JP2006220953A - Laser beam emitter and microscope equipment with laser beam emitter - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a laser beam emitter capable of variably forming both of a point light source and a surface light source with a simple constitution and to provide microscope equipment with the laser beam emitter. <P>SOLUTION: The laser beam emitter is provided with: a lens 9 in which an I-O distance as an interval between an object point 11 and an image point 10 is defined in a fixed value; an optical fiber 1 which guides a laser beam from a light emission point 12 as a point light source; a luminous flux diaphragm 5 arranged on the object point 11 position of a lens 9; and an emission unit 13 switching a first state that forms the light emission point 12 or a projection image of the light emission point 12 on the position of the luminous flux diaphragm 5 and a second state that applies light from the light emission point 12 so as to satisfy a diameter of the luminous flux diaphragm 5. In the first state, the light emission point 12 is projected on the image point 10 position of the lens 9 and, in the second state, a shape of the luminous flux diaphragm 5 is projected on the image point 10 position. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a laser light irradiation apparatus that guides laser light using an optical fiber and irradiates an observation sample with the guided laser light, and a microscope apparatus with a laser light irradiation apparatus.

  Conventionally, a substance that reacts only to light of a specific wavelength (stimulation light), living cells, fluorescent specimens, etc. is irradiated directly or under a microscope with a stimulus light to observe the reaction of a specific part. And measuring changes are widely performed. In general, for such an irradiation apparatus, (1) irradiating a small area of a certain area with a uniform amount of light (surface irradiation or spot irradiation), and (2) irradiating a minute portion with high intensity light. (Point irradiation) has two different purposes. Hereinafter, the one corresponding to (1) is referred to as surface irradiation, and the one corresponding to (2) is referred to as point irradiation.

  As an apparatus that performs surface irradiation, for example, an irradiation apparatus using a fiber bundle and a diffuser using a halogen, xenon lamp, mercury lamp, or the like as a light source is known. For the point irradiation, a laser light source and a single fiber (single-mode fiber or multi-mode fiber) are used, and the output end of the fiber is regarded as a point light source, and the optical system is configured to project it onto the specimen. Thus, the minute part of the specimen is illuminated with high intensity.

  Also, guide light that does not cause stimulation to the specimen, such as red laser, is used to drop the spot light onto the specimen at a position where the specimen's stimulation light is to be applied, and then the stimulation light is applied to the center of the spot. Applications are frequently used in recent cell biology.

  Moreover, there exists patent document 1 regarding the technique of surface-irradiating a sample with the laser beam introduce | transduced with the optical fiber. In this method, the exit end of the optical fiber is placed at a position conjugate to the aperture stop of the illumination optical system of the microscope so that the surface of the specimen is evenly illuminated. The angle is adjustable.

JP 2004-38139 A

  However, the conventional laser beam irradiation apparatus cannot combine point irradiation and surface irradiation. For example, in a surface irradiation device using a fiber bundle (fiber bundle), it is difficult in principle to perform minute spot irradiation because the exit end of the fiber bundle has a large area. It became. In patent document 1 which performs surface irradiation using a laser light source and a single fiber, nothing is described regarding combined use with point irradiation.

  The present invention has been made in view of the above, and an object of the present invention is to provide a laser light irradiation apparatus and a microscope apparatus with a laser light irradiation apparatus that can easily switch between point irradiation and surface irradiation with a simple configuration. To do.

  In order to achieve the above object, a laser beam irradiation apparatus according to claim 1 includes an irradiation lens system in which an IO distance, which is an interval between an object point and an image point, is set to a fixed value, and a laser beam from a light source. A light guide means for guiding the light source as a point light source, a light beam stop disposed at an object point position of the irradiation lens system, and a first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop; An emission unit that can switch between a second state of irradiation so as to satisfy the light beam aperture diameter by light from the point light source, and in the first state, at the image point position of the irradiation lens system A point light source is projected, and the shape of the beam stop is projected at the image point position in the second state.

  According to a second aspect of the present invention, there is provided the laser light irradiation apparatus according to the above invention, wherein the emission unit includes the first state in which the point light source is positioned at the object point position of the irradiation lens system, and the point light source. Switching means for switching to a second state in which the point light source is positioned at a position where the divergent light beam satisfies the light beam aperture diameter is provided.

  According to a third aspect of the present invention, there is provided the laser beam irradiation apparatus according to the above invention, wherein the emission unit converts a divergent light beam from the point light source into a parallel light beam having a diameter larger than an effective diameter of the light beam stop; A condensing lens for condensing the collimated light beam from the collimating lens at the object point position of the irradiation lens system, the first state in which the condensing lens is inserted into the optical axis, and an outer side of the optical axis. Switching means for switching to the second state.

  According to a fourth aspect of the present invention, there is provided the laser beam irradiation apparatus according to the above invention, wherein an optical path branching unit is provided between the irradiation lens system and the light beam stop, and the irradiation lens system is provided on the branched optical path. A second light beam stop disposed at the point position and a second exit unit for the second light beam stop are provided.

  According to a fifth aspect of the present invention, there is provided the laser beam irradiation apparatus according to the above invention, wherein the irradiation position is adjusted with respect to the irradiation lens system by integrally moving the beam stop and the emission unit within a plane perpendicular to the optical axis. Means are provided.

  According to a sixth aspect of the present invention, there is provided a laser beam irradiation apparatus according to the above invention, further comprising aperture variable means for varying the shape or size of the light beam aperture.

  According to a seventh aspect of the present invention, in the above-described invention, the irradiation lens system is an infinity correction optical system, and the lens on the image point side can be replaced with a lens having a different focal length. Features.

  In the laser beam irradiation apparatus according to claim 8, in the above invention, the irradiation system lens has a configuration similar to a Koehler illumination optical system of a microscope, and the light source position in the first state is a field stop of the microscope. The light source position in the second state is the brightness stop position of the microscope.

  According to a ninth aspect of the present invention, in the laser light irradiation apparatus according to the above invention, the light source includes a shutter, a wavelength selection unit, and a light amount adjustment unit.

  According to a tenth aspect of the present invention, there is provided the laser light irradiation apparatus according to the above invention, wherein the light guide means includes an optical fiber that guides the laser light, and uses an emission end of the optical fiber as the point light source. And

  The laser beam irradiation apparatus according to an eleventh aspect is characterized in that, in the above invention, the optical fiber is a single mode fiber.

  In the laser beam irradiation apparatus according to a twelfth aspect of the present invention, in the above invention, the laser light source oscillates two types of laser beams having different wavelengths, and the light guide unit individually divides the two types of laser beams. And two optical fibers for individually guiding laser beams to the two emission units.

  According to a thirteenth aspect of the present invention, there is provided a microscope apparatus with a laser beam irradiation apparatus according to the above invention, wherein an object point, an image point, An irradiation lens system in which the I-O distance, which is an interval, is fixed, a light guide means for guiding laser light from the light source as a point light source, and a beam stop disposed at an object point position of the irradiation lens system A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop, and a second state in which the light beam stop is irradiated with the light from the point light source so as to satisfy the effective diameter. The point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the light beam stop at the image point position in the second state. Projected A microscope apparatus with a laser light irradiation apparatus incorporating a light irradiation apparatus, comprising: an optical path branching unit that branches an optical path of the Koehler illumination device; and the light beam stop and the emission unit on the optical path branched by the optical path branching unit The position of the light beam stop is a position conjugate with the field stop of the Koehler illumination device, and the point light source position in the second state is a position satisfying the light beam stop diameter. The objective lens and the projection lens are used as the part.

  A microscope apparatus with a laser beam irradiation apparatus according to a fourteenth aspect is the above invention, wherein the emission unit positions the point light source at the object point position of the irradiation lens system, and the point. Switching means for switching to a second state in which the point light source is positioned at a position where the divergent light beam from the light source sufficiently satisfies the effective diameter of the light beam stop is provided.

  In the microscope apparatus with a laser beam irradiation apparatus according to claim 15, in the above invention, the emission unit converts a divergent light beam from the point light source into a parallel light beam having a diameter larger than an effective diameter of the light beam stop. A collimating lens, a condensing lens for condensing the parallel light flux from the collimating lens at the object point position of the irradiation lens system, the first state in which the condensing lens is inserted in the optical axis, and the light Switching means for switching to the second state removed from the shaft.

  According to a sixteenth aspect of the present invention, in the microscope apparatus with a laser beam irradiation apparatus according to the above invention, an optical path extension lens is provided on the optical path branched by the optical path branching means.

  According to a seventeenth aspect of the present invention, there is provided a microscope apparatus with a laser beam irradiation apparatus according to the above invention, wherein an object point and an image point are added to an upright or inverted microscope apparatus having an objective lens, a specimen mounting stage, and an observation optical path. An irradiation lens system in which the I-O distance, which is an interval, is set to a fixed value; a light guide unit that guides laser light from the light source as a point light source; and a beam stop disposed at an object point position of the irradiation lens system; A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop, and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy an effective diameter thereof. The point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the beam stop is formed at the image point position in the second state. Projected laser light irradiation A microscope apparatus with a laser beam irradiation apparatus incorporating a device, provided with an optical path branching means for branching the observation optical path, and arranging the light beam stop and the emission unit on the optical path branched by the optical path branching means, The position of the beam stop is a conjugate position with the imaging position of the observation optical path, and the objective lens and the imaging lens are used as a part of the irradiation lens system.

  The microscope apparatus with a laser beam irradiation apparatus according to claim 18 is the above-described invention, wherein the emission unit positions the point light source at the object point position of the irradiation lens system; Switching means for switching to a second state in which the point light source is positioned at a position where the divergent light beam from the light source sufficiently satisfies the effective diameter of the light beam stop is provided.

  In the microscope apparatus with a laser beam irradiation apparatus according to claim 19, in the above invention, the emission unit converts a divergent light beam from the point light source into a parallel light beam having a diameter larger than an effective diameter of the light beam stop. A collimating lens, a condensing lens for condensing the parallel light flux from the collimating lens at the object point position of the irradiation lens system, the first state in which the condensing lens is inserted in the optical axis, and the light Switching means for switching to the second state removed from the shaft.

  A microscope apparatus with a laser beam irradiation apparatus according to a twentieth aspect of the present invention is the inverted microscope apparatus having the objective lens, the sample stage, and the transmission illumination device according to the above invention, wherein I is the distance between the object point and the image point. An irradiation lens system in which the -O distance is set to a fixed value; a light guide means for guiding laser light from the light source as a point light source; a light beam stop disposed at an object point position of the irradiation lens system; Switchable between a first state in which the point light source or a projected image of the point light source is formed at a position and a second state in which the beam stop is irradiated with light from the point light source so as to satisfy its effective diameter. An emission unit, and in the first state, the point light source is projected onto the image point position of the irradiation lens system, and in the second state, the shape of the beam stop is projected onto the image point position. Set up a laser beam irradiation device A microscope apparatus with a laser beam irradiation device, wherein an optical path branching means is provided between the specimen stage and the condenser lens of the transmission illumination device, and the irradiation lens system is on the optical path branched by the optical path branching means And the light beam stop and the emission unit, and the image point position of the irradiation lens system coincides with the focus position of the objective lens.

  Furthermore, in the microscope device with a laser beam irradiation apparatus according to claim 21, in the above invention, the emission unit positions the point light source at the object point position of the irradiation lens system, and the point. Switching means for switching to a second state in which the point light source is positioned at a position where the divergent light beam from the light source sufficiently satisfies the effective diameter of the light beam stop is provided.

  In the microscope apparatus with a laser beam irradiation apparatus according to a twenty-second aspect, in the above invention, the emission unit converts a divergent light beam from the point light source into a parallel light beam having a diameter larger than an effective diameter of the light beam stop. A collimating lens, a condensing lens for condensing the parallel light flux from the collimating lens at the object point position of the irradiation lens system, the first state in which the condensing lens is inserted in the optical axis, and the light Switching means for switching to the second state removed from the shaft.

  According to a twenty-third aspect of the present invention, there is provided the microscope apparatus with a laser beam irradiation apparatus according to the above-described invention, an imaging unit that images an object, a display unit that displays the captured observation image, and the observation image on the display unit. Position designation means for designating an arbitrary point, irradiation position adjustment means for integrally moving the light beam stop and the emission unit in the XY directions perpendicular to the optical axis with respect to the irradiation lens system, and laser light Wavelength switching means for switching the wavelength between the stimulation light to the specimen and the guide light that does not react with the specimen, the type of laser light irradiated to the specimen, one of the stimulation light or the guide light, the point light source image or the light beam Irradiation light setting means set as a combination with one of the images of the aperture, and the wavelength switching means and the switching unit of the emission unit based on the type of irradiation light set by the wavelength switching means Gosuru with and control means for controlling the irradiation position adjusting unit based on the position specified by the position specifying means, and then irradiating the laser beam set to the specified position type.

  According to a twenty-fourth aspect of the present invention, in the microscope apparatus with a laser beam irradiation device according to the above invention, the microscope device further includes a light control unit that adjusts a light amount of the laser beam, and the irradiation light setting unit controls the light amount of the laser beam to be irradiated. It is possible to set, and the control means controls the light control means based on the set amount of laser light.

  According to a 25th aspect of the present invention, there is provided a microscope apparatus with a laser beam irradiation apparatus according to the above-mentioned invention, wherein a stereoscopic microscope apparatus having a TV optical path on one of the two observation optical axes is provided at an interval between an object point and an image point. An irradiation lens system in which a certain I-O distance is set to a fixed value, light guide means for guiding laser light from the light source as a point light source, a light beam stop disposed at an object point position of the irradiation lens system, and the light beam Switching between a first state in which the point light source or a projected image of the point light source is formed at the position of the stop and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy its effective diameter. The point light source is projected onto the image point position of the irradiation lens system in the first state, and the shape of the beam stop is projected onto the image point position in the second state. Built-in laser beam irradiation device This is a stereomicroscope apparatus with a laser beam irradiation device, wherein an optical path branching means is provided between the objective lens and the sample and on the side where the TV optical path is not provided, and branched by the optical path branching means The irradiation lens system, the light beam stop, and the emission unit are disposed on an optical path, and an image point position of the irradiation lens system coincides with a focus position of the objective lens.

  A microscope apparatus with a laser beam irradiation apparatus according to a twenty-sixth aspect is the above invention, wherein the emission unit positions the point light source at the object point position of the irradiation lens system; Switching means for switching to a second state in which the point light source is positioned at a position where the divergent light beam from the light source sufficiently satisfies the effective diameter of the light beam stop is provided.

  In the microscope apparatus with a laser beam irradiation apparatus according to a twenty-seventh aspect, in the above invention, the emission unit converts a divergent light beam from the point light source into a parallel light beam having a diameter larger than an effective diameter of the light beam stop. A collimating lens, a condensing lens for condensing the parallel light flux from the collimating lens at the object point position of the irradiation lens system, the first state in which the condensing lens is inserted in the optical axis, and the light Switching means for switching to the second state removed from the shaft.

  According to a 28th aspect of the present invention, in the microscope apparatus with a laser beam irradiation apparatus, in the above invention, an imaging unit that images an object, a display unit that displays the captured observation image, and the observation image on the display unit Position designation means for designating an arbitrary point, irradiation position adjustment means for integrally moving the light beam stop and the emission unit in the XY directions perpendicular to the optical axis with respect to the irradiation lens system, and laser light The wavelength is set by the irradiation light setting means for setting the wavelength as a combination of one of the stimulation light to the specimen and the guide light to which the specimen does not react, and one of the point light source image or the image of the beam stop, and the wavelength switching means. The wavelength switching means and the emission unit switching means are controlled based on the type of irradiated light and the irradiation position adjusting means is controlled based on the position designated by the position designation means. And control means for, and then irradiating the laser beam set to the specified position type.

  A microscope apparatus with a laser beam irradiation apparatus according to a twenty-ninth aspect of the present invention is the above invention, further comprising a light control unit that adjusts a light amount of the laser beam, and the irradiation light setting unit controls the light amount of the laser beam to be irradiated. It is possible to set, and the control means controls the light control means based on the set amount of laser light.

  Throughout the present invention, a single mode fiber (SMF) is preferable as the light guiding means for guiding the laser light to the irradiation device in that an ideal point light source can be obtained. However, it is also within the scope of the present invention to use a multimode fiber (MMF) when a strict point source is not required.

  The laser light irradiation apparatus and the microscope apparatus with a laser light irradiation apparatus according to the present invention can variably form a point light source and a surface light source with a simple configuration, and one laser light source can be used as both a point light source and a surface light source. There is an effect.

  Exemplary embodiments of a laser beam irradiation apparatus and a microscope apparatus with a laser beam irradiation apparatus according to the present invention will be explained below in detail with reference to the accompanying drawings.

(Embodiment 1)
First, a laser beam irradiation apparatus according to the present invention will be described. 1 and 2 are block diagrams showing a schematic configuration of the laser beam irradiation apparatus according to the first embodiment. FIG. 1 shows a case where the laser light irradiation apparatus 1 forms a point light source and laser light irradiates the image point 16. FIG. 2 shows a case where the laser light irradiation apparatus 1 forms a surface light source and the laser light Is a case where the image point 16 is surface-irradiated.

  First, with reference to FIG. 1, the 1st state in which this laser beam irradiation apparatus 1 forms a point light source is demonstrated. In FIG. 1, the laser light irradiation apparatus 1 includes an irradiation unit 10 and an irradiation lens 11, and the irradiation unit 10 and the irradiation lens 11 are connected and fixed via a housing 14. The irradiation unit 10 includes an emission unit 10a and a beam stop 10b. The emission unit 10a holds the optical fiber 12 that guides the laser light, and moves along the optical axis direction of the laser light. A slider 13 for fixing, a fixing screw 18 for fixing the position of the slider 13, and a stop plate 15. The light beam stop 10b blocks the laser light that has expanded beyond the effective diameter of the light beam stop 10b, and restricts the light beam of the laser light. When the slider 13 moves, the tip of the optical fiber 12 moves from the position of the light beam stop 10b to a position where the spread of the laser light emitted from the optical fiber 12 becomes equal to or larger than the effective diameter of the light beam stop.

  The irradiation lens 11 includes a lens 19, a lens frame 17 that holds the lens 19, and a presser ring 20 that fixes the lens 19 together with the lens frame 17. The object point 22 and the image point 16 are in a conjugate positional relationship with respect to the lens 19, and the object point 22 is at the position of the beam stop 10b. The distance between the object point 22 and the image point 16 is the IO distance of the irradiation lens 19, and this IO distance is a fixed value. The distance from the tip of the lens frame 17 to the image point 16 is WD (working distance). Further, the light emitting point 21 which is the laser light emitting end of the optical fiber 12 can be regarded as a point light source when the optical fiber 12 is SMF.

  When the tip of the optical fiber 12 is at the position of the beam stop 10b, the position of the light emitting point 21 and the position of the object point 22 coincide. Here, since the light emitting point 21 is a point light source, the laser light emitted from the light emitting point 21 is not limited by the light beam stop by the light beam stop 10b. For this reason, the laser beam emitted from the light emitting point 21 converges on the image point 16 and forms a point image of the light emitting point 21 at the image point 16. When an observation sample or the like is placed at the image point 16 in this state, the laser beam emitted from the light emitting point 21 is irradiated to the observation sample or the like.

  Next, a second state in which the laser light irradiation apparatus 1 forms a surface light source will be described with reference to FIG. As shown in FIG. 2, when the fixing screw 18 is loosened and the rear end of the slider 13 is moved to the position of the stop plate 15, the light emitting point 21 of the optical fiber 12 moves together with the slider 13 to form the second state. To do. When laser light is emitted from the optical fiber 12 in this second state, the laser light emitted from the light emitting point 21 has a predetermined divergence angle. The expanded laser light is narrowed down to the effective diameter of the light beam stop 10b by the light beam stop 10b. The laser beam focused by the beam stop 10 b enters the lens 19, converges at the condensing point 23, and forms a surface image (spot) obtained by projecting the hole shape of the beam stop 10 b at the image point 16 corresponding to the object point 22. Form an image. The surface image formed at the image point 16 is a hole-shaped image of the beam stop 10b, and thus has a clear outline. As a result, when the observation sample is arranged at the image point 16 in the second state, the surface irradiation is performed with the laser beam that projects the hole of the beam stop 10b on the observation sample.

  That is, the laser beam irradiation apparatus 1 performs point irradiation on the image point 16 by forming the first state at the same position, and the image point 16 by forming the second state. Surface irradiation can be performed.

  Here, since the flexible optical fiber 12 is used as the laser light guiding means, the light emitting point 21 can be easily moved, and the point light source and the surface light source are easily formed at the position of the beam stop 10b. it can.

  Note that the light amount distribution with respect to the numerical aperture (NA) of the laser light at the light emitting point 21 in the second state is a Gaussian distribution with respect to NA as shown in FIG. When a surface light source is formed with an NA in which the nominal NA obtained by cutting a portion where the amount of light is 0 to 13% is reduced to about 1/3, a portion with a more even light amount distribution can be taken out and the uniformity of the light amount distribution can be improved. it can. Actually, when an optical fiber having an NA of 0.1 to 0.15 at the light emitting point 21 is used and approximately 1/3 of the NA is used, when the aperture diameter of the beam stop 10b is 1 mm, the beam stop 10b The distance to the light emitting point 21 is about 5 mm.

  In the first embodiment, the point light source and the surface light source are varied by moving the slider 13 manually. However, the slider 13 is driven by a motor or the like, and the drive control of the motor or the like is performed. May be performed.

  The laser beam irradiation apparatus 1 shown in the first embodiment guides a laser beam by using an optical fiber 12 and varies the distance between the light beam stop 10b and the light emitting point 21 of the optical fiber 12 to thereby change the image point. The point irradiation and the surface irradiation with respect to 16 can be easily realized.

  In addition, since two color lasers can be switched and introduced into the optical fiber 12, the laser of one color can be irradiated as a surface light source, and the other can be irradiated as a point light source by moving the light emitting point 21.

(Embodiment 2)
Next, a laser beam irradiation apparatus according to Embodiment 2 of the present invention will be described. In the first embodiment, the point light source and the surface light source are variably formed at the position of the light beam stop 10b by moving the slider 13, but in this second embodiment, the optical fiber and the light beam stop A collimating lens and a condensing lens are arranged between them, and a point light source and a surface light source are variably formed at the position of the beam stop by inserting and removing the condensing lens.

  4 and 5 are block diagrams showing a schematic configuration of the laser beam irradiation apparatus 2 according to the second embodiment. FIG. 4 shows a case where the laser irradiation device 2 forms a point light source in the first state at the position of the beam stop 25b, and FIG. 5 shows that the laser irradiation device 2 is positioned at the position of the beam stop 25b. The case where the surface light source which is a state is formed is shown.

  In FIG. 4, the laser beam irradiation apparatus 2 includes an irradiation unit 25 and an irradiation lens 26, and the irradiation unit 25 includes an emission unit 25a and a beam stop 25b. The emission unit 25a includes an optical fiber 27 that guides the laser light, a collimator lens 29 that converts the laser light emitted from the light emitting point 28 of the optical fiber 27 into parallel light, and a beam stop for the laser light converted into parallel light. And a condensing lens 31 that condenses the object point 37 located at the aperture position 25b. The condenser lens 31 is housed in an insertion / removal unit 32, and the insertion / removal unit 32 can be inserted into and removed from the injection unit 25a. The insertion / removal of the insertion / removal unit 32 is converted by the collimator lens 29. This means insertion / removal of the condenser lens 31 with respect to the parallel light flux.

  The irradiation lens 26 includes a relay lens 34 and a light projection lens 35. The relay lens 34 converts laser light emitted from the object point 37 into parallel light, and the light projection lens 35 is converted into parallel light. The laser beam is focused on the image point 36. In this case, the object point 37 and the image point 36 which are the aperture position of the light beam aperture 25b are in a conjugate positional relationship with each other.

  When the insertion / removal unit 32 is inserted, the laser light emitted from the light emitting point 28 of the optical fiber 27 is converted into parallel light by the collimator lens 29, and this parallel light is converted by the condenser lens 31 into the light beam stop 25 b. The light is condensed at the stop position, and the light emission point 28 is projected onto the object point 37 at the stop position. The laser light that has passed through the beam stop 25 b is converted into parallel light by the relay lens 34 and further imaged at the image point 36 by the light projecting lens 35. As a result, the laser light emitted from the light emission point 28 converges on the image point 36, and a point image of the light emission point 28 is formed on the image point 36. Here, when an observation sample or the like is arranged at the image point 36, the laser light emitted from the light emitting point 28 is irradiated to the observation sample or the like, and the observation sample or the like is irradiated with a laser beam having high energy.

  On the other hand, as shown in FIG. 5, when the insertion / removal unit 32 is removed from the insertion position, the laser light emitted from the light emitting point 28 of the optical fiber 27 is converted into parallel light by the collimator lens 29, and as it is. The position 25b is reached. The laser beam that has reached the beam stop 25 b is focused by the aperture diameter of the beam stop 25 b, and the reduced beam enters the irradiation lens 26.

  The laser light incident on the irradiation lens 26 is condensed at a condensing point 38 by the relay lens 34 and the light projecting lens 35, and then the shape of the aperture of the light beam stop 25 b that is the object point 37 is projected onto the image point 36. The formed plane image is formed. Here, when an observation sample or the like is placed at the image point 36, surface irradiation of a laser beam with a clear outline is performed as in the surface irradiation described in the first embodiment.

  Here, an infinite correction optical system is formed between the collimating lens 29 and the condenser lens 31 and between the relay lens 34 and the light projecting lens 35 as shown in FIG. WD (working distance) can be changed by exchanging for a different focal length. In the second embodiment, since the point light source and the surface light source are variably formed only by inserting / removing the insertion / removal unit 32, the configuration can be further simplified.

  In the second embodiment, a collimating lens 29 and a condensing lens 31 are arranged between the optical fiber 27 and the light beam stop 25b, and the condensing lens 31 is inserted into the parallel light beam converted by the collimating lens 29. By removing, the point light source and the surface light source can be formed variably and easily at the stop position of the light beam stop 25b.

(Embodiment 3)
Next, a laser beam irradiation apparatus according to Embodiment 3 of the present invention will be described. In the first and second embodiments, one laser beam irradiation device variably forms a point light source and a surface light source. However, in the third embodiment, two laser beam irradiation devices are combined to be different. A laser beam having a wavelength is irradiated to one image point.

  FIG. 6 is a block diagram showing a schematic configuration of a laser beam irradiation apparatus according to Embodiment 3 of the present invention. In FIG. 6, the laser beam irradiation device 3 is configured to emit light from the irradiation unit 10 described in the first embodiment, the irradiation unit 25 illustrated in the second embodiment, and the laser beams emitted from the irradiation units 10 and 25. It has the dichroic mirror 39 that combines without reduction and the irradiation lens 26 shown in the second embodiment. Here, the object point 22 of the irradiation unit 10 and the object point 37 of the irradiation unit 25 have a common image point 40 via the dichroic mirror 39 and the irradiation lens 26.

  In this laser beam irradiation apparatus 3, for example, when a laser beam having a wavelength of 633 nm is emitted from the irradiation unit 10, a laser beam having a wavelength of 405 nm is emitted from the irradiation unit 25, and an observation sample is placed at the image point 40, the observation sample The same part can be surface-irradiated or spot-irradiated with laser light having different wavelengths.

  Here, when SMF is used for the light guide fiber, SMF has good transmission characteristics for laser light of a predetermined wavelength, but has poor transmission characteristics for laser light of other wavelengths. When irradiating the same point on the observation sample with laser light having different wavelengths (633 nm, 405 nm), it is desirable to use SMF corresponding to each wavelength.

  In the third embodiment, the irradiation unit 10 and the irradiation unit 25 are combined. However, the configuration is not limited to this, and the two irradiation units 10 or the two irradiation units 25 may be combined. Furthermore, the wavelength of the laser light guided to each irradiation unit 10, 25 is also arbitrary.

  In the third embodiment, the irradiation units 10 and 25 are combined, and laser beams having different wavelengths are synthesized using the dichroic mirror 39, whereby point irradiation and surface irradiation are individually performed on the same part on the observation sample. Can be done. For example, the surface of the sample is positioned by irradiating the surface with 633 nm red laser light, the point of 405 nm is irradiated at the same position, and the position of the light stimulus is determined by the easily observable red surface irradiation light. Light stimulation can be applied to the center point of the part at 405 nm.

(Embodiment 4)
Next, a laser beam irradiation apparatus according to Embodiment 4 of the present invention will be described. In the first to third embodiments described above, the optical axis of the laser light emitted from the laser light irradiation devices 1 to 3 is fixed. However, in this fourth embodiment, XY is provided between the irradiation unit and the irradiation lens. A movable mechanism is provided so that the optical axis of the laser beam can be varied on the observation sample.

  7 and 8 are block diagrams showing a schematic configuration of a laser beam irradiation apparatus according to Embodiment 4 of the present invention. As shown in FIG. 7, in the laser beam irradiation device 4, the irradiation unit 10 and the irradiation lens 11 are connected via an XY movable mechanism 41.

  Here, the XY movable mechanism 41 is movable in a direction perpendicular to the optical axis of the laser light emitted from the irradiation unit 10. When the XY movable mechanism 41 is moved, the XY movable mechanism 41 is conjugate with the object point of the irradiation unit 10. The position of the image point 16 moves in conjunction. For example, when the observation sample is arranged at the image point 16 using the irradiation unit 10, the image point 16 also moves on the observation sample in conjunction with the movement of the XY movable mechanism 41. As a result, the irradiation surface of the laser light can be moved only by moving the XY movable mechanism 41 without moving the observation sample. The XY movable mechanism 41 can be realized using, for example, the B27-100 series XY movable mechanism manufactured by Suruga Seiki Co., Ltd.

  As a specific application, when an electrode is implanted into a sample such as a nerve cell, and a stimulation light is irradiated on a specific part of the sample or the entire sample at regular intervals, the state of nerve connection is known by the current generated at the electrode. When the stage is moved in this case, it is used to prevent the electrodes from being detached, or to irradiate stimulation light at an arbitrary position in the screen imaged by the CCD camera and store the state as an image.

  Further, as shown in FIG. 8, a laser beam irradiation device 5 in which the irradiation unit 25 and the irradiation lens 26 described in the second embodiment are connected via an XY movable mechanism 41 may be used. In the case of this laser beam irradiation apparatus 5 as well, it is possible to move the illumination part of the observation sample arranged at the image point 36 or the part to be stimulated only by moving the XY movable mechanism 41 without moving the observation sample.

(Embodiment 5)
Next, a laser beam irradiation apparatus according to Embodiment 5 of the present invention will be described. In the first to fourth embodiments described above, the light beam stops 10b and 25b of the laser light irradiation devices 1 to 4 are fixed. However, in the fifth embodiment, the shape of the diaphragm of the light beam stop can be varied.

  FIG. 9 is a block diagram showing a schematic configuration of a laser beam irradiation apparatus according to Embodiment 5 of the present invention. As shown in FIG. 9, the laser beam irradiation apparatus 1A includes a revolver 10A instead of the light beam stop 10b described in the first embodiment. Other configurations are the same as those of the laser beam irradiation apparatus 1 shown in the first embodiment, and the same components are denoted by the same reference numerals.

  A plurality of beam stops 10b, 10c,... Having different sizes and shapes are incorporated in the revolver 10A. By rotating the revolver 10A, a desired beam stop is provided between the emission unit 10a and the housing 14. It is designed to be incorporated.

  Here, when the laser light irradiation apparatus 1A forms a surface light source and the shape of the surface image at the image point 16 and the NA are varied, a desired beam stop is placed between the emission unit 10a and the housing 14. By incorporating it, a plane image having a desired NA and shape can be formed at the image point 16.

  In the fifth embodiment, the revolver 10A is incorporated in the laser beam irradiation apparatus 1 described in the first embodiment. However, the laser beam irradiation apparatus 2 in the second embodiment is not limited to this. You may make it incorporate in the laser beam irradiation apparatus of embodiment. In the fifth embodiment, the rotary revolver 10A is used. However, a linear slider may be used, or individual beam stops 10b, 10c,... A beam stop with a variable aperture shape may be provided. Furthermore, these light beam stops may be changed automatically instead of manually.

  In the fifth embodiment, a desired plane image is obtained by providing a revolver 10A incorporating a plurality of light beam stops 10b, 10c.

(Embodiment 6)
Next, a laser beam irradiation apparatus according to Embodiment 6 of the present invention will be described. In the first to fifth embodiments described above, the positions of the image points 16 and 36 of the laser light irradiation devices 1 to 5 are fixed in the optical axis direction of the laser light. In the sixth embodiment, the position of the image point. Can be varied in the optical axis direction.

  FIG. 10 is a block diagram showing a schematic configuration of a laser beam irradiation apparatus according to Embodiment 6 of the present invention. As shown in FIG. 10, this laser beam irradiation apparatus 2A is provided with an irradiation lens 26A in place of the irradiation lens 26 shown in the second embodiment, and the irradiation lens 26A includes a relay lens 34 and a light projection lens unit 35A. Have. The projection lens unit 35A has a plurality of projection lenses 35a and 35b. Other configurations are the same as those of the laser beam irradiation apparatus 2 shown in the second embodiment, and the same components are denoted by the same reference numerals.

  As shown in FIG. 10, the projection lens unit 35A has a plurality of projection lenses 35a and 35b having different focal lengths, and the projection lenses 35a and 35b are exchanged by inserting and removing the projection lens unit 35A. Thus, it is incorporated into the irradiation lens 26A. For example, when the light projection lens 35a is incorporated, the image point 36a is far from the irradiation lens 26A, but when the light projection lens 35b is incorporated, the image point 36b is near the irradiation lens 26A. .

  In the sixth embodiment, the positions of the image points 36a and 36b in the optical axis direction of the laser light emitted from the irradiation unit 25 can be varied by exchanging the projection lenses 35a and 35b having different focal lengths. . In addition, you may make it replace this projection lens unit 35A using a revolver.

(Embodiment 7)
Next, a seventh embodiment according to the present invention will be described. In the seventh embodiment, the light projecting optical system is made the same as the Koehler illumination optical system of a microscope, and the laser light emitted from the laser light irradiation device performs point irradiation at the image point in the first state, and in the second state. The light source is placed at the aperture stop position of the Koehler illumination optical system so that Koehler irradiation is performed. Since the light source used here is a point light source, it is optically “collimated irradiation”. However, the optical system has a configuration of Koehler illumination of a microscope, and when the apparatus of this embodiment is incorporated in a microscope, Since the Koehler illumination optical system can be used, it will be referred to as “Kohler irradiation” in the present invention.

  It should be noted that the point light source in the second state does not necessarily need to be accurately at the brightness stop, and may only satisfy the light beam stop of the irradiation device.

  11 and 12 are block diagrams showing a schematic configuration of a laser beam irradiation apparatus according to Embodiment 7 of the present invention. FIG. 11 shows a case where the laser light emitted from the laser light irradiation device 6 becomes Koehler illumination at the image point 66, and FIG. 12 shows the laser light emitted from the laser light irradiation device 6 at the image point 66. The case where point irradiation is performed is shown. That is, the light emitting point 21 is at the brightness stop position of the microscope when Koehler irradiation is performed, and is at the field stop position of the microscope at the time of spot irradiation.

  As shown in the example of the lens arrangement in FIG. 11, the laser light irradiation device 6 includes an irradiation unit 10 having an emission unit 10 a and a light beam stop 10 b, a relay lens 63, and a light projection lens (condenser lens in a microscope) 65. And have. The relay lens 63 and the light projection lens 65 function as an irradiation lens. The light beam stop 10b is disposed at the position of the focal point f2 of the relay lens 63, and the light emitting point 21 and the condensing point 64 placed at the focal length 2f2 that is twice that of the lens 63 are conjugate to each other with respect to the relay lens 63. There is a relationship. The condensing point 64 is at the position of a focal point f1 formed on the light beam stop 10b side of the light projecting lens 65, and the image point 66 is a focal point f1 formed on the opposite side of the light projection lens 65 with respect to the light beam stop 10b. In the position.

  In this case, the laser light emitted from the light emitting point 21 which is a point light source forms an image as a point image on the condensing point 64. On the other hand, since the condensing point 64 is located at the focal point of the light projecting lens 65 on the light beam stop 10b side, the image point 66 which is the focal point of the light projecting lens 65 on the opposite side of the light beam stop 10b is The imaged point image is projected as parallel light, and at the same time, the edge 10 c of the light beam stop 10 b forms an image on the edge 10 c ′ of the image point 66.

  Here, when the observation sample is arranged at the image point 66, the laser light with respect to the observation sample becomes Koehler irradiation, and the observation sample is subjected to surface illumination that is more uniform than in the second embodiment. Since the laser light applied to the observation sample is approximated to parallel light, even if the observation sample is arranged slightly shifted from the image point 66 in the optical axis direction, illumination close to ideal is performed.

  Next, as shown in FIG. 12, when the light emission point 21 is moved to the stop position of the light beam stop 10b, the laser light emitted from the light emission point 21 forms an image at the image point 66 as a point image. Here, when the observation sample is arranged at the image point 66 and the laser beam is emitted, the laser beam is point-irradiated at the image point 66, and the observation sample can be stimulated.

(Modification)
Next, a modification of the seventh embodiment of the present invention will be described. In the above-described seventh embodiment, the irradiation unit 10 shown in the first embodiment is used so that the laser beam is Koehler irradiation at the image point 66. In this modification, the irradiation unit 10 shown in the second embodiment is used. The irradiation unit 25 is used so that the laser beam becomes Koehler irradiation at the image point.

  13 and 14 are block diagrams showing a schematic configuration of a laser beam irradiation apparatus which is a modification of the seventh embodiment of the present invention. FIG. 13 shows a case where the laser light emitted from the laser light irradiation device 7 becomes Koehler irradiation at the image point 78, and FIG. 14 shows the laser light emitted from the laser light irradiation device 7 at the image point 78. On the other hand, the case of point irradiation is shown.

  As shown in FIG. 13, the laser beam irradiation device 7 includes an irradiation unit 25 having an emission unit 25 a and a beam stop 25 b, a relay lens 75, and a light projecting lens (a condenser lens in a microscope) 77. The beam stop 25b is disposed at the focal position on the irradiation unit 25 side of the relay lens 75, and the focal position 76 on the opposite side of the relay lens 75 with respect to the irradiation unit 25 is the focal position on the irradiation unit 25 side of the light projecting lens 77. . Further, the insertion / removal unit 32 is removed from the optical path of the laser beam.

  In this case, the laser light emitted from the light emitting point 28 becomes parallel light and enters the relay lens 75 via the light beam stop 25 b and is condensed at the focal position 76. The laser beam condensed at the focal position 76 diverges and enters the light projecting lens 77 and is converted into parallel light. With this parallel light, the hole shape of the beam stop 25b is imaged at the image point 78.

  Next, as shown in FIG. 14, when the insertion / removal unit 32 is inserted between the beam stop 25b and the collimating lens 29, the laser light emitted from the light emitting point 28 is condensed at the stop position of the beam stop 25b. Further, the light is condensed on the image point 78 via the relay lens 75 and the condenser lens 77, and a point image is formed on the image point 78. By such insertion / removal of the insertion / removal unit 32, the laser beam variably performs Kohler surface irradiation and point irradiation at the image point 78.

(Embodiment 8)
Next, a laser beam irradiation apparatus according to Embodiment 8 of the present invention will be described. In the eighth embodiment, the wavelength, intensity, and irradiation time of the laser light emitted from the laser light irradiation device are controlled.

  FIG. 15 is a block diagram showing a schematic configuration of a laser beam irradiation apparatus according to the eighth embodiment of the present invention. In FIG. 15, the laser beam irradiation apparatus 1B includes a bandpass filter 19a that selects a wavelength on the tip side of the irradiation lens 11 shown in the first embodiment, an ND filter 19b that controls the intensity of the laser beam, and a laser beam. And a shutter 19c that controls the irradiation time of the laser beam. Other configurations are the same as those of the laser beam irradiation apparatus 1 shown in the first embodiment, and the same components are denoted by the same reference numerals.

  As shown in FIG. 15, the laser emitted from the fiber guiding the multi-wavelength laser is emitted from the irradiation lens 11 toward the sample and is incident on the bandpass filter 19a, and only the laser light having a predetermined wavelength is emitted. pass. Further, the laser light incident on the ND filter 19b becomes laser light having a predetermined intensity. Further, the shutter 19c has a function of shielding the laser beam, and the irradiation time of the laser beam is controlled by opening and closing the shutter 19c. Therefore, by passing the laser light emitted from the irradiation lens 11 through the band-pass filter 19a and the ND filter 10b, it is possible to irradiate laser light having a desired wavelength and desired intensity, and furthermore, this laser light is opened / closed by the shutter 19c. The irradiation time can be controlled.

  In the eighth embodiment, the band-pass filter 19a, the ND filter 19b, and the shutter 19c are arranged at the laser beam emission side tip of the laser beam irradiation apparatus 1, but the laser beam irradiation shown in the second embodiment is performed. A band pass filter 19a, an ND filter 19b, and a shutter 19c may be disposed at the tip of the laser beam emission side of the apparatus 2. Further, a wavelength or intensity adjusting means may be provided on the laser light source side.

(Embodiment 9)
Next, a microscope apparatus with a laser light irradiation apparatus according to a ninth embodiment of the present invention will be described. FIG. 16 is a block diagram showing a schematic configuration of a microscope apparatus with a laser beam irradiation apparatus according to the ninth embodiment of the present invention. As shown in FIG. 16, the microscope apparatus 8 with a laser beam irradiation apparatus includes an irradiation unit 10 shown in the first embodiment, an irradiation unit 25 shown in the second embodiment, and an XY shown in the fourth embodiment. And a movable mechanism 41. An irradiation unit 10, 25, a dichroic mirror 82A, and a concave lens 81A for extending the optical path are fixed to the XY movable mechanism 41, and the optical axis of the laser light emitted from the irradiation unit 10, 25 is the dichroic mirror. 82A is synthesized.

  Further, this microscope apparatus 8 with a laser beam irradiation apparatus includes a mercury lamp 89 that illuminates an observation sample disposed at an image point 86, and a band that is emitted from the mercury lamp 89 and disposed at an arbitrary position on the lamp side from the field stop. A half mirror 88 that synthesizes the excitation light that has passed through the pass filter 19a and the laser light emitted from the irradiation units 10 and 25 and guides it toward the specimen, and the illumination light that is synthesized by the half mirror 88 is relayed to the objective lens. Relay lens 87, excitation light relayed by relay lens 87 and dichroic mirror 84 that reflects light from the emission unit, and these light reflected by dichroic mirror 84 are applied to image point 86 on the observation sample. The objective lens 85, the absorption filter 19c for extracting only the fluorescence emitted from the observation sample arranged at the image point 86, and the fluorescence C And a condensing lens 83 for condensing the D camera 80. The dichroic mirror 84 and the absorption filter 19e are attached to a mirror cube 84A that can be switched or exchanged.

  In this case, the image point 86 and the aperture position of the light beam stop of the irradiation unit 10 are conjugated with each other with respect to the optical system formed by the relay lens 87 and the objective lens 85, and the image point 86 and the irradiation unit 25. The aperture positions of the light beam apertures are also conjugated with each other.

  Here, an observation method when an observation sample in which Kaede fluorescent protein is expressed is arranged at the image point 86 will be described. Kaede is a protein that emits green fluorescence, but when irradiated with ultraviolet light, the fluorescence color irreversibly changes to red. For example, when Kaede is expressed in a neuron cell and irradiated with ultraviolet light aiming at one cell, it is possible to make only the extended projection out of the intricately intertwined neurites appear in red.

  The observation method starts with a mercury lamp 89, passes through a double-band excitation filter of 488 nm and 543 nm and a half mirror 88, and then is excited by a triple-band dichroic mirror unit that reflects 488 nm, 543 nm, and 405 nm. The light illuminates the observation sample placed at the image point 86 by the objective lens. At this time, green fluorescence is generated from the sample. Thereafter, a sample stage (not shown) on which the observation sample is placed is moved via the observation lens barrel 82, and the observation sample is roughly positioned. Next, a red laser (633 nm) is emitted from the emitting device 25, and an image of the light beam stop of the red irradiating device (a spot having a size about 1/20 of the observation field) is formed on the sample. The XY stage 41 of the irradiation apparatus is moved, and the red laser spot is moved to the position where the sample is to be irradiated while looking at the monitor of the CCD camera.

  Next, 405 nm stimulation light guided to the fiber of the irradiation device 10 is irradiated. Since the center of the red laser spot and the center of the 405 nm laser are set concentric in advance, the 405 nm laser is irradiated to the center of the spot. Then, the neuron at that site develops red color by the 543 nm excitation light that has followed the same path as the 488 nm excitation light, and over time, the entire cell connected to the neuron cell develops red color. This change over time can be observed from the CCD camera. Here, the reason why the 633 nm laser that has passed through the dichroic mirror 82A and reflected by the half mirror 88 and the triple band dichroic mirror reaches the sample is that the triple band dichroic mirror has a slight reflectivity with respect to 633 nm (general). A dichroic mirror has this character, and in the case of fluorescence observation, this red light amount is not significantly different from the fluorescence amount, which is convenient.

  Here, the half mirror 88 is described as a flat mirror, but a half mirror of a type in which two triangular prisms are joined is preferable. The reason is that, in the case of a flat plate half mirror, back surface reflection occurs and a red spot is formed on the sample, and the stimulus light point image is formed as a double image.

  The bandpass filter 19a is positioned on the lamp side of the half mirror 88 for the purpose of preventing the bandpass filter 19a from dimming the red laser or the stimulation light laser from the irradiation unit.

  In the ninth embodiment, by providing the irradiation units 10 and 25 and the XY movable mechanism 41 on which the irradiation units 10 and 25 are mounted, a laser beam having a wavelength of 633 nm emitted from the irradiation unit 25 is applied to the observation sample disposed at the image point 86. On the other hand, it can be used as a positioning index by surface irradiation, and the observation sample can be stimulated by the 405 nm laser light emitted from the irradiation unit 10.

  In the ninth embodiment, the laser light emitted from the irradiation unit 25 becomes surface irradiation at the image point 86, and the laser light emitted from the irradiation unit 10 becomes point irradiation and gives a stimulus to the observation sample. However, depending on the application, the irradiation unit 10 may emit a laser beam having a wavelength of 633 nm, and the irradiation unit 25 may emit a laser beam having a wavelength of 405 nm. Moreover, you may make it arrange | position two irradiation units 10 and 25 of any one of irradiation units 10 and 25. FIG. Further, although the mercury lamp 89 is used as the light source of the lamp light, a halogen lamp or the like may be used.

  In the ninth embodiment, the laser beam irradiation device is provided in the upright microscope apparatus, but is provided in the illumination side optical path of the inverted microscope apparatus or the observation side optical path of the inverted microscope apparatus. Also good.

(Modification)
Next, a modification of the ninth embodiment of the present invention will be described. In Embodiment 9 described above, the irradiation units 10 and 25 are both fixed, but in this modification, the irradiation units 10 and 25 are separated and placed in the imaging optical system and the projection optical system, respectively. ing.

  FIG. 17 is a block diagram showing a schematic configuration of a microscope apparatus with a laser beam irradiation apparatus, which is a modification of the ninth embodiment of the present invention. As shown in FIG. 17, this microscope apparatus 8A with a laser beam irradiation apparatus shows that the irradiation apparatus 10 or 25 can be attached to both a light projection tube and an imaging optical system, and can be appropriately selected according to the application. It is shown that. An XY movable mechanism 41 is fixed to the irradiation unit 25 so that the optical axis of the laser light emitted from the irradiation unit 25 moves on the observation sample. The irradiation unit 10 does not have the XY movable mechanism 41 and is configured to perform point irradiation and surface irradiation only at the center of the microscope field.

  As shown in FIG. 17, the half mirror 81 is disposed between the observation barrel 82 and the CCD camera 80, and the aperture position of the light beam stop of the irradiation unit 25 and the microscope image formation point 80 include the condenser lens 83 and the objective. The optical system formed by the lens 85 is conjugated with each other.

(Embodiment 10)
Next, a tenth embodiment of the present invention will be described. In Embodiment 9 described above, the laser light irradiation apparatus is attached to the upright microscope apparatus, but in Embodiment 10, the laser light irradiation apparatus is attached to the inverted microscope apparatus.

  FIG. 18 is a block diagram showing a schematic configuration of an inverted microscope apparatus with a laser beam irradiation apparatus according to the tenth embodiment of the present invention. As shown in FIG. 18, this inverted microscope apparatus 9 with a laser light irradiation device includes a laser light irradiation device 4 that emits the laser light described in the fourth embodiment, and a transmission illumination device 90 that emits transmission illumination light. The half mirror 91 that combines the laser light emitted from the laser light irradiation device 4 and the transmitted illumination light emitted from the transmission illumination device 90, the sample stage 93 on which the observation sample 92 is placed, and illumination with the transmitted light An objective lens 94 for observing the observed specimen 92, an epi-illuminator 96 for emitting epi-illumination light, a half mirror 95 for separating the epi-illumination light and the light emitted from the observation specimen 92, and And a switching mirror 97 for switching the light to the light receiving unit 98 side and the reflection mirror 99 side. The light receiving unit 98 is realized by a condenser lens, a CCD camera, or the like.

  As shown in FIG. 18, in the inverted microscope apparatus, since the distance between the transmission illumination device 90 and the observation sample 92 is generally long, the half mirror 91 can be provided between the transmission illumination device 90 and the observation sample 92. As a result, it is possible to irradiate the observation sample 92 with the transmitted illumination light emitted from the transmitted illumination device 90 and the laser light emitted from the laser beam irradiation device 4.

  This inverted microscope apparatus 9 with a laser beam irradiation device can perform surface irradiation and point irradiation even when a specimen is observed with the epi-illumination device 96.

  In the tenth embodiment, both the transmission illumination device 90 and the epi-illumination device 96 are provided, but either one of the illumination devices may be provided. In the eighth embodiment, the laser light irradiation device 4 is provided. However, the laser light irradiation device 5 shown in the fourth embodiment may be provided instead of the laser light irradiation device 4.

(Embodiment 11)
Next, a stereoscopic microscope apparatus with a laser beam irradiation apparatus according to an eleventh embodiment of the present invention will be described. FIG. 19 is a block diagram showing a schematic configuration of a stereoscopic microscope apparatus 100 with a laser beam irradiation apparatus according to an eleventh embodiment of the present invention. As shown in FIG. 19, the stereoscopic microscope apparatus 100 with a laser beam irradiation apparatus includes an irradiation unit 10 described in the first embodiment, a CCD camera 101, a binocular eye unit 102, an epi-illumination unit 103, an objective The lens unit 104, the wave plate unit 105, the transmission illumination device 107, and the detachable half mirror 108 are included.

  The binocular eyepiece 102 includes a reflection mirror 102a and imaging lenses 102bL and 102bR. The epi-illumination unit 103 includes analyzers 103aL and 103aR, deflection beam splitters (PBS) 103bL and 103bR, and polarizers 103cL and 103cR. And condensing lenses 103dL and 103dR and light guiding fibers 103eL and 103eR. The objective lens unit 104 has zoom lenses 104aL and 104aR and an objective lens 104b, and the wave plate unit 105 has a quarter wave plate 105a.

  The CCD camera 101 is attached to the right eyepiece optical path, and the position of the CCD camera 101 and the position of the image point 109 of the observation sample 106 are formed by the binocular portion 102, the epi-illumination unit 103, and the objective lens unit 104. The optical systems are in a conjugate relationship with each other.

  The half mirror 108 is inserted on the left eyepiece optical path, and the position of the beam stop 10b of the irradiation unit 10 and the position of the image point 109 of the observation sample 106 are relative to the optical system formed by the irradiation lens 11 and the half mirror 108. Therefore, they are in a conjugate positional relationship.

  First, depending on the sample to be observed, the transmission illumination device 107 or the epi-illumination device 103 is turned on and the sample is focused. The irradiation unit 10 emits point irradiation stimulation light and surface irradiation spot laser light. During transmitted illumination, the light emitted from the transmitted illumination device 107 passes through the observation sample 106 and enters the CCD camera 101 via the wave plate unit 105, the objective lens unit 104, the epi-illumination unit 103, and the binocular part 102 in order. To do. The entire image of the observation sample 106 is projected on the CCD camera 101 by the transmitted light of the transmission illumination device 107, and the entire observation sample 106 is positioned based on this image.

  Then, after irradiating guide light from the irradiation device 10 and forming a surface irradiation image on the sample, the XY movable mechanism 41 is moved to position the site to be stimulated. The point irradiation stimulation light is concentric with respect to the center of the surface irradiation image, and if the stimulation light is introduced into the fiber of the irradiation device with the light emission surface of the fiber end positioned at the aperture stop 10b, the specified point can be accurately obtained. Stimulation can be performed.

  In the eleventh embodiment, the irradiation unit 10 for irradiating laser light can be simply attached to an existing stereomicroscope device, and a desired site can be stimulated with respect to the observation sample 106.

  Further, instead of the irradiation unit 10, the irradiation unit 25 described in the second embodiment may be attached.

(Embodiment 12)
Next, the twelfth embodiment will be described. In the eleventh embodiment, the point irradiation and the surface irradiation in the irradiation units 10 and 25 are individually changed, the laser beam is emitted, the XY movable mechanism 41 is moved, and the like in the twelfth embodiment. The operation is performed in conjunction with the control of the control unit.

  FIG. 20 is a block diagram showing a schematic configuration of a stereoscopic microscope apparatus with a laser beam irradiation apparatus according to the twelfth embodiment of the present invention. As shown in FIG. 20, the stereomicroscope device with a laser beam irradiation device 120 includes an instruction unit 121, a control unit 131, drive units 131b to 131e, and shutters 11b and 103f. A setting table 131a is stored. Other configurations are the same as those of the stereomicroscope device 100 with a laser beam irradiation apparatus shown in the tenth embodiment, and the same components are denoted by the same reference numerals.

  The instruction unit 121 outputs an instruction signal including illumination information and stimulation information for the observation sample 106 to the control unit 131, and the control unit 131 sets parameters corresponding to the contents related to illumination and stimulation included in the instruction signal to the setting table 131a. And control the output of this parameter to the drive units 131b to 131e.

  Under the control of the control unit 131, the drive unit 131b opens and closes the shutter 11b to control the emission of the laser light. The drive unit 131 c drives the XY movable mechanism 41 under the control of the control unit 131 to vary the position of the image point 109 of the laser light emitted from the irradiation unit 10. The drive unit 131d drives the slider of the irradiation unit 10 under the control of the control unit 131 to variably form a point light source and a surface light source. The drive unit 131e drives an insertion / removal mechanism (not shown) of the half mirror 108 under the control of the control unit 131 to irradiate the observation sample 106 with the laser light emitted from the irradiation unit 10. To control.

  In this way, when the control unit 131 controls the shutter 11b, the XY movable mechanism 41, the irradiation unit 10, and the half mirror 108 via the drive units 131b to 131e, the desired unit indicated by the instruction unit 121 is designated. By illuminating the observation sample 106 with the illumination, positioning can be performed and stimulation can be given to a desired site.

  In the twelfth embodiment, an instruction unit 121, a control unit 131, driving units 131b to 131e, and shutters 11b and 103f are provided, and a desired part of the observation sample 106 is illuminated only by the instruction unit 121. Then, positioning can be performed to give a stimulus to a desired site.

  In the twelfth embodiment, the control unit 131 controls the irradiation unit 10, the XY movable mechanism 41, the shutters 11b and 103f, and the half mirror 108. However, the control unit 131 can move the sample stage on which the observation sample 106 is placed, or You may make it control lighting of the permeation | transmission illumination apparatus 107, etc. FIG.

  Further, in the twelfth embodiment, the control unit 131 organically functions each variable function of the stereomicroscope device with a laser beam irradiation device 120 to give a desired illumination and a desired stimulus. The control unit corresponding to 131 and the drive unit corresponding to the drive units 131b to 131e are provided in the laser beam irradiation devices 1 to 7, the microscope device 8 with the laser beam irradiation device, and the inverted microscope device 9 with the laser beam irradiation device. Also good.

  Further, the ND filter 19b and the like described in the eighth embodiment are arranged at the front end of the laser beam emission side of the irradiation unit 10 attached to the stereo microscope device 120 with the laser beam irradiation device, and the control unit 131 controls this. Thus, the amount of laser light emitted from the irradiation unit 10 may be controlled.

  Alternatively, the setting table 131a stored in the control unit 131 may be rewritable and replaced with a new parameter as needed.

It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 1 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 1 of this invention. It is a schematic diagram which shows light quantity distribution in the light emission point of the laser beam irradiation apparatus concerning Embodiment 1 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 2 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 2 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 3 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 4 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 4 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 5 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 6 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 7 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 7 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning the modification of Embodiment 7 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning the modification of Embodiment 7 of this invention. It is a block diagram of the laser beam irradiation apparatus concerning Embodiment 8 of this invention. It is a block diagram of the microscope apparatus with a laser beam irradiation apparatus concerning Embodiment 9 of this invention. It is a block diagram of the microscope apparatus with a laser beam irradiation apparatus concerning the modification of Embodiment 9 of this invention. It is a block diagram of the inverted microscope apparatus with a laser beam irradiation apparatus concerning Embodiment 10 of this invention. It is a block diagram of the stereomicroscope apparatus with a laser beam irradiation apparatus concerning Embodiment 11 of this invention. It is a block diagram which shows schematic structure of the stereomicroscope apparatus with a laser beam irradiation apparatus concerning Embodiment 12 of this invention.

Explanation of symbols

1, 1A, 2, 2A, 3, 4, 5, 6, 7 Laser light irradiation device 8, 8A Microscope device with laser light irradiation device 9 Inverted microscope device with laser light irradiation device 10, 25 Irradiation unit 10A Revolver 10a, 25a Ejection unit 10b, 10c, 25b Luminous aperture 11, 26 Irradiation lens 11b 103f Shutter 12, 27 Optical fiber 13 Slider 14 Fixing screw 15 Stop plate 16, 36, 36a, 36b, 40, 66, 78, 86, 109 Image point 17 Mirror frame 19 Lens 19a Band pass filter 19b ND filter 19c Shutter 19d, 31, 83, 103dL, 103dR, 109 Condensing lens 19e Absorption filter 20 Press ring 21, 28 Light emitting point 22, 37 Object point 23, 38, 64, 76 Condensing point 29 Collimating lens 32 Insertion / removal unit 34,63,75,87 Relay lens 35,3 a, 35b Projection lens 35A Projection lens unit 39, 82A,, 84 Dichroic mirror 88, 81, 91, 95, 108 Half mirror 41 XY movable mechanism 65, 77, 94 Condenser lens 80, 101 CCD camera 81A Concave lens 82 Observation Lens tube 84A Mirror cube 85, 94, 104b Objective lens 89 Mercury lamp 90, 107 Transmission illumination device 92, 106 Observation sample 93 Sample stage 96 Epi-illumination device 98 Light receiving unit 99, 102a, Reflection mirror 100, 120 With laser light irradiation device Stereomicroscope device 102 Binocular portion 102bL, 102bR Imaging lens 103 Epi-illumination unit 103aL, 103aR Analyzer 103bL, 103bR Polarizing beam splitter 103cL, 103cR Polarizer 103eL, 103eR Driver 104 objective lens unit 104aL, 104aR zoom lens 105 wave plate unit 105a 1/4 wavelength plate 121 instructing unit 131 control unit 131a setting table 131b, 131c, 131d, 131e driver

Claims (29)

An illumination lens system in which an IO distance, which is an interval between an object point and an image point, is set to a fixed value;
A light guiding means for guiding laser light from the light source as a point light source;
A beam stop disposed at an object point position of the irradiation lens system;
Switchable between a first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop and a second state in which the light from the point light source is irradiated so as to satisfy the diameter of the light beam stop. With an injection unit
The point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the light beam stop is projected at the image point position in the second state. Laser light irradiation device. The injection unit is
A first state in which the point light source is positioned at the object point position of the irradiation lens system; and a second state in which the point light source is positioned at a position where a divergent light beam from the point light source satisfies the light beam aperture diameter; The laser beam irradiation apparatus according to claim 1, further comprising switching means for switching between the two. The injection unit is
A collimating lens for converting a divergent light beam from the point light source into a parallel light beam having a diameter larger than the effective diameter of the light beam stop;
A condensing lens that condenses the parallel luminous flux from the collimating lens at the object point position of the irradiation lens system;
Switching means for switching between the first state in which the condenser lens is inserted into the optical axis and the second state in which the condenser lens is removed from the optical axis;
The laser light irradiation apparatus according to claim 1, comprising: An optical path branching unit is provided between the irradiation lens system and the light beam stop,
A second light beam stop disposed on the branched optical path at the object point position of the irradiation lens system;
A second exit unit for the second beam stop;
The laser beam irradiation apparatus according to claim 1, further comprising:   4. An irradiation position adjusting means for moving the light beam stop and the emission unit integrally with each other within a plane orthogonal to the optical axis with respect to the irradiation lens system. The laser beam irradiation apparatus described in 1.   The laser beam irradiation apparatus according to claim 1, further comprising a diaphragm variable unit that varies a shape or a size of the light beam diaphragm.   The laser according to claim 1, wherein the irradiation lens system is an infinity correction optical system, and the lens on the image point side can be replaced with a lens having a different focal length. Light irradiation device.   The irradiation lens system has the same configuration as the Koehler illumination optical system of a microscope, and the position of the light beam stop is a position of the field stop of the Koehler illumination, and the position of the point light source in the second state is a position that satisfies the light beam stop diameter. A laser beam irradiation apparatus characterized by that.   The laser light irradiation apparatus according to claim 1, wherein the light source includes a shutter, a wavelength selection unit, and a light amount adjustment unit.   The laser light irradiation according to claim 1, wherein the light guide unit includes an optical fiber that guides laser light, and uses an emission end of the optical fiber as the point light source. apparatus.   The laser light irradiation apparatus according to claim 10, wherein the optical fiber is a single mode fiber. The laser light source oscillates two types of laser beams having different wavelengths,
The light guide means includes two optical fibers that individually transmit the two types of laser beams,
The laser light irradiation apparatus according to claim 4, wherein the two optical fibers individually guide laser beams to the two emission units. In an upright or inverted microscope apparatus equipped with an objective lens, a specimen mounting stage, and a Kohler illumination device,
An illumination lens system in which an IO distance, which is an interval between an object point and an image point, is set to a fixed value;
A light guiding means for guiding laser light from the light source as a point light source;
A beam stop disposed at an object point position of the irradiation lens system;
A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop, and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy an effective diameter thereof. With a switchable injection unit,
Incorporating a laser light irradiation device in which the point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the beam stop is projected at the image point position in the second state. It is a microscope device with a laser beam irradiation device,
An optical path branching unit for branching the optical path of the Koehler illumination device is provided,
Placing the light beam stop and the exit unit on the optical path branched by the optical path branching means;
The position of the light beam stop is a position conjugate with the field stop of the Koehler illumination device, and the position of the point light source in the second state is a position satisfying the diameter of the light beam stop, and as a part of the irradiation lens system A microscope apparatus with a laser beam irradiation apparatus, characterized by using an objective lens and a light projection lens. The injection unit is
The first state where the point light source is positioned at the object point position of the irradiation lens system, and the point light source is positioned at a position where the divergent light beam from the point light source sufficiently satisfies the effective diameter of the light beam stop. 14. The microscope apparatus with a laser beam irradiation apparatus according to claim 13, further comprising switching means for switching between two states. The injection unit is
A collimating lens for converting a divergent light beam from the point light source into a parallel light beam having a diameter larger than the effective diameter of the light beam stop;
A condensing lens that condenses the parallel luminous flux from the collimating lens at the object point position of the irradiation lens system;
Switching means for switching between the first state in which the condenser lens is inserted into the optical axis and the second state in which the condenser lens is removed from the optical axis;
The microscope apparatus with a laser beam irradiation apparatus according to claim 13.   16. The microscope apparatus with a laser beam irradiation apparatus according to claim 13, wherein an optical path extension lens is provided on the optical path branched by the optical path branching unit. In an upright or inverted microscope apparatus equipped with an objective lens, a specimen mounting stage, and an observation optical path,
An illumination lens system in which an IO distance, which is an interval between an object point and an image point, is set to a fixed value;
A light guiding means for guiding laser light from the light source as a point light source;
A beam stop disposed at an object point position of the irradiation lens system;
A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop, and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy an effective diameter thereof. With a switchable injection unit,
Incorporating a laser light irradiation device in which the point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the beam stop is projected at the image point position in the second state. It is a microscope device with a laser beam irradiation device,
An optical path branching unit for branching the observation optical path is provided,
Placing the light beam stop and the exit unit on the optical path branched by the optical path branching means;
A microscope with a laser beam irradiation apparatus, wherein the position of the light beam stop is a position conjugate with the imaging position of the observation optical path, and the objective lens and the imaging lens are used as a part of the irradiation lens system. apparatus. The injection unit is
The first state where the point light source is positioned at the object point position of the irradiation lens system, and the point light source is positioned at a position where the divergent light beam from the point light source sufficiently satisfies the effective diameter of the light beam stop. 18. The microscope apparatus with a laser beam irradiation apparatus according to claim 17, further comprising switching means for switching between two states. The injection unit is
A collimating lens for converting a divergent light beam from the point light source into a parallel light beam having a diameter larger than the effective diameter of the light beam stop;
A condensing lens that condenses the parallel luminous flux from the collimating lens at the object point position of the irradiation lens system;
Switching means for switching between the first state in which the condenser lens is inserted into the optical axis and the second state in which the condenser lens is removed from the optical axis;
The microscope apparatus with a laser beam irradiation apparatus according to claim 17, comprising: In an inverted microscope device equipped with an objective lens, a specimen stage, and a transmission illumination device,
An illumination lens system in which an IO distance, which is an interval between an object point and an image point, is set to a fixed value;
A light guiding means for guiding laser light from the light source as a point light source;
A beam stop disposed at an object point position of the irradiation lens system;
A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop; and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy the effective diameter thereof. A switchable injection unit,
Incorporating a laser light irradiation apparatus in which the point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the light beam stop is projected at the image point position in the second state. It is a microscope device with a laser beam irradiation device,
An optical path branching unit is provided between the sample stage and the condenser lens of the transmission illumination device,
On the optical path branched by the optical path branching means, the irradiation lens system, the light beam stop, and the emission unit are arranged,
A microscope apparatus with a laser beam irradiation apparatus, wherein an image point position of the irradiation lens system coincides with a focus position of the objective lens. The injection unit is
The first state where the point light source is positioned at the object point position of the irradiation lens system, and the point light source is positioned at a position where the divergent light beam from the point light source sufficiently satisfies the effective diameter of the light beam stop. 21. The microscope apparatus with a laser beam irradiation apparatus according to claim 20, further comprising switching means for switching between two states. The injection unit is
A collimating lens for converting a divergent light beam from the point light source into a parallel light beam having a diameter larger than the effective diameter of the light beam stop;
A condensing lens that condenses the parallel luminous flux from the collimating lens at the object point position of the irradiation lens system;
Switching means for switching between the first state in which the condenser lens is inserted into the optical axis and the second state in which the condenser lens is removed from the optical axis;
The microscope apparatus with a laser beam irradiation apparatus according to claim 20. Imaging means for imaging an object;
Display means for displaying the imaged observation image;
Position designation means for designating an arbitrary point of the observation image on the display means;
An irradiation position adjusting means for integrally moving the light beam stop and the emission unit in the XY directions orthogonal to the optical axis with respect to the irradiation lens system;
Wavelength switching means for switching the wavelength of the laser light to the stimulation light to the specimen and the guide light to which the specimen does not react;
Irradiation light setting means for setting the type of laser light applied to the specimen as a combination of one of stimulation light or guide light and one of the point light source image or the image of the light beam stop,
The wavelength switching unit and the switching unit of the emission unit are controlled based on the type of irradiation light set by the wavelength switching unit, and the irradiation position adjusting unit is controlled based on the position specified by the position specifying unit. Control means,
The microscope apparatus with a laser beam irradiation apparatus according to any one of claims 13 to 22, wherein a laser beam of a type set at a specified position is irradiated. Further comprising a light control means for adjusting the amount of the laser light,
The irradiation light setting means can set the amount of laser light to be irradiated,
24. The microscope apparatus with a laser light irradiation apparatus according to claim 23, wherein the control means controls the light control means based on the set amount of laser light. An irradiation lens system in which an I-O distance, which is a distance between an object point and an image point, is set to a fixed value in a stereomicroscope device provided with a TV optical path on one of the two observation optical axes;
A light guiding means for guiding laser light from the light source as a point light source;
A beam stop disposed at an object point position of the irradiation lens system;
A first state in which the point light source or a projected image of the point light source is formed at the position of the light beam stop, and a second state in which the light beam stop is irradiated with light from the point light source so as to satisfy an effective diameter thereof. With a switchable injection unit,
Incorporating a laser light irradiation apparatus in which the point light source is projected at the image point position of the irradiation lens system in the first state, and the shape of the light beam stop is projected at the image point position in the second state. It is a stereo microscope device with a laser beam irradiation device,
An optical path branching means is provided between the objective lens and the sample and on the optical path on the side where the TV optical path is not provided,
On the optical path branched by the optical path branching means, the irradiation lens system, the light beam stop, and the emission unit are arranged,
A microscope apparatus with a laser beam irradiation apparatus, wherein an image point position of the irradiation lens system coincides with a focus position of an objective lens. The injection unit is
The first state where the point light source is positioned at the object point position of the irradiation lens system, and the point light source is positioned at a position where the divergent light beam from the point light source sufficiently satisfies the effective diameter of the light beam stop. 26. The microscope apparatus with a laser beam irradiation apparatus according to claim 25, further comprising switching means for switching between two states. The injection unit is
A collimating lens for converting a divergent light beam from the point light source into a parallel light beam having a diameter larger than the effective diameter of the light beam stop;
A condensing lens that condenses the parallel luminous flux from the collimating lens at the object point position of the irradiation lens system;
Switching means for switching between the first state in which the condenser lens is inserted into the optical axis and the second state in which the condenser lens is removed from the optical axis;
The microscope apparatus with a laser beam irradiation apparatus according to claim 25, comprising: Imaging means for imaging an object;
Display means for displaying the imaged observation image;
Position designation means for designating an arbitrary point of the observation image on the display means;
An irradiation position adjusting means for integrally moving the beam stop and the emission unit in an XY direction perpendicular to the optical axis with respect to the irradiation lens system;
Irradiation light setting means for setting the wavelength of the laser light as a combination of one of the stimulation light to the sample and the guide light that the sample does not react with, one of the point light source image or the image of the light beam stop,
The wavelength switching unit and the switching unit of the emission unit are controlled based on the type of irradiation light set by the wavelength switching unit, and the irradiation position adjusting unit is controlled based on the position specified by the position specifying unit. Control means,
28. The microscope apparatus with a laser beam irradiation apparatus according to claim 25, wherein a laser beam of a type set at a designated position is irradiated. Further comprising a light control means for adjusting the amount of the laser light,
The irradiation light setting means can set the amount of laser light to be irradiated,
29. The microscope apparatus with a laser light irradiation apparatus according to claim 28, wherein the control means controls the light control means based on the set amount of laser light.

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