JP2008175884A - Laser microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To detect the intensity of a laser beam emitted from the front end of an objective lens with simplicity and satisfactory accuracy in spite of a change in the wavelength and intensity of the laser beam and to irradiate a sample with a laser beam of the accurately set intensity. <P>SOLUTION: There is provided the laser microscope, which includes a laser light source 13 emitting a high-output laser beam L<SB>4</SB>and an objective lens 20 irradiating the sample A with the laser beam L<SB>5</SB>emitted from the laser light source 13, wherein a beam splitter 33 arranged between the laser light source 13 and the objective lens 20 and branching a portion of the laser beam L<SB>5</SB>from the laser light source 13, a light quantity detecting means 34 detecting the light quantity of the laser beam L3 branched from the beam splitter 33, and a diaphragm device 35 arranged between the light quantity detecting means 34 and the beam splitter 33 and limiting a luminous flux diameter are provided. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a laser microscope.

Conventionally, a part of the laser light from the laser light source that combines and emits laser light of different wavelengths is separated by a beam splitter, the wavelength is selected by an exchangeable filter, and the laser light of the selected wavelength is detected There is known a laser microscope that receives light by a detector, detects the intensity of laser light of that wavelength, and adjusts the intensity of the laser light based on the detection signal (see, for example, Patent Document 1).
In this laser microscope, the intensity of the laser light emitted from the objective lens can be known by considering the transmittance and reflectance for each wavelength from the position of the photodetector to the objective lens.

JP-A-11-231222

However, PCF (Photonic Crystal) capable of broadband single-mode transmission in the laser light transmission path.
When using (Fiber), the numerical aperture (NA) of the laser light emitted from the PCF is proportional to the wavelength. For this reason, when the laser light emitted from the PCF is made to be substantially parallel light using a collimator lens, the beam diameter of the long wavelength laser light becomes larger than the beam diameter of the short wavelength laser light. is there.

  In addition, when a high-power laser light source that emits near-infrared ultrashort pulse laser light for multiphoton excitation is used, the beam diameter of the laser light also changes due to the thermal lens effect of the optical system that guides . The greater the power of the laser light source and the closer the wavelength is to the near-infrared region, the more easily the thermal lens effect is caused, and the beam diameter changes greatly.

In this case, if the beam diameter of the laser light changes due to a change in the wavelength of the laser light or a thermal lens effect, the beam diameter may become larger than the pupil diameter of the objective lens.
When the beam diameter is larger than the pupil diameter of the objective lens, the outer part of the laser beam is displaced by the pupil of the objective lens, so the intensity of the laser beam actually emitted from the tip of the objective lens is incident on the photodetector. Therefore, there is a problem that the intensity of the laser beam is reduced and the accurate intensity cannot be known.

  The present invention has been made in view of the above-described circumstances, and can easily and accurately detect the intensity of laser light emitted from the tip of an objective lens even if the wavelength or intensity of the laser light changes. An object of the present invention is to provide a laser microscope capable of irradiating a specimen with laser light having a precisely set intensity.

In order to achieve the above object, the present invention provides the following means.
The present invention comprises a laser light source that emits high-power laser light, and an objective lens that irradiates a sample with laser light emitted from the laser light source, and is disposed between the laser light source and the objective lens, A beam splitter for branching a part of the laser light from the laser light source, a light amount detecting means for detecting the light amount of the laser light branched by the beam splitter, and disposed between the light amount detecting means and the beam splitter, Provided is a laser microscope including a diaphragm device for limiting a beam diameter.

  According to the present invention, laser light emitted from a laser light source is irradiated onto a specimen by an objective lens, whereby a fluorescent substance contained in the specimen is excited and fluorescence is generated. By detecting this fluorescence, a fluorescence image of the specimen can be observed. A part of the laser light emitted from the laser light source is branched by the beam splitter and detected by the light amount detecting means. Thereby, the intensity of the laser light irradiated from the objective lens to the specimen can be grasped.

  In this case, when the laser light emitted from the laser light source has a high output, the beam diameter of the laser light changes due to the thermal lens effect of the optical system that guides the light. When the beam diameter of the laser beam is enlarged, the beam diameter of the laser beam emitted from the objective lens is such that the aperture element disposed on the optical path including the objective lens from the beam splitter, for example, the pupil of the objective lens, etc. May be limited by.

  According to the present invention, when the light beam diameter of the laser beam branched by the beam splitter is larger than the predetermined diameter dimension, the light beam diameter is limited by the diaphragm device arranged between the light amount detecting means and the beam splitter. After that, it is detected by the light quantity detection means. Therefore, by making the aperture device coincide with the aperture element, even if the thermal lens effect or the like occurs and the beam diameter fluctuates, the light amount of the laser light emitted from the objective lens can be accurately detected by the light amount detection means. Can be detected.

  The present invention also includes a laser light source capable of emitting laser beams of different wavelengths, and an objective lens that irradiates the specimen with the laser light emitted from the laser light source, and is disposed between the laser light source and the objective lens. A beam splitter for branching a part of the laser light from the laser light source, a light quantity detection means for detecting the light quantity of the laser light branched by the beam splitter, and between the light quantity detection means and the beam splitter Provided is a laser microscope provided with a diaphragm device that is arranged and restricts a beam diameter.

  According to the present invention, laser light emitted from a laser light source is irradiated onto a specimen by an objective lens, whereby a fluorescent substance contained in the specimen is excited and fluorescence is generated. By emitting laser light with a plurality of wavelengths from the laser light source, it becomes possible to detect fluorescence in a plurality of wavelength bands, and more detailed fluorescence observation of the specimen can be performed.

In this case, when the laser light emitted from the laser light source covers a wide wavelength band, the exit numerical aperture of the optical system that guides the light, for example, a photonic crystal fiber (PCF), is proportional to the wavelength. The beam diameter changes.
According to the present invention, when the light beam diameter of the laser beam branched by the beam splitter is larger than the predetermined diameter dimension, the light beam diameter is limited by the diaphragm device arranged between the light amount detecting means and the beam splitter. After that, it is detected by the light quantity detection means. Therefore, by making the diaphragm device coincide with the diaphragm element on the optical path such as the pupil of the objective lens, the light quantity of the laser light emitted from the objective lens can be accurately detected by the light quantity detection means even if the beam diameter varies. Can be detected.

In the above invention, it is preferable that the beam splitter is disposed between the objective lens and an imaging lens that forms an image of light collected by the objective lens.
By doing so, the laser light for light quantity detection can be branched at a position close to the objective lens where the beam diameter of the laser light from the laser light source is limited, and the light quantity of the laser light emitted from the objective lens can be more accurately It becomes possible to detect.

Moreover, in the said invention, it is preferable that the said diaphragm | throttle device is arrange | positioned in the optically equivalent position with the pupil position of the said objective lens.
In this way, it is possible to limit the light beam diameter of the laser light detected under the same conditions as the pupil of the objective lens where the light beam diameter of the laser light is limited, and the laser light irradiated from the objective lens to the specimen Can be detected with higher accuracy.

Moreover, in the said invention, the said beam splitter is good also as being provided in the optical path between the said laser light source and the said objective lens so that attachment or detachment is possible.
By doing so, a beam splitter is arranged on the optical path when the light amount is detected and the laser light is branched, and the beam splitter is removed from the optical path when observing, thereby eliminating the waste of the laser light irradiated on the specimen. Can do.

In the above invention, the diaphragm device may have an aperture shape substantially the same as the aperture shape of the pupil position of the objective lens.
In this way, the beam diameter of the laser beam for detection can be limited under the same conditions as the limitation on the pupil position of the objective lens, and the amount of laser light emitted from the objective lens to the sample can be more accurately controlled. Can be detected.

In the above invention, the aperture diameter of the diaphragm device is a dimension obtained by multiplying the aperture diameter of the pupil position of the objective lens by the magnification of the optical system arranged between the objective lens and the beam splitter. It is good.
By doing in this way, even if the expansion or reduction optical system is provided between the objective lens and the beam splitter, the light quantity of the laser light irradiated on the specimen from the objective lens can be detected with high accuracy.

Moreover, in the said invention, it is good also as the said aperture_diaphragm | restriction apparatus being provided so that the opening diameter can be changed.
In this way, when the objective lens is exchanged, the aperture diameter of the aperture device is changed, and the laser beam irradiated to the specimen from the objective lens is similarly applied to the objective lens having a different aperture diameter. Can be detected with high accuracy.

Moreover, in the said invention, the said objective lens is provided interchangeably and it is good also as providing the adjustment apparatus which adjusts the aperture diameter of the said aperture device according to the said objective lens arrange | positioned on an optical axis. .
In this way, when the objective lens is exchanged, the aperture diameter of the diaphragm device is adjusted by the operation of the adjusting device. Therefore, the objective lens is changed from the objective lens to the specimen regardless of the type of the objective lens arranged on the optical axis. It is possible to accurately detect the amount of laser light to be irradiated.

  According to the present invention, even when the wavelength or intensity of the laser beam changes, the intensity of the laser beam emitted from the tip of the objective lens can be detected easily and accurately, and the laser having a precisely set intensity. There is an effect that the specimen can be irradiated with light.

A laser microscope 1 according to an embodiment of the present invention will be described below with reference to FIG.
As shown in FIG. 1, the laser microscope 1 according to this embodiment includes a laser light source 2, a microscope main body 3, and a single mode fiber 4 such as a photonic crystal fiber that connects the laser light source 2 and the microscope main body 3. And a control unit 5.

The laser light source 2 includes a first light source unit 6 and a second light source unit 7.
The first light source unit 6 includes an argon laser light source 8 capable of simultaneously oscillating laser light L ₁ having a plurality of wavelengths λ ₁ (for example, 458 nm, 488 nm, 514.5 nm), and a laser having a wavelength λ ₂ (for example, 405 nm). The laser diode 9 that oscillates the light L ₂ , the dichroic mirror 10 that combines the laser light L ₁ and L ₂ emitted from the argon laser light source 8 and the laser diode 9, and the acoustooptic device 11 are provided. In the figure, reference numeral 12 denotes a mirror.
Acousto-optic device 11, based on a signal from the control unit 5, the wavelength selection of the laser beam L ₃ is emitted towards the microscope main body 3, and performs switching of the intensity modulation and off.

The second light source unit 7 includes a titanium sapphire laser light source (high-power laser light source) 13 that oscillates the optimized femtosecond laser light L ₄ over a wide range of 690 to 1040 nm, and the control unit 5. based on the signal and an acousto-optic device 14 for the switching of the intensity modulation and on-off of the laser beam L ₅ emitted toward the microscope body 3.

The laser light L ₃ from the first light source unit 6 is guided to the microscope main body 3 through the single mode fiber 4, and the laser light L ₅ from the second light source unit 7 is directly applied to the microscope main body 3 through the spatial light path. It has come to be guided.

The microscope body 3 includes a mirror 15 and a dichroic mirror 16 that allow the laser beams L ₃ and L ₅ from the first light source unit 6 and the second light source unit 7 to enter the same optical path, and a laser from the laser light source 2. a scanner 17 for scanning the light _L 3, _{L 5} two-dimensionally, a pupil projection lens 18 for condensing the laser beam _L 3, _{L 5} scanned by the scanner 17, is focused by the pupil projection lens 18 an imaging lens 19 for converting the laser beam L ₃ forming the intermediate _image, L ₅ a substantially parallel light, the laser light L _3, L _5, which is converted into substantially parallel light by said imaging lens 19 condenses The objective lens 20 that irradiates the specimen A while condensing the fluorescence F returning from the specimen A, and the fluorescence F that is collected by the objective lens 20 and returns via the imaging lens 19, the pupil projection lens 18, and the scanner 17. The A dichroic mirror 21 that branches from the optical path of the laser light L _3, L _5, and a light detection unit 22 for detecting the fluorescence F branched by the dichroic mirror 21.

The light detection unit 22 includes a condensing lens 23 that condenses the fluorescence F branched from the laser beams L ₃ and L ₅ by the dichroic mirror 21, and a confocal pinhole 24 disposed at the focal position of the condensing lens 23. A collimating lens 25 that makes the fluorescent light F that has passed through the confocal pinhole 24 substantially parallel light, a dichroic mirror 26 that branches the fluorescent light F that has become substantially parallel light for each wavelength, and the dichroic mirror 26. And a plurality of photodetectors 27 and 28 such as photomultiplier tubes for detecting the fluorescence F for each wavelength.

In the figure, reference numerals 29 and 30 are mirrors, and reference numeral 31 is a barrier filter. The confocal pinhole 24 is disposed at a position optically conjugate with the focal plane of the objective lens 20.
In addition, a dichroic mirror 36 is provided between the pupil projection lens 18 and the imaging lens 19. Of the fluorescence F condensed by the imaging lens 19, the laser beam L ₅ from the titanium sapphire laser light source 13 is used. excited fluorescence F ₁ is adapted to branch. The branched fluorescence F ₁ is detected by the photodetector 37.

  As the objective lens 20, one objective lens 20 is selected from a plurality of objective lenses 20 arranged on the turret 32 according to the magnification and the like. Information on the selected objective lens 20 is sent to the control unit 5.

In the present embodiment, a beam splitter 33 is arranged between the dichroic mirrors 16 and 21 so as to branch a part of the laser beams L ₃ and L ₅ from the laser light source 2. Further, a light amount detector 34 such as a photodiode for detecting the laser beams L ₃ and L ₅ branched by the beam splitter 33 is provided. Further, in the present embodiment, a variable diaphragm (a diaphragm device) 35 is disposed between the beam splitter 33 and the light amount detector 34.

The control unit 5 outputs to the acoustooptic elements 11 and 14 of the light source units 6 and 7 wavelength selection and on / off switching commands of the laser beams L ₁ , L ₂ and L ₄ from the light sources 8, 9 and 13. It is supposed to be. The control unit 5 receives the light quantity information of the laser lights L ₃ and L ₅ detected by the light quantity detector 34, controls the acoustooptic elements 11 and 14, and emits the laser beams from the light source units 6 and 7. Light intensity modulation of the light L ₃ and L ₅ is performed.

  The control unit 5 adjusts the aperture diameter of the diaphragm device 35 based on information on the objective lens 20 sent from the turret 32 on which the objective lens 20 is mounted. Preferably, the aperture diameter of the diaphragm device 35 is set to a value obtained by multiplying the pupil diameter of the objective lens 20 by the magnifications of the pupil projection lens 18 and the imaging lens 19. The aperture shape of the diaphragm device 35 is preferably the same as the shape of the diaphragm arranged at the pupil position of the objective lens 20.

The operation of the laser microscope 1 according to this embodiment configured as described above will be described below.
In order to observe the specimen A using the laser microscope 1 according to the present embodiment, first, the laser light L ₁ (or laser light L ₂ ) is emitted from the first light source unit 6. The emitted laser beams L ₁ and L ₂ are incident on the microscope main body 3 through the single mode fiber 4 as the laser beam L ₃ adjusted by the acoustooptic device 11. In the microscope main body 3, the laser light L ₃ is reflected by the mirror 15 and the dichroic mirror 16, and then scanned two-dimensionally by the scanner 17, via the pupil projection lens 18, the imaging lens 19 and the objective lens 20. The specimen A on the stage 38 is irradiated.

Fluorescence F generated in the specimen A by being irradiated with the laser beam L ₃ are condensed by the objective lens 20, back through the imaging lens 19, the dichroic mirror 36, the pupil projection lens 18 and the scanner 17, It is branched from the optical path of the laser beam L ₃ by the dichroic mirror 21. The branched fluorescence F is condensed by the condensing lens 23 and only the light that has passed through the confocal pinhole 24 is detected by the photodetector 27 (through the mirror 29, the collimating lens 25, and the dichroic mirror 26 (and the mirror 30)). Alternatively, it is detected by a photodetector 28). The laser light L _{3 that} has not been separated by the dichroic mirror 21 and has transmitted with the fluorescence F is blocked by the barrier filter 31.

  Since the confocal pinhole 24 is provided, only the fluorescence F emitted from the focal plane of the objective lens 20 can pass through the confocal pinhole 24 and is detected by the photodetector 27. Therefore, based on the intensity of the fluorescence F detected by the photodetector 27 and the scanning position information of the scanner 17 at the time when the fluorescence F is detected, it is two-dimensionally set to a predetermined depth in the specimen A along the focal plane. It is possible to acquire a clear fluorescent image on a plane that spreads out.

Further, when performing multiphoton excitation observation of the specimen A, the laser light L ₄ is emitted from the titanium sapphire laser light source 13 of the second light source unit 7. The emitted laser beam L ₄ is incident on the microscope main body 3 as the laser beam L ₅ adjusted by the acoustooptic device 14. In the microscope main body 3, the laser beam L ₅ passes through the dichroic mirror 16, and then is scanned two-dimensionally by the scanner 17, and then on the stage 38 via the pupil projection lens 18, the imaging lens 19 and the objective lens 20. The specimen A is irradiated.

The fluorescence F ₁ generated in the specimen A by being irradiated with the laser light L ₅ is collected by the objective lens 20, then returns through the imaging lens 19, and is removed from the optical path of the laser light L ₅ by the dichroic mirror 36. Branch off. The branched fluorescence F ₁ is detected by the photodetector 37.

In the multi-photon excitation observation, since the fluorescence F ₁ is generated only at the condensing position of the laser light L ₅ , the intensity of the fluorescence F ₁ detected by the photodetector 37 and the scanner at the time when the fluorescence F ₁ is detected. Based on the 17 scan position information, it is possible to acquire a clear multiphoton fluorescence image in a plane that extends two-dimensionally to a predetermined depth in the specimen A along the focal plane.

In this case, a part of the laser beams L ₃ and L ₅ from the light source units 6 and 7 are branched by the beam splitter 33 disposed between the dichroic mirrors 16 and 21 and detected by the light amount detector 34. Therefore, if the transmittance of the beam splitter 33 is known, the intensities of the laser beams L ₃ and L ₅ irradiated from the objective lens 20 to the sample A are determined based on the detected light amounts of the laser beams L ₃ and L ₅ . It becomes possible to recognize correctly. Furthermore, in the present embodiment, the light amount is detected when the beam diameters of the laser beams L ₃ and L ₅ become greater than or equal to a predetermined value due to the operation of the diaphragm device 35 disposed between the beam splitter 33 and the light amount detector 34. The beam diameters of the laser beams L ₃ and L ₅ incident on the device 34 are limited.

Since the aperture diameter of the aperture device 35 is set to a value obtained by multiplying the pupil diameter of the objective lens 20 by the magnification of the pupil projection lens 18 and the imaging lens 19, the laser beams L ₃ and L incident on the aperture device 35 are set. the proportion of the laser beam L _3, L ₅ incident on the light quantity detector 34 for _5, equal to the ratio of the laser beam L _3, L ₅ incident on the specimen with respect to the laser beam L ₃ incident on the objective lens 20 be able to. That is, when the light beam diameters of the laser beams L ₃ and L ₅ incident on the objective lens 20 are larger than the pupil diameter of the objective lens 20, the outer peripheral portion of the light beam is displaced by the diaphragm arranged at the pupil position, and actually In addition, the light amounts of the laser beams L ₃ and L ₅ irradiated to the specimen A are smaller than the incident light amount to the objective lens 20. In the present embodiment, similarly to the case where the laser beams L ₃ and L ₅ irradiated to the specimen A are limited by the diaphragm arranged at the pupil position of the objective lens 20, the laser beams incident on the light amount detector 34. By restricting the light amounts of L ₃ and L ₅ by the diaphragm device 35, there is an advantage that the light amounts of the laser beams L ₃ and L ₅ that are actually irradiated on the specimen A can be accurately grasped.

For example, even when the turret 32 is fixed and the objective lens 20 is not changed, the laser light L emitted from the single mode fiber 4 in proportion to the wavelength of the laser light L ₃ emitted from the light source unit 6. _Since the numerical aperture of ₃ changes, the beam diameter of the laser beam L ₃ incident on the objective lens 20 changes. Further, the beam diameter of the laser beam L ₅ emitted from the high-power titanium sapphire laser light source 13 fluctuates due to the thermal lens effect of the optical system that passes through to the objective lens 20.

In this embodiment, even in such a case, the light beam diameters of the laser beams L ₃ and L ₅ incident on the light amount detector 34 at the same rate as the light beam diameter is limited at the pupil position of the objective lens 20. Therefore, there is an advantage that the laser beams L ₃ and L ₅ irradiated on the specimen A can be accurately grasped.
The light amounts of the laser beams L ₃ and L ₅ detected by the light amount detector 34 are sent to the control unit 5, and the acoustooptic device is feedback-controlled based on the light amount information, so that a laser having a desired light amount is obtained. The specimen L can be irradiated with the light L ₃ and L ₅ accurately and stably.

Moreover, since the wavelength of the titanium sapphire laser light source 13 is variable in the wavelength band 690 to 1040 nm, the excitation spectrum of the sample A is obtained by changing the wavelength of the laser light L ₄ and acquiring the fluorescence luminance value from the sample A. Can be obtained. At the time of obtaining the excitation spectrum, changing wavelength, there must be the amount of the laser beam L _4, which is emitted from the objective lens 20 is constant.

According to the present embodiment, even if the light beam diameter of the laser beam L ₄ emitted from the objective lens 20 varies due to the change in wavelength and the beam diameter of the laser beam L ₄ changes, the laser beam is emitted from the objective lens 20. amount can accurately measure that is, can the light amount of the laser beam L ₄ constant by controlling the acousto-optic element 14 with the measurement result. As a result, the excitation spectrum of the specimen A can be obtained accurately.

In the present embodiment, since the control unit 5 adjusts the aperture diameter of the diaphragm device 35 based on the information of the objective lens 20 sent from the turret 32, the objective lens 20 having a different pupil diameter is used. Also, the laser beam L irradiated to the specimen A is limited by limiting the beam diameters of the laser beams L ₃ and L ₅ detected by the light amount detector 34 at the same rate as the beam diameter limited by the objective lens 20. ₃ and L ₅ can always be set with high accuracy.

In the present embodiment, a part of the laser beams L ₃ and L ₅ is split between the dichroic mirrors 16 and 21 by the beam splitter 3 for light quantity detection. Instead, as shown in FIG. Alternatively, the light quantity may be measured using the laser beams L ₃ and L ₅ branched immediately before the objective lens 20. In this way, the diaphragm device 35 can be disposed at a position optically equivalent to the pupil position of the objective lens 20, and the sample from the objective lens 20 can be accurately obtained without being affected by other optical systems. The light quantity of the laser beams L ₃ and L ₅ irradiated to A can be measured.

  In the present embodiment, a variable diaphragm is used as the diaphragm device 35. Instead of this, an opening prepared for each objective lens 20 and a turret type or slide type switching mechanism (for switching the opening) ( (Not shown) may be adopted. An objective lens may be adopted as the diaphragm device 35.

Moreover, it is preferable that the beam splitter 33 is provided so as to be detachable with respect to the optical paths of the laser beams L ₃ and L ₅ . By doing so, the beam splitter 33 is inserted into the optical path when measuring the amount of light, and the beam splitter 33 is removed from the optical path when observing the specimen A, so that all the laser beams L ₃ . The L ₅ can be used efficiently observed. In this case, instead of the beam splitter 33, a mirror may be employed.

  Further, although the diaphragm device 35 is disposed between the beam splitter 33 and the light amount detector 34, it may be disposed in front of the beam splitter 33 instead. By doing in this way, compared with the case where it arrange | positions in the front | former stage of the light quantity detector 34, the problem that the flare which generate | occur | produced in the aperture device 35 injects into a light quantity detector is prevented, and the precision of light quantity detection is improved. There is an advantage that you can.

Further, in the present embodiment, the diaphragm device 35 including a variable diaphragm is employed in order to cope with the objective lens 20 having a different pupil diameter. Instead, a fixed diaphragm having a predetermined aperture diameter and the fixed diaphragm are used. You may decide to employ | adopt the zoom mechanism (not shown) which changes the light beam diameter of the laser beams L3 and L5 which enter into a stop.
Further, instead of an acoustooptic element such as AOM or AOTF that modulates the laser beams L ₁ , L ₂ , and L ₄ , an electro-optic element or a magneto-optic element may be employed.

  In this embodiment, the case where the beam diameter is limited by the aperture element arranged at the pupil position of the objective lens 20 has been described. Instead, the beam diameter is limited by a metal frame or the like other than the pupil position. In this case, the diaphragm device 35 may be arranged at a position equivalent to the metal frame or the like.

1 is a block diagram illustrating an overall configuration of a laser microscope according to an embodiment of the present invention. It is a figure which shows the modification of the laser microscope of FIG.

Explanation of symbols

A sample _{L 1, L 2, L 3} , L 4, L 5 laser beam first laser microscope 5 control unit (adjusting device)
8 Argon laser light source (laser light source)
9 Laser diode (laser light source)
13 Titanium sapphire laser light source (laser light source)
DESCRIPTION OF SYMBOLS 19 Imaging lens 20 Objective lens 33 Beam splitter 34 Light quantity detector (light quantity detection means)
35 Aperture device

Claims (9)

A laser light source that emits high-power laser light, and an objective lens that irradiates the sample with laser light emitted from the laser light source,
A beam splitter that is arranged between the laser light source and the objective lens and branches a part of the laser light from the laser light source; a light amount detecting means for detecting the light amount of the laser light branched by the beam splitter; A laser microscope comprising: a diaphragm device disposed between the light amount detection means and the beam splitter, for limiting a beam diameter. A laser light source capable of emitting laser light of different wavelengths, and an objective lens that irradiates the sample with laser light emitted from the laser light source,
A beam splitter that is arranged between the laser light source and the objective lens and branches a part of the laser light from the laser light source; a light amount detecting means for detecting the light amount of the laser light branched by the beam splitter; A laser microscope comprising: a diaphragm device disposed between the light amount detection means and the beam splitter, for limiting a beam diameter.   The laser microscope according to claim 1, wherein the beam splitter is disposed between an imaging lens that forms an image of light collected by the objective lens and the objective lens.   The laser microscope according to claim 3, wherein the aperture device is disposed at a position optically equivalent to a pupil position of the objective lens.   The laser microscope according to any one of claims 1 to 4, wherein the beam splitter is provided so as to be inserted into and removed from an optical path between the laser light source and the objective lens.   The laser microscope according to claim 1, wherein the aperture device has an aperture shape substantially the same as an aperture shape of a pupil position of the objective lens.   2. The laser microscope according to claim 1, wherein the aperture diameter of the aperture stop device is a dimension obtained by multiplying the magnification of an optical system arranged between the objective lens and the beam splitter by the aperture diameter of the pupil position of the objective lens. .   The laser microscope according to claim 1, wherein the aperture device is provided so that its opening diameter can be changed. The objective lens is provided interchangeably,
The laser microscope according to claim 8, further comprising an adjusting device that adjusts an aperture diameter of the aperture device in accordance with an aperture diameter of a pupil position of the objective lens arranged on an optical axis.

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