second scanning optical system (stimulating light scanning optical system) 17 into which the stimulation light from the second light source 5 is incident, and a beam combining device 16.

  The second light source 5 emits laser light that is incident on a predetermined region of the subject 13. For example, the second light source 5 emits stimulation light that stimulates a predetermined region of the subject 13. By irradiating the stimulation light from the second light source 5 in this way, a predetermined reaction can be caused in the subject 13 and the reaction can be observed. Stimulation light emitted from the second light source 5 is incident on a second scanning optical system 17 of the optical device 7 to be described later.

  The second scanning optical system 17 deflects the stimulation light from the second light source 5. Stimulation light is deflected by the second scanning optical system 17 to irradiate a predetermined region of the subject 13. The second scanning optical system 17 includes a second scanning optical unit (stimulating light deflecting unit) 29 and a second relay lens system 31.

  The second scanning optical unit 29 irradiates the predetermined region of the subject 13 with the stimulation light by deflecting the stimulation light two-dimensionally. The second scanning optical unit 29 is disposed between the second light source 5 and the second relay lens system 31. The second scanning optical unit 29 includes two second galvanometer mirrors 29A and 29B. The second galvanometer mirrors 29A and 29B deflect the excitation light in one and other directions orthogonal to each other.

  The beam synthesis device 16 includes an AOTF (synthetic acoustooptic device) 33 and a control unit 43. Since the AOTF 33 diffracts only the wavelength of the stimulation light from the second scanning optical system 17 toward the objective lens 55, the observation laser light from the first scanning optical system and the observation light from the specimen differing from the wavelength of the stimulation light Is transmitted as it is.

  The second relay lens system 31 relays the pupil of the objective lens 55 to the position of the second scanning optical unit 29 in cooperation with the third relay optical system 53. The second relay lens system 31 is disposed between the second scanning optical unit 29 and the AOTF 33.

FIG. 2 is a schematic diagram for explaining a configuration in the vicinity of the optical device in FIG.
The AOTF 33 combines the optical path of the first scanning optical system 15 and the optical path of the second scanning optical system 17. Specifically, the AOTF 33 synthesizes the optical path of the excitation light and the optical path of the stimulation light by transmitting the excitation light and diffracting the stimulation light, and enters the objective lens 55.

  The AOTF 33 is disposed between the second relay lens system 31 and the third relay lens system 53, and the AOTF 33 is disposed between the first relay lens system 23 and the third relay lens system 53. Yes. As shown in FIG. 2, the AOTF 33 includes a transducer 37, a crystal body 39, and an ultrasonic absorber 41.

The transducer 37 is for exciting the crystal body 39 with ultrasonic waves. The transducer 37 is disposed in contact with the crystal body 39, and a high frequency voltage is applied from the control unit 43.
The crystal body 39 diffracts incident stimulus light by excited ultrasonic waves. A transducer 37 is disposed on one surface of the crystal body 39, and an ultrasonic absorber 41 is disposed on the other surface opposite to the one surface. The ultrasonic absorber 41 absorbs the ultrasonic wave that is excited by the transducer 37 and propagates through the crystal body 39.

The AOTF 33 is connected to a control unit (signal generation unit) 43 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOTF 33.
The control unit 43 includes a signal generation unit 43A that generates a modulation signal for controlling ultrasonic waves, and a high-frequency transmitter 43B that generates a high-frequency voltage based on the modulation signal.

The high frequency transmitter 43B of the control unit 43 generates a high frequency voltage that excites an ultrasonic wave that diffracts the stimulation light. The signal generation unit 43A of the control unit 43 generates a modulation signal that causes the high-frequency transmitter 43B to excite the ultrasonic waves.
Thus, the control unit 43 can control the wavelength and intensity of the stimulation light diffracted toward the objective lens 55 by controlling the frequency and amplitude of the ultrasonic wave (elastic wave) excited in the AOTF 33. it can.

  As the AOTF 33, a known acousto-optic modulation filter can be used and is not particularly limited.

  The detection optical system 9 detects fluorescence excited by the subject 13. As shown in FIG. 1, the detection optical system 9 is a confocal detection system including a photometric filter 45, a lens 47, a pinhole 49, and a photoelectric conversion element 51.

The photometric filter 45 is a filter that transmits only the fluorescence excited from the subject 13. The photometric filter 45 is disposed between the dichroic mirror 19 and the lens 47.
The lens 47 forms an image of the fluorescence that has passed through the photometric filter 45 on the surface of the pinhole 49. The lens 47 is disposed between the photometric filter 45 and the pinhole 49.

The pinhole 49 is a through-hole through which only the fluorescence excited from a predetermined observation cross section of the subject 13 passes. The pinhole 49 is disposed at a position conjugate with the focal position of the objective lens 55.
The photoelectric conversion element 51 converts the intensity of incident fluorescence into an electric signal. The photoelectric conversion element 51 is disposed at a position where the fluorescence that has passed through the pinhole 49 enters.

The objective lens 55 and the third relay optical system 53 condense the excitation light and stimulation light emitted from the first scanning optical system 15 and the second scanning optical system 17 toward the subject 13.
The objective lens system 55 is disposed between the optical device 7 and the subject 13. The third relay lens system 53 relays light from the sample collected by the objective lens 55. The objective lens 55 collects excitation light and stimulation light on the cross section 57 of the subject 13. In addition, as shown in FIG. 2, you may arrange | position a reflective mirror between the objective lens 55 and AOTF33, It does not specifically limit.

Next, the operation of the laser scanning microscope 1 having the above configuration will be described.
As shown in FIG. 1, the excitation light emitted from the first light source 3 passes through the dichroic mirror 19 and then enters the first scanning optical unit 21. The first scanning optical unit 21 deflects the excitation light in a predetermined direction by the first galvanometer mirrors 21A and 21B.

Specifically, the first galvanometer mirror 21A deflects the excitation light incident from the first light source 3 in one direction orthogonal to each other. The first galvanometer mirror 21B deflects the excitation light deflected by the first galvanometer mirror 21A in other directions orthogonal to each other.
The excitation light deflected by the first scanning optical unit 21 passes through the first relay lens system 23 and then enters the mirror 27. The mirror 27 reflects the incident excitation light toward the AOTF 33. The excitation light is incident on the AOTF 33 from the 0th-order diffraction direction of the AOTF 33.
In this way, the excitation light is deflected by the first scanning optical unit 21 and thereby condensed at a desired position of the subject 13.

On the other hand, the stimulus light emitted from the second light source 5 is incident on the second scanning optical unit 29. The second scanning optical unit 29 deflects the stimulation light in a predetermined direction by the second galvanometer mirrors 29A and 29B.
Specifically, the second galvanometer mirror 29A deflects the stimulation light incident from the second light source 5 in one direction orthogonal to each other. The second galvanometer mirror 29B deflects the excitation light deflected by the second galvanometer mirror 29A in other directions orthogonal to each other.

The deflected stimulation light passes through the second relay lens system 31 and then enters the AOTF 33. Stimulation light is incident on the AOTF 33 from the first-order diffraction direction of the AOTF 33.
In this way, the stimulation light is deflected by the second scanning optical unit 29 and is collected at a desired position of the subject 13.

As described above, the deflected excitation light and the deflected stimulation light are incident on the AOTF 33 in the manner shown in FIG.
A high frequency voltage is applied from the control unit 43 to the transducer 37 of the AOTF 33. The transducer 37 excites the ultrasonic wave in the crystal body 39. The excited ultrasonic wave propagates through the crystal body 39 from the transducer 37 side toward the ultrasonic absorber 41 side.

The excitation light incident in the 0th order diffraction direction of the AOTF 33 from the first scanning optical system 15 passes through the AOTF 33 and enters the third relay optical system 53.
On the other hand, the stimulation light incident in the first-order diffraction direction of the AOTF 33 from the second scanning optical system 17 is diffracted by the AOTF 33 and enters the third relay optical system 53 together with the excitation light. Specifically, the stimulation light is diffracted by a periodic structure having a refractive index formed on the crystal body 39 by ultrasonic waves.
In this way, the optical path of the stimulation light is synthesized with the optical path of the excitation light that passes through the AOTF 33 by being diffracted and bent by the AOTF 33.

The control unit 43 controls the intensity of the stimulation light incident on the subject 13 by controlling the voltage amplitude of the high frequency voltage.
That is, the control unit 43 generates a modulation signal that changes the voltage amplitude of the high-frequency voltage from the internal signal generation unit 43A, and the high-frequency transmitter 43B generates a high-frequency voltage based on the modulation signal. The generated high frequency voltage is input to the transducer 37 of the AOTF 33, and an ultrasonic wave having an amplitude corresponding to the voltage amplitude of the high frequency voltage is excited. When the amplitude of the ultrasonic wave changes, the refractive index in the periodic structure of the refractive index formed in the crystal body 39 changes. Since the ratio of the stimulation light diffracted by the AOTF 33 and the stimulation light transmitted through the AOTF 33 changes with the change in the refractive index, the control unit 43 can control the intensity of the stimulation light incident on the subject 13.

Excitation light and stimulation light incident on the third relay optical system 53 are incident on the objective lens 55 and are respectively collected on the cross section 57 of the subject 13.
Since the excitation light and the stimulation light collected on the cross section 57 are deflected by the first scanning optical unit 21 and the second scanning optical unit 29, respectively, they are scanned separately. For example, the excitation light is scanned so as to irradiate the entire surface of the observation region in order to obtain an image of the cross section 57, while the stimulation light is scanned so as to irradiate only a predetermined region in the cross section 57.

Excitation light reflected from the subject 13 and fluorescence excited from the subject 13 by irradiation with the excitation light (hereinafter referred to as fluorescence) propagates in the opposite direction on the optical path through which the excitation light has propagated. Then, the light enters the detection optical system 9.
Specifically, fluorescence or the like first enters the objective lens 55 and then enters the AOTF 33 from the third relay optical system 53. The fluorescence or the like incident on the AOTF 33 passes through the AOTF 33 and enters the first scanning optical system 15. Note that the reflected stimulus light included in the fluorescence or the like is diffracted by the AOTF 33 and thus does not enter the first scanning optical system 15.

  The fluorescence or the like incident on the first scanning optical system 15 enters the dichroic mirror 19 via the mirror 27, the first relay lens system 23, and the first scanning optical unit 21. The dichroic mirror 19 reflects only the fluorescence excited from the subject 13 and transmits the excitation light reflected from the subject 13.

The fluorescence reflected by the dichroic mirror 19 enters the detection optical system 9. The fluorescence that has entered the detection optical system 9 enters the photometric filter 45. The photometric filter 45 transmits only desired fluorescence from the subject 13 while shielding excitation light and the like incident on the detection optical system 9.
The fluorescence transmitted through the photometric filter 45 enters the lens 47 and forms an image on the pinhole 49. The pinhole 49 passes only the fluorescence excited from the cross section 57 of the subject 13 among the fluorescence excited from the subject 13. The fluorescence that has passed through the pinhole 49 is incident on the photoelectric conversion element 51. The photoelectric conversion element 51 detects the intensity of incident fluorescence and outputs a detection signal corresponding to the intensity.

  The laser scanning microscope 1 detects fluorescence in the cross section 57 of the subject 13 based on the detection signal output from the photoelectric conversion element 51 and the control information of the first galvanometer mirrors 21A and 21B of the first scanning optical unit 21. Images can be acquired. Furthermore, the laser scanning microscope 1 can acquire a three-dimensional fluorescence image of the subject 13 by acquiring the fluorescence image while changing the position of the cross section 57 relative to the subject 13 in the optical axis direction.

According to the above configuration, since the AOTF 33 having a higher degree of freedom of wavelength of light that can be synthesized is used as compared with a dichroic mirror that synthesizes only light having a predetermined wavelength, the stimulation light having at least one wavelength is used. The optical path can be easily combined with the optical path of the excitation light without using a complicated configuration.
That is, since the AOTF 33 can cope with a change in the wavelength of the light beam to be synthesized (diffracted) by changing the frequency of the ultrasonic wave (elastic wave) excited by the crystal 39, the optical device 7 of the present embodiment has at least The optical path of stimulation light having one or more wavelengths can be easily combined with the optical path of excitation light.

  Furthermore, the optical system described in Patent Document 1 that uses a plurality of the dichroic mirrors and the like needs to be adjusted with respect to the arrangement angle and the like of the plurality of dichroic mirrors and the like. Even if the wavelength of the stimulation light is changed, the optical path of the stimulation light can be synthesized with the optical path of the excitation light only by adjusting the arrangement position of one AOTF 33 or the like.

  Further, since the stimulation light deflected by the second scanning optical system 17 and the excitation light deflected by the first scanning optical system 15 are combined by the AOTF 33 and incident on the objective lens 55, the same objective lens is used. Stimulation light and excitation light can be deflected independently within 55 fields of view. That is, the stimulation light and the excitation light can be condensed on different predetermined regions in the subject 13.

  Since the AOTF 33 can cope with a change in the wavelength of the light beam to be synthesized (diffracted) by changing the frequency of the ultrasonic wave (elastic wave) excited by the crystal body 39, the AOTF 33 can be combined with the first scanning optical system 15 with one AOTF 33. Regardless of the wavelength of light used in each of the second scanning optical systems 17, both optical paths can be easily combined. As a result, compared with the optical system described in Patent Document 1 in which a dichroic mirror corresponding to the wavelength of the stimulation light used in the second scanning optical system 17 needs to be arranged, the AOTF 33 has one stimulation light having a different wavelength. Therefore, the structure of the optical device 7 can be simplified.

  As in the above-described embodiment, an AOTF 33, an acoustooptic modulator (AOM), or an acoustooptic deflector (AOD) may be used as an acoustooptic element. It is not limited. This is because the optical paths of the first scanning optical system 15 and the second scanning optical system 17 can be combined using any acousto-optic element.

  Note that, as in the above-described embodiment, the stimulation light should be stimulated not only by scanning the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B of the second scanning optical unit 29. If the region is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to one minute point.

[First Modification of First Embodiment]
Next, a first modification of the first embodiment of the present invention will be described with reference to FIG.
The basic configuration of the laser scanning microscope of this modification is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIG. 3, and description of other components and the like will be omitted.
FIG. 3 is a schematic diagram for explaining the configuration in the vicinity of the optical device of the laser scanning microscope in the present modification.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

As shown in FIG. 3, the laser scanning microscope 101 includes a first scanning optical system 15 on which excitation light is incident, an optical device 107, and an objective lens 55.
The optical device 107 deflects the incident stimulus light and combines the deflected stimulus light optical path and the excitation light optical path. The optical device 107 includes a second scanning optical system (stimulating light scanning optical system) 117 into which stimulation light having three different wavelengths is incident, and an AOFT 33.

The second scanning optical system 117 deflects incident stimulation light having three different wavelengths.
The stimulation light incident on the second scanning optical system 117 is the first stimulation light, the second stimulation light, and the third stimulation light, each having a different wavelength. Stimulation light is deflected by the second scanning optical system 117 to irradiate a predetermined region of the subject 13.

  The second scanning optical system 117 includes a first dichroic mirror 118A, a second dichroic mirror 118B, a third mirror 118C, a second scanning optical unit 29, and an AOTF 33.

The first dichroic mirror 118A transmits the first stimulation light, and reflects the second stimulation light and the third stimulation light, and includes the optical path of the first stimulation light, the second and second stimulation lights, 3 and the optical path of the stimulating light.
The second dichroic mirror 118B reflects the second stimulation light and transmits the third stimulation light, and combines the optical path of the second stimulation light and the optical path of the third stimulation light. To do.
The third mirror 118C reflects the third stimulus light.

  The second scanning optical unit 29 is configured to cause each stimulation light to enter a predetermined region of the subject 13 by deflecting three different wavelengths of the stimulation light combined in the optical path. The second scanning optical unit 29 is disposed between the first dichroic mirror 118A and the AOTF 33.

A control unit (signal generation unit) 143 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOTF 33 is disposed in the AOTF 33. The control unit 143 includes a signal generation unit 143A that generates a modulation signal that controls ultrasonic waves, and a high-frequency transmitter 143B that generates a high-frequency voltage based on the modulation signal. The high-frequency transmitter 143B of the control unit 143 generates high-frequency voltages of three different frequencies that excite ultrasonic waves that diffract each stimulus light.
Further, the signal generation unit 143A of the control unit 43 generates three different modulation signals that cause the high-frequency transmitter 143B to excite the ultrasonic waves.

Next, the operation of the laser scanning microscope 101 having the above configuration will be described.
Action until the excitation light is emitted from the first light source 3 (see FIG. 1) and applied to the subject 13, and fluorescence excited from the subject 13 is detected by the photoelectric conversion element 51 (see FIG. 1). Since the operation up to this point is the same as that of the first embodiment, the description thereof is omitted.

As shown in FIG. 3, the first stimulation light, the second stimulation light, and the third stimulation light are incident on the first dichroic mirror 118A, the second dichroic mirror 118B, and the third mirror 118C, respectively. .
The first stimulation light passes through the first dichroic mirror 118A and enters the AOTF 33. The second stimulation light is reflected by the second dichroic mirror 118B toward the first dichroic mirror 118A, and further reflected by the first dichroic mirror 118A toward the AOTF 33. The third stimulation light is reflected toward the second dichroic mirror 118B by the third mirror 118C. The reflected third stimulation light is transmitted through the second dichroic mirror 118B and reflected toward the AOTF 33 by the first dichroic mirror 118A.

The first stimulation light, the second stimulation light, and the third stimulation light, which are optical path synthesized, are incident on the AOTF 33. Each stimulus light is diffracted by the AOTF 33 in accordance with the high-frequency voltage signal from the control unit 143 and is incident on the objective lens 55 together with the excitation light.
The control unit 143 inputs high frequency voltages having three types of frequencies corresponding to the first stimulation light, the second stimulation light, and the third stimulation light to the transducer 37 of the AOTF 33. The transducer 37 excites an ultrasonic wave corresponding to each frequency of the high frequency voltage to the crystal body 39. The crystal body 39 is formed with a periodic structure having a refractive index having a period corresponding to each stimulation light, and each stimulation light is diffracted by the AOTF 33 and incident on the objective lens 55 together with the excitation light.

In addition, by appropriately changing the high-frequency voltage input from the control unit 143 to the AOTF 33, it is possible to freely select which stimulation light of the three types of stimulation light is irradiated to the subject 13 (specimen). Can be adjusted. For example, when it is desired to irradiate the subject 13 with only the first stimulus light, only the high-frequency voltage having a frequency corresponding to the wavelength of the first stimulus light may be input to the AOTF 33, and the first and second stimulus lights may be input. When it is desired to irradiate, a high frequency voltage corresponding to the wavelength of each of the first and second stimulation lights may be input.
Furthermore, when irradiating a plurality of types of stimulating light, the intensity (amplitude) of the high-frequency voltage corresponding to each wavelength is appropriately adjusted, so that the intensity of the stimulating light irradiated to the subject 13 can be freely adjusted. .
Since the subsequent operation is the same as that of the first embodiment, the description thereof is omitted.

According to the above configuration, since the AOTF 33 having a higher degree of freedom in wavelength of light that can be synthesized is used compared to a dichroic mirror that synthesizes only light having a predetermined wavelength, the first stimulation light and the second stimulation light are used. The optical paths of the light and the third stimulation light can be easily combined with the optical path of the excitation light without using a complicated configuration.
In other words, the AOTF 33 can cope with a change in the wavelength of the light beam to be synthesized (diffracted) by changing the frequency of the ultrasonic wave (elastic wave) excited by the crystal body 39. The optical paths of the first stimulation light, the second stimulation light, and the third stimulation light can be easily combined with the optical path of the excitation light.

  Furthermore, the optical system described in Patent Document 1 that uses a plurality of dichroic mirrors or the like needs to be adjusted with respect to the arrangement angle or the like of the plurality of dichroic mirrors or the like, whereas the optical device 107 according to this modification of the present embodiment. Can synthesize the optical path of the stimulation light having at least one wavelength with the optical path of the excitation light only by adjusting the arrangement position or the like of one AOTF 33.

  In addition, since the stimulation light deflected by the second scanning optical system 107 and the excitation light deflected by the first scanning optical system 15 are incident on the AOTF 33, the stimulation light and the excitation light are independently generated. Can be deflected. That is, the first stimulation light, the second stimulation light, the third stimulation light, and the excitation light can be condensed on different predetermined areas in the subject 13.

  Note that, as in the above-described modification, the stimulation light is only scanned in the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. Instead, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to the minute point.

[Second Embodiment]
Next, a second embodiment of the present invention will be described with reference to FIGS.
The basic configuration of the laser scanning microscope of this embodiment is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIGS. 4 and 5, and description of other components will be omitted.
FIG. 4 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope in the present embodiment.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

  As shown in FIG. 4, the laser scanning microscope 201 includes a first scanning optical system (observation optical system) 215 that receives excitation light, an optical device 207, a third relay lens system 219, and an objective lens 55. And.

  The first scanning optical system 215 deflects the incident excitation light and makes the deflected excitation light enter the objective lens 55. The excitation light is deflected by the first scanning optical system 215 to scan the subject 13. The first scanning optical system 215 includes a dichroic mirror 19 (see FIG. 1), a first scanning optical unit 21, and a first relay lens system 223.

  The first scanning optical unit 21 scans the excitation light on the subject 13 by deflecting the excitation light. The first scanning optical unit 21 is disposed between the dichroic mirror 19 and the first relay lens system 223 and at a position 21R conjugate with the pupil 11R of the objective lens 55.

  The first relay lens system 223 causes the deflected excitation light to enter the AOTF 33. The first relay lens system 223 is arranged between the first scanning optical unit 21 and the third relay lens system 219, and the arrangement position 21R of the first scanning optical unit 21 is the pupil 11R of the objective lens 55. It is arranged so as to be a conjugate position.

  The optical device 207 deflects incident excitation light and stimulation light, respectively. The optical device 207 includes a second scanning optical system (stimulating light scanning optical system) 217 on which stimulation light is incident, and an AOTF 33.

  The second scanning optical system 217 deflects the incident stimulation light and combines the deflected stimulation light optical path and the deflected excitation light optical path. Stimulation light is deflected by the second scanning optical system 217 to irradiate a predetermined region of the subject 13. The second scanning optical system 217 includes a second scanning optical unit 29 and a second relay lens system 231.

  The second scanning optical unit 29 is for making the stimulation light incident on a predetermined region of the subject 13 by deflecting the stimulation light. The second scanning optical unit 29 is disposed between the second light source 5 (see FIG. 1) and the second relay lens system 231 and at a position 29R conjugate with the pupil 11R of the objective lens 55. .

  The second relay lens system 231 makes the deflected excitation light incident on the AOTF 33. The second relay lens system 231 is arranged between the second scanning optical unit 29 and the AOTF 33 so that the arrangement position of the second scanning optical unit 29 is a conjugate position 29R with the pupil 11R of the objective lens 55. Is arranged.

  The AOTF 33 combines the optical path of the first scanning optical system 215 and the optical path of the second scanning optical system 217. The AOTF 33 is disposed at a position 33R conjugate with the pupil 11R of the objective lens 55.

The third relay lens system 219 corresponds to the third relay optical system 53 in FIG. 1 and relays the pupil 11R of the objective lens 55 to the AOTF 33.
The third relay lens system 219 is arranged between the AOTF 33 and the objective lens 55, and relays the pupil 11R so that the arrangement position of the AOTF 33 becomes a position 33R conjugate with the pupil 11R of the objective lens 55.

Next, the operation of the laser scanning microscope 201 having the above configuration will be described.
The description of the same parts as those in the first embodiment is omitted.

FIG. 5 is a schematic diagram showing the state of light rays in the optical device of FIG.
As shown in FIG. 5, the excitation light incident on the first scanning optical system 215 is deflected by the first scanning optical unit 21 disposed at a position 21R conjugate with the pupil 11R of the objective lens 55 to be first. Is incident on the relay lens system 223. The excitation light incident on the first relay lens system 223 is incident on the AOTF 33 disposed at a position 33R conjugate with the pupil 11R of the objective lens 55.

The deflected excitation light is incident on the same region of the AOTF 33 as compared with that before the deflection, and is incident on the AOTF 33 at a different incident angle.
The excitation light incident on the AOTF 33 enters the objective lens 55 and is collected on the subject 13.

  On the other hand, the stimulating light incident on the second scanning optical system 217 is deflected by the second scanning optical unit 29 disposed at a position 29R conjugate with the pupil 11R of the objective lens 55, and the second relay lens system 231. Is incident on. The stimulus light incident on the second relay lens system 231 is incident on the AOTF 33 disposed at a position 33R conjugate with the pupil 11R of the objective lens 55.

The deflected stimulation light is incident on the same region of the AOTF 33 as compared with that before the deflection, and is incident on the AOTF 33 at a different incident angle.
The stimulation light incident on the AOTF 33 is diffracted, incident on the objective lens 55 together with the excitation light, and collected on the subject 13.

  According to the above configuration, when the stimulation light is deflected, the incident position of the stimulation light on the AOTF 33 does not vary, and only the incident angle of the stimulation light varies. Therefore, since only the incident angle of the stimulating light is changed as compared with the case where the AOTF 33 is not disposed at the position 33R conjugate with the pupil 11R of the objective lens 55, the optical path of the stimulating light is easily combined with the optical path of the excitation light. it can.

  Note that, as in the above-described embodiment, the stimulation light is only scanned in the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. Instead, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to the minute point.

[Third Embodiment]
Next, a third embodiment of the present invention will be described with reference to FIGS.
The basic configuration of the laser scanning microscope of this embodiment is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIGS. 6 and 7, and description of other components will be omitted.
FIG. 6 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope according to the present embodiment.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

  As shown in FIG. 6, the laser scanning microscope 301 includes a first scanning optical system (observation optical system) 315 to which excitation light is incident, an optical device 307, a third relay lens system 319, and an objective lens 55. And.

  The first scanning optical system 315 deflects the incident excitation light and causes the deflected excitation light to enter the objective lens 55. The excitation light is deflected by the first scanning optical system 315 to scan the subject 13. The first scanning optical system 315 includes a dichroic mirror 19 (see FIG. 1), a first scanning optical unit 21, and a first relay lens system 323.

  The first scanning optical unit 21 scans the excitation light on the subject 13 by deflecting the excitation light. The first scanning optical unit 21 is disposed between the dichroic mirror 19 and the first relay lens system 323 at a position 21R conjugate with the pupil 11R of the objective lens 55.

  The first relay lens system 323 causes the deflected excitation light to enter the AOTF 33. The first relay lens system 323 is arranged between the first scanning optical unit 21 and the third relay lens system 319, and the arrangement position 21R of the first scanning optical unit 21 is the pupil 11R of the objective lens 55. It is arranged so as to be a conjugate position.

  The optical device 307 deflects the stimulation light and introduces it into the optical path of the objective lens 55. The optical device 307 includes a second scanning optical system (stimulating light scanning optical system) 317 on which stimulation light is incident, and an AOTF 33.

  The second scanning optical system 317 deflects the incident stimulation light and combines the deflected stimulation light optical path and the deflected excitation light optical path. The stimulation light is deflected by the second scanning optical system 317, and is irradiated on a predetermined region of the subject 13. The second scanning optical system 217 includes a second scanning optical unit 29 and a second relay lens system 331.

  The second scanning optical unit 29 is for making the stimulation light incident on a predetermined region of the subject 13 by deflecting the stimulation light. The second scanning optical unit 29 is disposed between the second light source 5 and the second relay lens system 331 and at a position 29R conjugate with the pupil 11R of the objective lens 55.

  The second relay lens system 331 allows the deflected excitation light to enter the AOTF 33. The second relay lens system 331 is arranged between the second scanning optical unit 29 and the AOTF 33 so that the arrangement position of the second scanning optical unit 29 is a position 29R conjugate with the pupil 11R of the objective lens 55. Is arranged.

  The AOTF 33 combines the optical path of the first scanning optical system 315 and the optical path of the second scanning optical system 17. The AOTF 33 is disposed at a position 33M conjugate with the focal position (observation cross section) of the objective lens 55.

The third relay lens system 319 corresponds to the third relay optical system 53 in FIG. 1 and forms an image on the AOTF 33 with light from the sample collected by the objective lens 55.
The third relay lens system 319 makes deflected excitation light and stimulation light enter the objective lens 55. The third relay lens system 319 is arranged between the AOTF 33 and the objective lens 55, and forms an image of light from the objective lens 55 so that the arrangement position of the AOTF 33 is a position 33M conjugate with the focal position of the objective lens 55. Let

Next, the operation of the laser scanning microscope 301 having the above configuration will be described.
The description of the same parts as those in the first embodiment is omitted.

FIG. 7 is a schematic diagram showing the state of light rays in the optical device of FIG.
The excitation light incident on the first scanning optical system 315 is deflected by the first scanning optical unit 21 and incident on the first relay lens system 323 as shown in FIG. The excitation light incident on the first relay lens system 323 is incident on the AOTF 33 disposed at a position 33M conjugate with the focal position of the objective lens 55.

The deflected excitation light moves parallel and enters the AOTF 33. That is, the excitation light is incident on the AOTF 33 at the same incident angle as compared to before the deflection, and is incident on a different area of the AOTF 33.
The excitation light incident on the AOTF 33 enters the objective lens 55 and is collected on the subject 13.

  On the other hand, the stimulus light incident on the second scanning optical system 317 is deflected by the second scanning optical unit 29 and incident on the second relay lens system 331. The stimulation light incident on the second relay lens system 331 is incident on the AOTF 33 disposed at a position 33M conjugate with the focal position of the objective lens 55.

The deflected stimulation light is translated and incident on the AOTF 33. That is, the stimulation light is incident on the AOTF 33 at the same incident angle as compared with that before the deflection, and is incident on a different area of the AOTF 33.
The stimulation light incident on the AOTF 33 is diffracted, incident on the objective lens 55 together with the excitation light, and collected on the subject 13.

According to the above configuration, when the stimulation light is deflected, the incident angle of the stimulation light incident on the AOTF 33 does not change, and the incident position only moves in parallel. Therefore, compared with the case where the AOTF 33 is not disposed at the position 33M conjugate with the focal position of the objective lens 55, the incident position of the stimulation light is simply moved in parallel, so the optical path of the stimulation light is easily combined with the optical path of the excitation light. it can.
Since the allowable range of the incident angle of the light beam is narrow in the AOTF 33, the variation in the incident angle of the stimulation light is less than that in the case where the AOTF 33 is arranged at a position conjugate with the pupil of the objective lens 55. Can be easily combined with the optical path of the excitation light.

  Note that, as in the above-described embodiment, the stimulation light is only scanned in the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. Instead, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to the minute point.

[Fourth Embodiment]
Next, a fourth embodiment of the present invention will be described with reference to FIG.
The basic configuration of the laser scanning microscope of this embodiment is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIG. 8, and description of other components and the like will be omitted.
FIG. 8 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope in the present embodiment.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

As shown in FIG. 8, the laser scanning microscope 401 includes a first scanning optical system 15 into which excitation light is incident, an optical device 407, and an objective lens 55.
The optical device 407 deflects the stimulation light and introduces it into the optical path of the objective lens 55. The optical device 407 includes a second scanning optical system (stimulating light scanning optical system) 417 into which the first stimulation light and the second stimulation light are incident, and an AOD 34.

The second scanning optical system 417 deflects the incident first and second stimulation lights independently. The first and second stimulation lights are irradiated to a predetermined region of the subject 13 by being deflected by the second scanning optical system 417. The second scanning optical system 417 includes a second scanning optical unit 29 and a third scanning optical unit (stimulation light deflecting unit) 429.
Here, the first stimulation light and the second stimulation light are light having different wavelengths.

  The second scanning optical unit 29 deflects the first stimulation light so that the first stimulation light is incident on a predetermined region of the subject 13. On the other hand, the third scanning optical unit 429 makes the second stimulation light incident on a predetermined region of the subject 13 by deflecting the second stimulation light.

  Similarly to the second scanning optical unit 29, the third scanning optical unit 429 is provided with two third galvanometer mirrors (not shown), and the second stimulation light is supplied with two third galvanometers. It is deflected in one and other directions perpendicular to each other by the mirror.

  A control unit (signal generation unit) 443 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOD 34 is disposed in the AOD 34. The control unit 443 includes a signal generation unit 443A that generates a modulation signal that controls ultrasonic waves, and a high-frequency transmitter 443B that generates a high-frequency voltage based on the modulation signal. The high frequency transmitter 443B of the control unit 443 includes a first high frequency voltage for exciting the first ultrasonic wave having a frequency for diffracting the first stimulation light, and a second ultrasonic wave having a frequency for diffracting the second stimulation light. And a second high-frequency voltage that excites. The AOD 34 has a function of synthesizing both the first and second stimulation lights into the optical path of the first scanning optical system 315.

  In addition, the signal generation unit 443A of the control unit 443 generates a modulation signal that causes the high-frequency transmitter 443B to excite ultrasonic waves of the first and second frequencies.

Next, the operation of the laser scanning microscope 401 having the above configuration will be described.
Note that the description of the same parts as those of the first embodiment is omitted.
Further, the action of the first scanning optical system 15 on the excitation light is the same as that of the first embodiment, and the description thereof is omitted.

The first stimulation light incident on the second scanning optical system 417 of the optical device 407 is incident on the AOD 34 after being deflected by the second scanning optical unit 29 as shown in FIG. The incident angle of the first stimulation light to the AOD 34 is an incident angle related to the first-order diffraction direction of the AOD 34 at the wavelength of the first stimulation light.
On the other hand, the second stimulation light incident on the second scanning optical system 417 is deflected by the third scanning optical unit 429 and incident on the AOD 34. The incident angle of the second stimulation light to the AOTF 33 is an incident angle related to the first-order diffraction direction of the AOD 34 at the wavelength of the second stimulation light.

The stimulation light incident on the AOD 34 is diffracted, incident on the optical path of the objective lens 55 together with the excitation light, and collected on the subject 13.
Specifically, the control unit 443 inputs the first and second high-frequency voltages having frequencies corresponding to the first stimulation light and the second stimulation light to the transducer 37 of the AOD 34. The transducer 37 excites the crystal body 39 with first and second ultrasonic waves corresponding to the frequencies of the first and second high-frequency voltages. The crystal body 39 is formed with a periodic structure having a refractive index having a period corresponding to each stimulation light, and each stimulation light is diffracted by the AOD 34 and incident on the objective lens 55 together with the excitation light.

According to the above configuration, the AOD 34 can change the diffraction angle of the synthesized (diffracted) light according to the frequency of the ultrasonic wave (elastic wave) excited by the crystal body 39, so that one AOTF 33 Thus, the optical paths of the first stimulation light and the second stimulation light can be easily combined with the optical path of the excitation light.
By adjusting the intensity (amplitude) of the high-frequency voltage input to the AOTF 33, the specimen can be irradiated with the desired intensity of each of the first and second stimulation lights.

  By providing the second scanning optical unit 29 for deflecting the first stimulation light and the third scanning optical unit 429 for deflecting the second stimulation light, the first stimulation light and the second stimulation light are converted. It can be deflected separately and simultaneously focused and stimulated on different predetermined areas of the subject 13.

As in the above-described embodiment, the first stimulation light scans the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. In addition, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the first stimulation light may be irradiated to the minute point. Good.
Similarly, for the second stimulation light, the third scanning optical unit 429 not only scans the area to be stimulated in the subject 13 but also if the area to be stimulated is a minute point, You may make it irradiate 2 points | pieces of stimulation light to one minute point.

[Fifth Embodiment]
Next, a fifth embodiment of the present invention will be described with reference to FIG.
The basic configuration of the laser scanning microscope of this embodiment is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIG. 9, and description of other components and the like will be omitted.
FIG. 9 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope according to the present embodiment.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

As shown in FIG. 9, the laser scanning microscope 501 includes a first scanning optical system 15 into which excitation light is incident, an optical device 507, and an objective lens 55.
The optical device 507 deflects the stimulation light and introduces it into the optical path of the objective lens 55. The optical device 507 includes a second scanning optical system (stimulating light scanning optical system) 517 to which the first stimulation light and the second stimulation light are incident, and the optical paths of the deflected first and second stimulation lights. And an AOTF 33 that synthesizes the optical path of the polarized excitation light.

The second scanning optical system 517 deflects the incident first and second stimulation lights. The first and second stimulation lights are irradiated to a predetermined region of the subject 13 by being deflected by the second scanning optical system 517. The second scanning optical system 517 includes a second scanning optical unit 29 and a third scanning optical unit (stimulation light deflecting unit) 529.
Here, the first stimulation light and the second stimulation light are light having the same wavelength, and are one polarization and the other polarization of linearly polarized light orthogonal to each other. In the present embodiment, the first stimulus light is applied to s-polarized light and the second stimulus light is applied to p-polarized light.

The second scanning optical unit 29 deflects the first stimulation light so that the first stimulation light is incident on a predetermined region of the subject 13. On the other hand, the third scanning optical unit 529 makes the second stimulation light incident on a predetermined region of the subject 13 by deflecting the second stimulation light.
Similarly to the second scanning optical unit 29, the third scanning optical unit 529 is provided with two third galvanometer mirrors (not shown), and the second excitation light is supplied with two third galvanometers. It is deflected in one and other directions perpendicular to each other by the mirror.

  In the AOTF 33, a control unit 43 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOTF 33 is disposed. The high frequency transmitter 43B of the control unit 43 generates a high frequency voltage that excites an ultrasonic wave having a frequency that diffracts the first and second stimulation lights having the same frequency. Further, the signal generation unit 43A of the control unit 43 generates a modulation signal that causes the high frequency transmitter 43B to excite an ultrasonic wave having a frequency.

Next, the operation of the laser scanning microscope 501 having the above configuration will be described.
The description of the same parts as those in the first embodiment is omitted.
Further, the action of the first scanning optical system 15 on the excitation light is the same as that of the first embodiment, and thus the description thereof is omitted.

  As shown in FIG. 9, the first stimulation light incident on the second scanning optical system 517 of the optical device 507 is incident on the AOTF 33 after being deflected by the second scanning optical unit 29. The incident angle of the first stimulation light to the AOTF 33 is an incident angle related to the first-order diffraction direction of the AOTF 33 in the first stimulation light that is s-polarized light.

  On the other hand, the second stimulation light incident on the second scanning optical system 517 is deflected by the third scanning optical unit 529 and incident on the AOTF 33. The incident angle of the second stimulation light to the AOTF 33 is an incident angle related to the first-order diffraction direction of the AOTF 33 in the second stimulation light that is p-polarized light.

The stimulation light incident on the AOTF 33 is diffracted, incident on the objective lens 55 together with the excitation light, and collected on the subject 13.
Specifically, the control unit 43 inputs a high-frequency voltage having a frequency corresponding to the first and second stimulation lights, which are lights having the same frequency, to the transducer 37 of the AOTF 33. The transducer 37 excites an ultrasonic wave corresponding to the frequency of the high frequency voltage to the crystal body 39. The crystal body 39 is formed with a periodic structure having a refractive index having a period corresponding to each stimulation light, and each stimulation light is diffracted by the AOTF 33 and incident on the objective lens 55 together with the excitation light.

According to the above configuration, the first scanning optical system 15 and the second scanning optical system 517 are provided, and the AOTF 33 that combines both optical paths is provided. Therefore, a plurality of scanning operations can be performed without using a complicated configuration. The optical paths of the optical system can be easily synthesized.
The second scanning optical system 517 can deflect the incident first and second stimulation lights having the same wavelength, that is, can independently scan the respective stimulation lights having the same wavelength.

Since the AOTF 33 can cope with a change in the wavelength of the light beam to be synthesized (diffracted) by changing the frequency of the ultrasonic wave (elastic wave) excited by the crystal 39, the scanning optical system related to the same wavelength can be obtained by one AOTF 33. Easy optical path synthesis.
As a result, as compared with the optical system described in Patent Document 1 in which a plurality of polarization beam splitters corresponding to each wavelength needs to be arranged, a single AOTF 33 can synthesize a scanning optical system related to a plurality of wavelengths. The configuration of the optical device 507 can be simplified. Furthermore, in order to synthesize the optical paths of the plurality of scanning optical systems, the arrangement position of one AOTF 33 as compared with the optical system described in Patent Document 1 in which the arrangement angles of the plurality of polarizing beam splitters need to be adjusted. Since the optical paths of the plurality of scanning optical systems can be synthesized simply by adjusting the optical path etc., the optical apparatus 507 of this embodiment can facilitate the optical path synthesis.

  Since the second scanning optical system 517 includes the second scanning optical unit 29 that deflects the first stimulation light and the third scanning optical unit 529 that deflects the second stimulation light, the first scanning optical system 517 includes the first scanning optical unit 529. Stimulation light and second stimulation light can be operated separately. Therefore, a plurality of regions of the subject 13 can be stimulated simultaneously.

As in the above-described embodiment, the first stimulation light scans the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. In addition, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the first stimulation light may be irradiated to the minute point. Good.
Similarly, for the second stimulation light, the third scanning optical unit 529 not only scans the area to be stimulated in the subject 13 but also if the area to be stimulated is a minute point, You may make it irradiate 2 points | pieces of stimulation light to one minute point.

[Sixth Embodiment]
Next, a sixth embodiment of the present invention will be described with reference to FIG.
The basic configuration of the laser scanning microscope of this embodiment is the same as that of the first embodiment, but the configuration of the optical device is different from that of the first embodiment. Therefore, in this embodiment, only the periphery of the optical device will be described with reference to FIG. 10, and description of other components and the like will be omitted.
FIG. 10 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope in the present embodiment.
In addition, the same code | symbol is attached | subjected to the component same as 1st Embodiment, and the description is abbreviate | omitted.

As shown in FIG. 10, the laser scanning microscope 601 includes a first scanning optical system 15 on which excitation light is incident, an optical device 607, and an objective lens 55.
The optical device 607 deflects the stimulation light and combines the deflected optical path of the stimulation light and the deflected optical path of the excitation light. The optical device 607 includes a second scanning optical system (stimulating light scanning optical system) 617 on which stimulation light is incident, an AOTF 33, and an optical sensor (detection unit) 635.

  The second scanning optical system 617 deflects incident stimulation light. Stimulation light is deflected by the second scanning optical system 617 to irradiate a predetermined region of the subject 13. The second scanning optical system 617 includes a second scanning optical unit 29.

  The AOTF 33 is provided with a control unit (signal generation unit) 643 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOTF 33. The control unit 643 includes a signal generation unit 643A that generates a modulation signal for controlling ultrasonic waves, and a high-frequency transmitter 643B that generates a high-frequency voltage based on the modulation signal. In addition, the output signal of the optical sensor 635 is input to the signal generation unit 643A of the control unit 643.

The optical sensor 635 detects the intensity of the stimulation light transmitted through the AOTF 33 and outputs a detection signal based on the detected intensity of the stimulation light. The optical sensor 635 is disposed at a position where the stimulation light transmitted through the AOTF 33 is incident, and a detection signal output from the optical sensor 635 is input to the control unit 643 of the AOTF 33.
Note that a known sensor such as a photosensor can be used as the optical sensor 635 and is not particularly limited.

  A display unit 645 is connected to the optical sensor 635. The display unit 645 receives the detection signal from the optical sensor 635, and displays the intensity of the stimulation light introduced toward the objective lens 55 by converting the transmittance of the stimulation light to the optical sensor 635 side in the AOTF 33. To do. Currently, the operator can monitor the intensity of the stimulation light emitted toward the subject 13.

Next, the operation of the laser scanning microscope 601 having the above configuration will be described.
In addition, since the effect | action in parts other than the optical apparatus 607 of the laser scanning microscope 601 is the same as that of 1st Embodiment, the description is abbreviate | omitted.
In addition, the action of the optical device 607 on the excitation light is the same as that in the first embodiment, and a description thereof will be omitted.

  The stimulation light incident on the second scanning optical system 617 of the optical device 607 is incident on the AOTF 33 after being deflected by the second scanning optical unit 29 as shown in FIG. The incident angle of the stimulation light to the AOTF 33 is an incident angle related to the first-order diffraction direction of the AOTF 33 in the stimulation light. Most of the stimulation light incident on the AOTF 33 is diffracted, incident on the objective lens 55 together with the excitation light, and collected on the subject 13.

  On the other hand, part of the stimulation light incident on the AOTF 33 passes through the AOTF 33 without being diffracted and enters the optical sensor 635. The optical sensor 635 detects the intensity of the incident stimulation light and generates a detection signal corresponding to the intensity of the stimulation light. The generated detection signal is input to the power monitor 645 of the laser scanning microscope 601 and the control unit 643 of the AOTF 33.

  The control unit 643 generates a modulation signal for changing the voltage amplitude of the high-frequency voltage from the internal signal generation unit 643A based on the input detection signal. The control unit 643 generates a high frequency voltage whose voltage amplitude is controlled from the high frequency transmitter 643B based on the generated modulation signal. The generated high frequency voltage is input to the transducer 37 of the AOTF 33, and an ultrasonic wave having an amplitude corresponding to the voltage amplitude of the high frequency voltage is excited.

When the amplitude of the ultrasonic wave changes, the refractive index in the periodic structure of the refractive index formed in the crystal body 39 changes. Since the ratio of the stimulation light diffracted by the AOTF 33 and the stimulation light transmitted through the AOTF 33 changes with the change in the refractive index, the control unit 43 can control the intensity of the stimulation light incident on the subject 13.
Since the subsequent operation is the same as that of the first embodiment, the description thereof is omitted.

According to the above configuration, the control unit 643 generates a modulation signal based on the intensity of the stimulation light transmitted through the AOTF 33 detected by the optical sensor 635, and inputs the modulation signal to the AOTF 33, thereby stimulating the subject 13. Light intensity can be controlled freely.
For example, the optical device 607 of the present embodiment can control the intensity of the stimulation light incident on the objective lens 55 to be constant based on the intensity of the stimulation light detected by the optical sensor 635.

  Note that, as in the above-described embodiment, the stimulation light is only scanned in the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. Instead, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to the minute point.

[Seventh Embodiment]
Next, a seventh embodiment of the present invention will be described with reference to FIG.
The basic configuration of the laser scanning microscope of the present embodiment is the same as that of the second embodiment, but the configuration of the optical device is different from that of the second embodiment. Therefore, in the present embodiment, only the periphery of the optical device will be described with reference to FIG. 11, and description of other components and the like will be omitted.
FIG. 11 is a schematic diagram illustrating a configuration in the vicinity of the optical device of the laser scanning microscope according to the present embodiment.
Note that the same components as those of the second embodiment are denoted by the same reference numerals, and the description thereof is omitted.

As shown in FIG. 11, the laser scanning microscope 701 includes a first scanning optical system 215 that receives excitation light, an optical device 707, a third relay lens system 219, and an objective lens 55. .
The optical device 707 deflects incident stimulation light, and combines the deflected optical path of the stimulation light and the deflected optical path of the excitation light. The optical device 707 includes a second scanning optical system (stimulating light scanning optical system) 717 on which stimulation light is incident, and an AOTF 33.

  The second scanning optical system 717 deflects incident stimulation light. Stimulation light is deflected to the second scanning optical system 717 and is irradiated onto a predetermined region of the subject 13. The second scanning optical system 217 includes a second scanning optical unit (deflecting acousto-optic element) 729 and a second relay lens system 231.

The second scanning optical unit 729 makes the stimulation light incident on a predetermined area of the subject 13 by deflecting the stimulation light. The second scanning optical unit 29 is disposed between the second light source 5 and the second relay lens system 31 and at a position 29R conjugate with the pupil 11R of the objective lens 55.
Here, as the second scanning optical unit 729, an acousto-optic polarizer (hereinafter referred to as AOD) is used. The polarization direction of the stimulation light in the second scanning optical unit 729 is a direction orthogonal to the diffraction direction of the stimulation light in the AOTF 33 (in-plane direction perpendicular to the paper surface in FIG. 11).

  In the AOTF 33, a control unit (signal generation unit) 743 that controls ultrasonic waves (acoustic waves, elastic waves) excited by the crystal body 39 of the AOTF 33 is disposed. The control unit 743 includes a signal generation unit 743A that generates a modulation signal that controls ultrasonic waves, and a high-frequency transmitter 743B that generates a high-frequency voltage based on the modulation signal. Further, the signal generation unit 743A of the control unit 743 generates a modulation signal corresponding to the diffraction direction of the stimulation light.

Next, the operation of the laser scanning microscope 701 having the above configuration will be described.
In addition, since the effect | action in parts other than the optical apparatus 707 of the laser scanning microscope 701 is the same as that of 1st Embodiment, the description is abbreviate | omitted.
Further, the action of the optical device 707 on the excitation light is the same as that of the second embodiment, and thus the description thereof is omitted.

The stimulus light incident on the second scanning optical system 717 is deflected by the second scanning optical unit 729 and incident on the second relay lens system 231. The stimulation light incident on the second relay lens system 231 is incident on the AOTF 33.
The stimulus light incident on the AOTF 33 is diffracted in a predetermined direction, is incident on the objective lens 55 together with the excitation light, and is collected on the subject 13.
Since the subsequent operation is the same as that of the second embodiment, the description thereof is omitted.

According to the above configuration, the stimulation light is deflected in one direction intersecting with each other by the AOTF 33, and is deflected in the other direction intersecting with each other by the second scanning optical unit 729 which is an AOD. Therefore, the stimulation light can be scanned in one and other directions intersecting each other.
Since the deflected excitation light and stimulation light are incident on the AOTF 33, the deflected excitation light and stimulation light are incident on the objective lens 55 after being combined in an optical path, and are condensed on different predetermined regions of the subject 13. The Since the stimulation light is scanned in the one and other directions, the region where the stimulation light is collected in the subject 13 is scanned on the surface composed of the one and other directions.

  Note that, as in the above-described embodiment, the stimulation light is scanned not only in the area to be stimulated in the subject 13 by the second scanning optical unit 729 and the AOTF 33, but also the area to be stimulated is a minute point. For example, you may make it irradiate stimulation light to one minute point.

[Eighth Embodiment]
Next, an eighth embodiment of the present invention will be described with reference to FIG.
In this embodiment, the optical apparatus according to the present invention is applied to an optical microscope using normal epi-illumination.
The optical microscope 801 includes a microscope main body 805, an epi-illumination lamp house 803 that emits illumination light, an epi-illumination tube 813 that guides the illumination light toward the subject 13, and an optical device 807 that emits stimulation laser light. And an observation barrel 809 used for observation of observation light such as reflected light and fluorescence from the subject 13.

The microscope main body 805 includes an objective lens 55 arranged to face the subject 13 and a specimen stage 58 on which the subject 13 is placed.
The epi-illumination lamp house 803 is attached to the end of the epi-illumination tube 813. The epi-illumination lamp house 803 is provided with a mercury lamp (first light source) that emits illumination light.
The epi-illumination projection tube 813 includes a beam splitter 819 disposed on the microscope optical axis 801a, and is provided between the microscope main body 805 and the observation barrel 809.

  As the beam splitter 819, for example, a half mirror or a dichroic mirror can be used. The beam splitter 819 has a function of reflecting the illumination light from the epi-illumination lamp house 803. In addition, the function of transmitting the observation light from the subject 13 and the function of transmitting the stimulation light from the optical device 807 are also achieved.

The optical device 807 includes a second light source 5, a second scanning optical system 817, and an AOTF 33. The optical device 807 is attached as a single removable intermediate lens barrel unit between the incident light projection tube 813 and the observation lens barrel 809.
The second light source 5 emits stimulation laser light irradiated on the subject 13.
The second scanning optical system 817 deflects and scans stimulation laser light (hereinafter referred to as stimulation light), and includes a scanning optical unit 29 (specifically, a second galvanometer mirror 29A, 29B (see FIG. 1)) and a lens system (not shown).

  The AOTF 33 is arranged on the microscope optical axis 801a, and synthesizes the stimulation light of the optical device 807 with the optical path of the microscope. Since the AOTF 33 is controlled to diffract only the wavelength of the stimulation light, the observation light from the subject 13 is transmitted as it is.

The observation barrel 809 includes an imaging lens 56, an eyepiece optical system 847, and a switching optical system 845. On the optical path of the switching optical system 845, a camera 849 for capturing an image of the subject 13 is attached.
The imaging lens 56 forms an image so that an observer can visually recognize an image of observation light from the subject 13 or so that the camera 845 can capture an image of observation light.
The eyepiece optical system 847 enables the observer to visually recognize the observation light from the subject 13.

The switching optical system 845 selectively guides the observation light from the subject 13 to either the eyepiece optical system 847 or the camera 849.
The camera 849 images observation light from the subject 13.

Next, the operation of the microscope 801 having the above configuration will be described.
Illumination light emitted from the first light source 803 enters a beam splitter 819 as shown in FIG. The illumination light incident on the beam splitter 819 is reflected toward the objective lens 55.

On the other hand, the stimulation light emitted from the second light source 5 is incident on the second scanning optical unit 29 and deflected in a predetermined direction by the second scanning optical unit 29.
The deflected stimulation light is incident on the AOTF 33 from the first-order diffraction direction of the AOTF 33. The stimulus light incident on the AOTF 33 is diffracted and emitted toward the beam splitter 819. The stimulus light passes through the beam splitter 819 and enters the objective lens 55 through the same optical path as the illumination light described above.

The illumination light and the stimulation light incident on the objective lens 55 are collected on the subject 13, respectively.
The illumination light is applied to the entire observation region of the subject 13, while the stimulation light is deflected and scanned by the second scanning optical unit 29. For example, the stimulation light is scanned so as to irradiate only a predetermined area in the subject 13.

Illumination light reflected from the subject 13 and fluorescence excited by the illumination light (hereinafter referred to as fluorescence or the like) propagates in the opposite direction of the optical path through which the stimulation light propagates and enters the observation tube 809. To do.
Specifically, fluorescence or the like first enters the objective lens 55 and then enters the beam splitter 819 from the objective lens 55. The fluorescence or the like passes through the beam splitter 819 and enters the AOTF 33. The fluorescence or the like incident on the AOTF 33 has a wavelength different from that of the stimulus light, and thus passes through the AOTF 33 and enters the observation barrel 809. The reflected stimulation light is diffracted by the AOTF 33 and therefore does not enter the observation barrel 809.

  The fluorescence or the like that has passed through the AOTF 33 and entered the observation barrel 809 enters the switching optical system 845. Fluorescence and the like are incident on the eyepiece optical system 847 or the camera 849 by the switching optical system 845. The fluorescence or the like incident on the eyepiece optical system 847 is observed by an observer, and the fluorescence or the like incident on the camera 849 is captured by the camera 849. Note that the switching optical system 845 selects an incident destination of fluorescence or the like by an operation of an observer or the like.

According to the above configuration, the optical device 807 according to this embodiment can be easily attached to and detached from the optical microscope 801 that irradiates the subject 13 with illumination light. Therefore, the optical device 807 is simply attached to the optical microscope 801. Thus, an observation method for observing the reaction of the subject 13 by irradiating the desired position or region of the subject 13 with the stimulation light can be easily executed.
Since the optical device 807 is provided with the second scanning optical system 817 and the AOTF 33, the stimulation light can be scanned and condensed at a predetermined position or region of the subject 13.

  Note that, as in the above-described embodiment, the stimulation light is only scanned in the region to be stimulated in the subject 13 by the second galvanometer mirrors 29A and 29B (see FIG. 1) of the second scanning optical unit 29. Instead, if the region to be stimulated is a minute point, the second galvanometer mirrors 29A and 29B may be fixed at a desired position and the stimulation light may be irradiated to the minute point.

FIG. 13 is a modification of the present embodiment.
As shown in FIG. 13, the optical device 807 ′ may be attached to the incident light projection tube 813 ′. In this case, the AOTF 33 is arranged in the optical path of the epi-illumination in the epi-illumination projection tube 813 ′. However, since the AOTF 33 is controlled so as to diffract only the wavelength of the stimulation light, the epi-illumination lamp house 803 The incident illumination light passes through the AOTF 33 as it is.

  Further, as the beam splitter 819, a beam that reflects the stimulation light and the epi-illumination light and transmits the observation light is used, and examples thereof include a half mirror and a dichroic mirror.

The technical scope of the present invention is not limited to the above embodiment, and various modifications can be made without departing from the spirit of the present invention.
In the above-described embodiment, the optical device according to the invention of the present application has been described with reference to a microscope, but the present invention can also be applied to various other optical devices.

  Note that the present invention may be applied to the above-described embodiments, and the fourth embodiment, the fifth embodiment, and the sixth embodiment are the second embodiment or the third embodiment. It may be implemented in combination with the form, and is not particularly limited.

  The first scanning optical system (observation optical system) 15, 215, 315 includes a first scanning optical unit (scanning optical system) 21 using a galvanometer mirror as in the above-described embodiments. A focus detection optical system may be used, or a disk scanning optical system that scans light by rotating a disk having a large number of minute openings such as pinholes and slits, and is not particularly limited.

It is the schematic explaining the structure of the laser scanning microscope which concerns on the 1st Embodiment of this invention. It is a schematic diagram explaining the structure of the optical apparatus vicinity of FIG. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 1st modification of 1st Embodiment in this invention. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 2nd Embodiment of this invention. It is a schematic diagram which shows the state of the light ray in the optical apparatus of FIG. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 3rd Embodiment of this invention. It is a schematic diagram which shows the state of the light ray in the optical apparatus of FIG. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 4th Embodiment of this invention. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 5th Embodiment of this invention. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 6th Embodiment of this invention. It is a schematic diagram explaining the structure of the optical apparatus vicinity of the laser scanning microscope in the 7th Embodiment of this invention. It is a schematic diagram explaining the structure of the microscope which concerns on the 8th Embodiment of this invention. It is a schematic diagram explaining the structure of the modification of 8th Embodiment.

Explanation of symbols

7, 107, 207, 307, 407, 507, 607, 707, 807 Optical device 13 Subject 15, 215, 315 First scanning optical system (observation optical system)
17, 117, 217, 317, 417, 617, 717, 817 Second scanning optical system (stimulating light scanning optical system)
21 First scanning optical unit (scanning optical system)
29 Second scanning optical unit (stimulating light deflection unit)
33 Acoustooptic modulation filter, AOTF (synthetic acoustooptic device)
43, 143, 443, 643, 743 Control unit (signal generation unit)
43A, 143A, 443A, 643A, 743A Signal generator 55 Objective lens (objective optical system)
429, 529 Third scanning optical unit (stimulating light deflection unit)
635 Optical sensor (detector)
729 Second scanning optical unit (deflection acousto-optic device)

Claims (14)

A scanning optical system for stimulation light that deflects stimulation light having a wavelength of at least one or more, which is stimulation light that changes a subject;
A synthetic acousto-optic device that synthesizes the optical path of the deflected stimulus light with the optical path of the observation optical system for obtaining the image of the subject;
An optical device provided with   The optical apparatus according to claim 1, wherein the stimulation light scanning optical system and the synthetic acousto-optic element can be inserted into and removed from an optical path of the observation optical system.   The optical apparatus according to claim 1, wherein the stimulation light scanning optical system is provided with a stimulation light polarization unit that deflects the stimulation light. The synthetic acoustooptic element deflects the stimulation light in one direction intersecting each other;
The stimulation light scanning optical system is provided with a polarization acousto-optic element that deflects the stimulation light in other directions intersecting each other.
4. The optical device according to claim 1, wherein the synthetic acoustooptic element receives the stimulus light deflected by the deflecting acoustooptic element and the other light beam. 5.   The optical apparatus according to claim 1, wherein the synthetic acoustooptic element diffracts the stimulation light and transmits light in an optical path of the observation optical system. An objective optical system for condensing the stimulation light on the subject;
The optical apparatus according to claim 1, wherein the synthetic acoustooptic element is disposed at a position conjugate with a pupil of the objective optical system. An objective optical system for condensing the stimulation light on the subject;
The optical apparatus according to claim 1, wherein the synthetic acoustooptic element is disposed at a position conjugate with an image of the objective optical system. A signal generation unit for generating a modulation signal for controlling an ultrasonic wave excited in the synthetic acoustooptic device;
The optical apparatus according to claim 1, wherein the signal generation unit controls the intensity of the deflected stimulation light irradiated on the subject by controlling the ultrasonic wave. A detection unit for detecting the intensity of the deflected stimulation light that passes through the synthetic acoustooptic device without being irradiated on the subject;
The optical apparatus according to claim 8, wherein the signal generation unit generates the modulation signal based on the intensity of the stimulation light detected by the detection unit.   The optical apparatus according to claim 1, wherein the synthetic acoustooptic element is an acoustooptic modulation filter.   The optical apparatus according to claim 1, wherein the synthetic acoustooptic element is an acoustooptic modulator.   The optical apparatus according to claim 1, wherein the synthetic acoustooptic element is an acoustooptic deflector. In any one of claims 1 to 12,
An optical apparatus in which the observation optical system includes a scanning optical system that scans light that irradiates the subject. In any one of claims 1 to 12,
An optical apparatus in which the observation optical system is a non-scanning microscope optical system.

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