JP4464654B2 - Microscope equipment - Google Patents

Description

  The present invention relates to a microscope apparatus that enables a plurality of observation methods by switching optical elements.

  Conventionally, as a microscope apparatus, excitation light from a light source is condensed on a specimen by an objective lens, and fluorescence emitted from the specimen is condensed by an imaging lens through the objective lens, and by a prism disposed in a lens barrel. There is known a fluorescence microscope which is deflected to the eyepiece lens side so that a fluorescence image can be visually observed by the eyepiece lens.

  Some of these fluorescent microscopes are provided with an imaging device mounting portion for mounting an imaging device such as a CCD camera for imaging fluorescence transmitted through a prism in the lens barrel above the lens barrel.

  On the other hand, recently, a confocal scanning microscope that two-dimensionally scans a sample with light from a point light source and detects light from the sample through a pinhole or slit provided on a rotating disk has attracted attention. However, by placing a confocal device equipped with a rotating disk of such a microscope on the imaging device mounting portion provided on the above-described fluorescent microscope barrel, the function of a confocal scanning microscope is added to a normal fluorescent microscope. The thing which added it is also considered.

  However, a microscope apparatus configured in this way generally has a confocal device having a rotating disk and an imaging device arranged above the lens barrel, so that the center of gravity of the entire device becomes high and installation is difficult. It becomes stable. In addition, if the confocal device is attached to the imaging device mounting portion provided in the lens barrel of the fluorescence microscope, a pinhole must be disposed at the focal position of the imaging lens, so the relative position of the confocal device with respect to the lens barrel The relationship is also practically limited.

  For this reason, it is difficult to add other application devices between the lens barrel and the confocal device as well as the periphery of the lens barrel, and there is a problem that the expandability of the system is limited.

  The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a microscope apparatus that is excellent in expandability and that does not apply unnecessary light to a specimen.

  The invention according to claim 1 is provided on a side surface of the microscope main body having a microscope main body having an objective lens arranged close to the specimen, and an observation means for observing the optical image of the specimen via the objective lens, A light projecting tube having at least one imaging means, a wavelength selection optical element that reflects light from the light source to the specimen and transmits light from the specimen to the observation means, and light from the light source. A first optical element switching unit that selectively inserts a reflective optical element that totally reflects on the specimen into the optical path; and a light that is in the light projecting tube and reflects light from the light source to the first optical element switching unit. A second optical element that selectively inserts into the optical path a wavelength selection optical element that transmits light from the specimen to the imaging means and a reflection optical element that totally reflects light from the light source to the first optical element switching means. Element switching means; and Dimming means for blocking or dimming light from the source, and when the reflecting optical elements of the first and second optical element switching means are simultaneously inserted in the optical path, the dimming means It is characterized by shielding or dimming light from the light source.

  According to a second aspect of the present invention, in the first aspect of the present invention, the light projecting tube includes a rotating disk in which light from the light source is collected and a plurality of through holes are inserted at positions conjugate with the specimen. It is characterized by being removable.

  According to a third aspect of the present invention, in the first aspect of the invention, the light projecting tube includes a laser light source, a two-dimensional scanning unit that two-dimensionally scans the specimen with laser light from the laser light source, and the specimen. It further has confocal observation means for taking in the light.

According to a fourth aspect of the present invention, in the first or third aspect of the present invention, the first and second optical element switching means detect that each of the reflective optical elements is inserted in an optical path. First and second position detecting means, and when the first and second position detecting means simultaneously detect the reflective optical element inserted into the optical path, the light control means blocks light from the light source. It is also characterized by dimming.

  According to a fifth aspect of the invention, in the first aspect of the invention, in place of the dimming means, the second optical element switching means is provided with an ultraviolet cut filter inserted into the optical path together with the reflective optical element. It is characterized by.

  According to a sixth aspect of the invention, in the first aspect of the invention, the first and second optical elements when the reflective optical elements of the first and second optical element switching means are simultaneously inserted into an optical path. One of the switching means is forcibly retracted from the optical path.

  A seventh aspect of the invention is characterized in that, in the first aspect of the invention, the light control means comprises a shutter or an ND filter.

  According to the present invention, the light projecting tube provided on the side surface of the microscope main body is provided with the imaging means, the rotating disk, and the like, so that the rotating disk and the imaging device are disposed above the lens barrel of the conventional microscope main body. The center of gravity of the entire apparatus can be lowered as compared with the arrangement in which the microscopes are stacked, and the microscope apparatus can be stably installed. In addition, since the projection tube can be attached to the microscope body without being restricted by the relative positional relationship with respect to the lens barrel as in the past, it is easy to add other application devices. The extensibility of can be improved.

  Further, according to the present invention, the first optical element switching means provided in the microscope body and the second optical element switching means provided in the light projection tube are separately switched, and the wavelength selection optical element and the reflection are switched. By selecting the combination of the optical elements, it is possible to pick up the fluorescence observation image by the image pickup means and the visual observation of the fluorescence observation image. However, the respective reflections can be made by the switching operation of the first and second optical element switching means. When the optical element is inserted into the optical path at the same time, light from the light source is blocked or dimmed by the dimming means so that unnecessary light is not applied to the specimen 3 as much as possible. It is possible to reliably avoid the inconvenience that the fluorescence of the specimen is faded by the light of the wavelength.

  Further, according to the present invention, in place of the light control means, the second optical element switching means is provided with an ultraviolet cut filter inserted into the optical path together with the reflective optical element, so that the wavelength in the ultraviolet region that most affects the fading of fluorescence. Therefore, fading of the fluorescence of the specimen can be reduced.

  Furthermore, according to the present invention, when the respective reflecting optical elements are simultaneously inserted into the optical path by the switching operation of the first and second optical element switching means, either one of the reflecting optical elements is forcibly inserted into the optical path. Therefore, the light from the light source can be effectively shielded, and unnecessary fading of the specimen can be avoided.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

(First embodiment)
FIG. 1 shows a schematic configuration of an example in which the present invention is applied to an inverted microscope apparatus. In the figure, reference numeral 1 denotes a microscope body, and a stage 2 is disposed on the microscope body 1. A specimen 3 is placed on the stage 2.

  The microscope main body 1 is provided with an illumination column 4 facing the stage 2 above. The front end of the illumination column 4 is bent at a substantially right angle, and the front end is disposed in a horizontal direction with respect to the surface of the stage 2.

  A light source 5 for transmitted illumination is provided at the tip of the illumination column 4. As the light source 5, a halogen lamp, a mercury lamp, or the like is used.

  In addition, a capacitor 6 is provided at the tip of the illumination column 4. The capacitor 6 is disposed on the illumination light path of the light source 5 and condenses the illumination light from the light source 5 at the position of the sample 3.

  Below the stage 2, an objective lens 7 is disposed close to the sample 3. A plurality of objective lenses 7 having different magnifications (only one is shown in the drawing) are held by a revolver (not shown), and are selectively positioned on the observation optical path a by rotating the revolver. It has become.

The objective lens 7 is moved up and down along the observation optical path a together with the revolver by operating a focusing handle (not shown), and the relative distance from the stage 2 is changed so that the specimen 3 can be focused. A reflection mirror 8 is arranged on the observation light path a below the display 7. The reflection mirror 8 reflects the light image that has passed through the specimen 3 and is magnified by the objective lens 7 in an obliquely upward direction (an angle of 45 ° with respect to the horizontal). Then, the light image reflected by the reflecting mirror 8 is relayed by the relay optical system 9, enters the eyepiece 11 as an observation means attached to the observation barrel 10, and is visually observed by the observer. ing. In this case, the observation lens barrel 10 to which the eyepiece 11 is attached is disposed in front of the microscope body 1.

  A light projecting tube 12 is provided on the side surface of the microscope main body 1, here on the back surface opposite to the front surface of the microscope main body 1 on which the observation barrel 10 is provided. In this case, the light projecting tube 12 is provided on the back surface of the microscope body 1 so as to protrude in the horizontal direction.

  A light source 13 is provided in the light projecting tube 12. The light source 13 is a xenon or mercury lamp. On the optical path of the illumination light emitted from the light source 13, a collector lens 14 and a cube turret 15 as second optical element switching means are arranged. The cube turret 15 can hold a plurality of types of cube units, and can be manually or automatically rotated. In the illustrated example, it is assumed that two types of cube units 16 and 17 are mounted, and the cube unit 16 is positioned on the optical path of the illumination light from the collector lens 14.

  In this case, the cube unit 16 is provided with a dichroic mirror 16a as a wavelength selection optical element. The dichroic mirror 16a is a kind of interference filter, has a characteristic of reflecting a short wavelength and transmitting a long wavelength, reflects a wavelength of excitation light for exciting the sample 3, and emits fluorescence emitted from the sample 3. The wavelength is transmitted. The cube unit 17 is provided with a total reflection mirror 17a as a reflection optical element. The total reflection mirror 17 a reflects all of the illumination light from the light source 13.

  The cube turret 15 is provided with a magnet 18 a and a magnetic sensor 18 b as the first position detecting means 18. The magnet 18a and the magnetic sensor 18b detect that the total reflection mirror 17a of the cube unit 17 is inserted in the optical path.

  A rotating disk 19 and an imaging lens 20 are disposed in the reflected light path of the dichroic mirror 16a (positioned on the light path from the collector lens 14) of the cube unit 16. The rotating disk 19 is disposed on a point I that is a position where the excitation light collected by the collector lens 14 is conjugated with the sample 3 and is rotatable about a rotating shaft 19a. The rotating disk 19 is formed with a plurality of through holes (not shown) such as pin holes or slits, and excitation light that has passed through these through holes enters the imaging lens 20. Further, the rotating disk 19 can be inserted into and removed from the optical path. In the illustrated example, the rotating disk 19 is in a state of being retracted from the optical path. The imaging lens 20 is image-side telecentric so that the light passing therethrough becomes parallel light.

  An imaging lens 21 that constitutes an imaging optical system is disposed in the transmitted light path of the dichroic mirror 16a of the cube unit 16 (positioned on the optical path from the collector lens 14). At the imaging position of the imaging lens 21, a CCD camera 22 as an imaging means is disposed. The fluorescence from the specimen 3 that has passed through the imaging lens 21 is guided to the CCD camera 22 and imaged on the imaging surface 22a.

  On the optical path from the imaging lens 20, a cube turret 23 is arranged as a first optical element switching means at a point where it intersects with the observation optical path a inside the microscope body 1. The cube turret 23 can hold a plurality of types of cube units, and can be manually or automatically rotated. In the illustrated example, it is assumed that two types of cube units 24 and 25 are mounted, and the cube unit 24 is positioned at the intersection of the optical path from the imaging lens 20 and the observation optical path a. In this case, the cube unit 24 is provided with a total reflection mirror 24a as a reflection optical element. The total reflection mirror 24a reflects all the light from the imaging lens 20 to the objective lens 7 side. The cube unit 25 is provided with a dichroic mirror 25a as a wavelength selection optical element. The dichroic mirror 25a has a characteristic of reflecting a short wavelength and transmitting a long wavelength, reflects an excitation light wavelength for exciting the sample 3, transmits a fluorescence wavelength emitted by the sample 3, The light is guided to the eyepiece lens 11 through the reflection mirror 8 described above.

  The cube turret 23 is provided with a magnet 26 a and a magnetic sensor 26 b as the second position detecting means 26. The magnet 26a and the magnetic sensor 26b detect that the total reflection mirror 24a of the cube unit 24 is inserted into the optical path.

  A shutter 27 as a light control means is provided on the front surface of the light source 13 in the light projecting tube 12. The shutter 27 can be inserted into and removed from the optical path of the illumination light from the light source 13 and can shield the illumination light as necessary. A drive control unit 28 is connected to the shutter 27. In this drive control unit 28, the magnetic sensor 18b of the first position detecting means 18 and the magnetic sensor 26b of the second position detecting means 26 simultaneously detect the total reflection mirrors 17a and 24a inserted in the optical path, respectively. When the output is generated, the shutter 27 is inserted into the optical path to block the illumination light from the light source 13. Of course, when the detection output of either the magnetic sensor 18b or the magnetic sensor 26b is stopped, the drive control unit 28 again avoids the shutter 27 from the optical path.

  When the first and second position detectors 18 and 26 simultaneously detect the total reflection mirrors 17a and 24a inserted in the optical path, an alarm is issued and the shutter 27 is manually inserted in the optical path. You may make it prompt.

  Next, the operation of the embodiment configured as described above will be described.

  First, a case where a fluorescence observation image is captured using the CCD camera 22 will be described.

  In this case, as shown in FIG. 1, the cube turret 15 inserts the dichroic mirror 16a of the cube unit 16 into the optical path, and the cube turret 23 inserts the total reflection mirror 24a of the cube unit 24 into the optical path.

  In this state, the light generated from the light source 13 is incident on the dichroic mirror 16a via the collector lens 14. The dichroic mirror 16a reflects excitation light for exciting the fluorescent reagent of the specimen 3. The excitation light reflected by the dichroic mirror 16 a is guided to the imaging lens 20, converted into parallel light, folded back by the total reflection mirror 24 a, and condensed and irradiated onto the sample 3 through the objective lens 7.

  When fluorescence is emitted from the specimen 3, the fluorescence wavelength at this time is shifted to a longer wavelength side than the wavelength of the excitation light by Stokes shift, and is reflected again by the total reflection mirror 24a via the objective lens 7. The light enters the image lens 20. Then, a position conjugate with the sample 3 is formed at the indicated point I, passes through the dichroic mirror 16a, passes through the imaging lens 21, and forms an image on the imaging surface 22a of the CCD camera 22, and a fluorescence observation image is captured.

  In this case, if the rotating disk 19 is inserted at the point I conjugate with the sample 3 and rotated around the rotating shaft 19a, confocal observation by the CCD camera 22 becomes possible, and the rotating disk 19 is removed from the optical path. By retracting, normal fluorescence observation by the CCD camera 22 can be performed. Further, if the cube unit 16 is replaced with another cube unit (not shown) and a dichroic mirror having different characteristics is used, other fluorescent reagents of the specimen 3 can be dealt with.

  Next, the case where the fluorescence observation image is visually observed using the eyepiece 11 will be described.

  In this case, the total reflection mirror 17a of the cube unit 17 is inserted into the optical path by the cube turret 15 as shown in FIG. Thus, the dichroic mirror 25a of the cube unit 25 is inserted into the optical path.

  In this state, the light generated from the light source 13 is incident on the total reflection mirror 17a via the collector lens 14, is all turned back, and is incident on the dichroic mirror 25a via the imaging lens 20. The dichroic mirror 25a reflects excitation light for exciting the fluorescent reagent of the specimen 3. The excitation light reflected by the dichroic mirror 25a is condensed and irradiated on the specimen 3 through the objective lens 7.

  When fluorescence is emitted from the specimen 3, the fluorescence wavelength at this time is shifted to a longer wavelength side than the wavelength of the excitation light by Stokes shift, enters the dichroic mirror 25a through the objective lens 7, and enters the dichroic mirror 25a. And enters the reflection mirror 8. Then, it is folded back by the reflecting mirror 8, relayed by the relay optical system 9, and incident on the eyepiece 11 of the observation barrel 10, so that fluorescence observation can be performed by the observer's visual observation.

  In this case, if the cube unit 25 is replaced with another cube unit (not shown) and a dichroic mirror having different characteristics is used, other fluorescent reagents of the specimen 3 can be handled.

  By the way, in such a microscope apparatus, due to an operational error, the total reflection mirror of the cube unit 17 is caused by the cube turret 15 as shown in FIG. The total reflection mirror 24a of the cube unit 24 may be inserted into the optical path by the cube turret 23 in a state where 17a is inserted into the optical path. That is, the total reflection mirror 17a and the total reflection mirror 24a may be simultaneously set on the optical path. In this state, although no observation can be performed, the illumination light from the light source 13 enters the total reflection mirror 17 a through the collector lens 14, is turned back, and is totally reflected through the imaging lens 20. The light is incident on the mirror 24 a, is further turned back by the total reflection mirror 24 a, and is focused and irradiated on the sample 3 through the objective lens 7. In other words, under this situation, all of the illumination light from the light source 13 is irradiated onto the specimen 3, and there is a possibility that the fluorescence of the specimen 3 may rapidly fade due to short-wavelength light in the illumination light.

  On the other hand, in the first embodiment, when the total reflection mirror 17a of the cube unit 17 is inserted into the optical path by the cube turret 15, an output is generated from the magnetic sensor 18b of the first position detecting means 18, and When the total reflection mirror 24a of the cube unit 24 is inserted into the optical path by the cube turret 23, an output is generated from the magnetic sensor 26b of the second position detection means 26. And when these magnetic sensors 18b and 26b generate | occur | produce output simultaneously, the illumination light from the light source 13 is light-shielded by the shutter 27 being inserted in the optical path by the drive control part 28. FIG.

  Thereafter, if the output of either the magnetic sensor 18b or the magnetic sensor 26b is stopped, that is, if either one of the total reflection mirror 17a or the total reflection mirror 24a is removed from the optical path, the shutter 27 is moved by the drive control unit 28. Avoided from the light path.

  Accordingly, in this manner, the light source 13, the CCD camera 22, the cube turret 15 having the cube units 16 and 17, and the rotating disk 19 are provided on the light projecting tube 12 provided on the side surface of the microscope body 1. Compared to conventional confocal devices and imaging devices equipped with a rotating disk above the microscope barrel of the microscope body, the center of gravity of the entire device can be lowered, and the microscope device can be stabilized. Can be installed.

  Further, since the projection tube 12 can be attached to the microscope body 1 without being restricted by the relative positional relationship with respect to the lens barrel as in the prior art, it is easy to add other application devices. The expandability of the device can be improved.

  Further, the cube turret 15 provided in the light projecting tube 12 and the cube turret 23 provided in the microscope main body 1 are switched, and a fluorescence observation image is captured by the CCD camera 22 by combining the dichroic mirror 16a and the total reflection mirror 24a. In addition, it is possible to visually observe the fluorescence observation image by combining the total reflection mirror 17a and the dichroic mirror 25a. However, the total reflection mirror 17a can be switched by switching the cube turrets 15 and 23 due to an operational error. , 24a are selected at the same time, the shutter 27 is moved along the optical path in accordance with the outputs of the first position detecting means 18 and the second position detecting means 26 for detecting the total reflection mirrors 17a, 24a. Since the illumination light from the light source 13 is shielded, the light source Fluorescence of the sample 3 by short-wavelength light included in the illumination light 3 can be reliably avoid a disadvantage that would fade.

  In the above-described embodiment, the illumination light is shielded by using the shutter 27. However, the illumination light from the light source 13 may be shielded by inserting and removing the rotary disk 19 in the optical path, or an ND filter. Substituting with dimming means such as Also, when the cube turrets 15 and 23 are of the electric drive system, when one is set as a total reflection mirror and the other is set as a total reflection mirror, (a) operation Do not do. (B) After the total reflection mirror previously entered in the optical path is retracted from the optical path, an operation is performed in which the subsequent total reflection mirror is inserted into the optical path. (C) After the shutter 27 is inserted into the optical path, the subsequent total reflection mirror is inserted into the optical path. By performing the control as described above, the illumination light can be effectively blocked, and unnecessary fading of the specimen 3 can be avoided.

(Modification)
Next, a modification of the first embodiment will be described with reference to FIG. In FIG. 4, the same parts as those in FIG.

  In this case, the cube unit 17 attached to the cube turret 15 includes an ultraviolet cut filter 17b as an alternative to the shutter 27 in addition to the total reflection mirror 17a. The ultraviolet cut filter 17b has such a characteristic as to cut the wavelength in the ultraviolet region that most affects the fading of fluorescence contained in the illumination light of the light source 13.

  Others are the same as FIG.

  In such a configuration, when the fluorescence observation image is visually observed, the purple ultraviolet cut filter 17b is inserted into the optical path together with the total reflection mirror 17a of the cube unit 17 by the cube turret 15, and the cube unit 25 is inserted by the cube turret 23. The dichroic mirror 25a is inserted into the optical path.

  In this state, the light emitted from the light source 13 enters the ultraviolet cut filter 17b through the collector lens 14, and the wavelength in the ultraviolet region included in the illumination light is cut. Then, it is folded back by the total reflection mirror 17a and enters the dichroic mirror 25a through the imaging lens 20. The dichroic mirror 25a reflects excitation light for exciting the fluorescent reagent of the specimen 3. The excitation light reflected by the dichroic mirror 25a is condensed and irradiated on the specimen 3 through the objective lens 7. Further, the fluorescence emitted from the specimen 3 passes through the dichroic mirror 25a, is folded back by the reflection mirror 8, enters the eyepiece lens 11 of the observation barrel 10 via the relay optical system 9, and is visually observed by the observer. Observation becomes possible.

  Even in this case, even if the total reflection mirror 24a of the cube unit 24 is accidentally inserted into the optical path by the cube turret 23, the ultraviolet cut filter 17b is inserted into the optical path, and the ultraviolet ray that has the greatest influence on the fading of fluorescence. Since the wavelength of the region is cut, the fading of the fluorescence of the sample 3 can be reduced. Further, since the shutter 27 can be omitted, the configuration can be greatly simplified considering that the first position detecting means 18 and the second position detecting means 26 can be omitted.

  On the other hand, when observing ultraviolet excitation by visual observation, if the ultraviolet cut filter 17b is inserted in the optical path, it is incident from the light source 13 through a filter (filter for selecting the wavelength in the ultraviolet region) or the like (not shown). Excitation ultraviolet rays are cut off. Therefore, in this case, the dichroic mirror 16a of the cube unit 16 is inserted into the optical path by the cube turret 15 as shown in FIG. The dichroic mirror 16a is used for exciting ultraviolet light that reflects ultraviolet light and transmits light having a longer wavelength. Similarly, the dichroic mirror 25a of the cube unit 25 needs to be used for exciting ultraviolet rays. In this way, excitation by ultraviolet rays is possible and visual observation is also possible.

(Second embodiment)
Next, a second embodiment of the present invention will be described.

  FIG. 6 shows a schematic configuration of an example in which the present invention is applied to a laser scanning confocal microscope, and the same parts as those in FIG. 1 are denoted by the same reference numerals.

  In this case, the light projecting tube 12 is provided with a laser light source 31. A mirror 32 is disposed on the optical path of the laser light emitted from the laser light source 31. In addition, a dichroic mirror 33 is disposed in the reflected light path of the mirror 32. The dichroic mirror 33 has a characteristic of transmitting the wavelength of the laser light and reflecting the wavelength region of the fluorescence emitted from the sample 3.

  On the transmitted light path of the dichroic mirror 33, a two-dimensional scanning mechanism 34 as an optical scanning unit is arranged. The two-dimensional scanning mechanism 34 has two sets of galvano scanner mirrors (not shown), and these galvano scanner mirrors scan laser light in a two-dimensional direction.

  On the optical path of the laser light emitted from the two-dimensional scanning mechanism 34, the cube turret 15 is arranged at a point that intersects the optical path of the illumination light from the light source 13. In this case, the cube turret 15 has a cube unit 35 having holes in addition to the cube units 16 and 17 described above. This cube unit 35 is for making the laser beam from the two-dimensional scanning mechanism 34 directly enter the imaging lens 20 through the air holes.

  On the other hand, on the reflected light path of the dichroic mirror 33, a confocal pinhole 36 and a light detector 37 constituting a confocal observation means are arranged. Here, for example, a photomultiplier is used as the photodetector 37.

  Others are the same as FIG.

  In such a configuration, when confocal observation is performed, as shown in FIG. 6, the cube turret 15 inserts the holes of the cube unit 35 into the optical path, and the cube turret 23 uses the total reflection mirror 24a of the cube unit 24. In the light path.

  In this state, the laser light generated from the laser light source 31 is folded back by the mirror 32, transmitted through the dichroic mirror 33, scanned two-dimensionally by the two-dimensional scanning mechanism 34, and passes through the holes of the cube unit 35. The light is guided to the imaging lens 20, converted into parallel light, folded back by the total reflection mirror 24 a, and condensed and irradiated to the specimen 3 through the objective lens 7.

  Further, the fluorescence emitted from the specimen 3 passes through the objective lens 7, is turned back by the total reflection mirror 24a, is returned to linear light through the two-dimensional scanning mechanism 34, is reflected by the dichroic mirror 33, and is confocal pinhole. After passing through 36, it is detected by the photodetector 37, and imaging and analysis processing is performed.

  Next, in the case of visually observing fluorescence, as shown in FIG. 7 (the same parts as in FIG. 6 are given the same reference numerals), the cube turret 15 causes the total reflection mirror 17a of the cube unit 17 to be in the optical path. The dichroic mirror 25a of the cube unit 25 is inserted into the optical path by the cube turret 23.

  In this state, the light generated from the light source 13 is incident on the total reflection mirror 17a via the collector lens 14, is all turned back, and is incident on the dichroic mirror 25a via the imaging lens 20. The dichroic mirror 25a reflects excitation light for exciting the fluorescent reagent of the specimen 3. The excitation light reflected by the dichroic mirror 25a is condensed and irradiated on the specimen 3 through the objective lens 7.

  When fluorescence is emitted from the sample 3, the wavelength of the fluorescence at this time is shifted to the longer wavelength side than the wavelength of the excitation light by Stokes shift, enters the dichroic mirror 25a through the objective lens 7, and this dichroic mirror. The light passes through 25a and enters the reflection mirror 8. Then, it is folded back by the reflecting mirror 8, relayed by the relay optical system 9, and incident on the eyepiece 11 of the observation barrel 10, so that fluorescence observation can be performed by the observer's visual observation.

  In this case as well, due to an operational error, the cube turret 15 causes the total reflection mirror 17a of the cube unit 17 to be moved into the optical path by the cube turret 15, as shown in FIG. In some cases, the total reflection mirror 24a of the cube unit 24 may be inserted into the optical path by the cube turret 23. That is, the total reflection mirror 17a and the total reflection mirror 24a may be simultaneously set on the optical path. In this state, although no observation can be performed, the illumination light from the light source 13 enters the total reflection mirror 17 a through the collector lens 14, is turned back, and is totally reflected through the imaging lens 20. The light is incident on the mirror 24 a, is further turned back by the total reflection mirror 24 a, and is focused and irradiated on the sample 3 through the objective lens 7. That is, under this situation, all of the illumination light from the light source 13 is irradiated onto the specimen 3, and there is a possibility that the fluorescence of the specimen 3 may be faded by the short wavelength light in the illumination light.

  On the other hand, also in the second embodiment, when the total reflection mirror 17a of the cube unit 17 is inserted into the optical path by the cube turret 15, an output is generated from the magnetic sensor 18b of the first position detecting means 18, and When the total reflection mirror 24a of the cube unit 24 is inserted into the optical path by the cube turret 23, an output is generated from the magnetic sensor 26b of the second position detection means 26. When the magnetic sensors 18b and 26b generate outputs at the same time, the illumination light from the light source 13 is shielded by inserting the shutter 27 into the optical path by the drive control unit 28.

  Thereafter, if the output of either the magnetic sensor 18b or the magnetic sensor 26b is stopped, that is, if either one of the total reflection mirror 17a or the total reflection mirror 24a is removed from the optical path, the shutter 27 is moved by the drive control unit 28. Avoided from the light path.

  Therefore, even in this way, the same effect as in the first embodiment can be obtained.

  In addition, the cube turret 15 provided in the light projecting tube 12 and the cube turret 23 provided in the microscope main body 1 are switched, and the combination of the cube unit 35 having holes and the total reflection mirror 24a enables confocal observation. In addition, the fluorescent observation image can be visually observed by the combination of the total reflection mirror 17a and the dichroic mirror 25a. In this case, the total reflection mirror 17a, When an unnecessary combination that simultaneously selects 24a occurs, the illumination light from the light source 13 can be blocked by the shutter 27 in accordance with the outputs of the first position detecting means 18 and the second position detecting means 26. The inconvenience that the fluorescence of the specimen 3 is faded by the short wavelength light included in the illumination light 13 is confirmed. It can be avoided to.

  In addition, this invention is not limited to the said embodiment, In the implementation stage, it can change variously in the range which does not change the summary. For example, in the above-described embodiments, the inverted microscope apparatus has been described, but the present invention can be similarly applied to an upright microscope apparatus.

  Furthermore, the above embodiments include inventions at various stages, and various inventions can be extracted by appropriately combining a plurality of disclosed constituent elements. For example, even if some constituent requirements are deleted from all the constituent requirements shown in the embodiment, the problem described in the column of the problem to be solved by the invention can be solved, and is described in the column of the effect of the invention. If the above effect is obtained, a configuration from which this configuration requirement is deleted can be extracted as an invention.

The figure which shows schematic structure of the 1st Embodiment of this invention. The figure which shows schematic structure for demonstrating operation | movement of 1st Embodiment. The figure which shows schematic structure for demonstrating the other operation | movement of 1st Embodiment. The figure which shows schematic structure of the modification of 1st Embodiment. The figure which shows schematic structure for demonstrating the operation | movement of the modification of 1st Embodiment. The figure which shows schematic structure of the 2nd Embodiment of this invention. The figure which shows schematic structure for demonstrating operation | movement of 2nd Embodiment. The figure which shows schematic structure for demonstrating the other operation | movement of 2nd Embodiment.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Microscope main body, 2 ... Stage, 3 ... Sample, 4 Illumination support | pillar 5 ... Light source, 6 ... Condenser, 7 ... Objective lens, 8 ... Reflection mirror 9 ... Relay optical system, 10 ... Observation lens tube, 11 ... Eyepiece DESCRIPTION OF SYMBOLS 12 ... Projection tube, 13 ... Light source, 14 ... Collector lens 15 ... Cube turret, 16.17 ... Cube unit 16a ... Dichroic mirror, 17a ... Total reflection mirror 17b ... Ultraviolet cut filter, 18 ... 1st position detection means 18a ... Magnet, 18b ... Magnetic sensor 19 ... Rotating disk, 19a ... Rotating shaft 20 ... Imaging lens, 21 ... Imaging lens, 22 ... CCD camera 22a ... Imaging surface, 23 ... Cube turret 24.25 ... Cube unit, 24a ... Total reflection mirror 25a ... Dichroic mirror, 26 ... Second position detection means 26a ... Magnet, 26 b ... Magnetic sensor 27 ... Shutter, 28 ... Drive controller, 31 ... Laser light source 32 ... Mirror, 33 ... Dichroic mirror 34 ... Two-dimensional scanning mechanism, 35 ... Cube unit 36 ... Confocal pinhole, 37 ... Photo detector

Claims (10)

A microscope main body having an objective lens arranged close to the specimen, and an observation means for observing the optical image of the specimen through the objective lens;
A light source provided on the side surface of the microscope main body, and a light projecting tube having at least one imaging means;
A wavelength selection optical element that reflects light from the light source to the specimen and transmits light from the specimen to the observation means and a reflection optical element that totally reflects light from the light source to the specimen are selectively inserted into the optical path. First optical element switching means for
A wavelength selection optical element that is in the light projecting tube and reflects light from the light source to the first optical element switching means and transmits light from the sample to the imaging means, and light from the light source to the first light source. Second optical element switching means for selectively inserting a reflective optical element for total reflection into the optical path of one optical element;
A light control means for shielding or dimming light from the light source,
A microscope apparatus characterized in that, when the reflective optical elements of the first and second optical element switching means are simultaneously inserted into an optical path, the light of the light source is blocked or dimmed by the dimming means. 2. The microscope according to claim 1, wherein the light projecting tube is provided so that light from the light source is collected and a rotating disk having a plurality of through holes at positions conjugate with the sample is detachable. apparatus. The projection tube further includes a laser light source, a two-dimensional scanning unit that two-dimensionally scans the sample with laser light from the laser light source, and a confocal observation unit that captures light from the sample. The microscope apparatus according to claim 1. The first and second optical element switching means include first and second position detection means for detecting that the respective reflective optical elements are inserted in the optical path,
4. When the first and second position detecting means detect the reflective optical element inserted into the optical path at the same time, the light control means blocks or reduces the light of the light source. The microscope apparatus according to any one of the above. 2. The microscope apparatus according to claim 1, wherein the second optical element switching means is provided with an ultraviolet cut filter inserted into the optical path together with the reflective optical element, instead of the light control means. When the reflective optical elements of the first and second optical element switching means are simultaneously inserted into the optical path, either one of the first and second optical element switching means is forcibly removed from the optical path. The microscope apparatus according to claim 1, wherein the microscope apparatus is retracted. The microscope apparatus according to claim 1, wherein the light control unit includes a shutter or an ND filter. A microscope main body having an objective lens arranged close to the specimen, and an observation means for observing the optical image of the specimen through the objective lens;
A light source provided on the side surface of the microscope main body, and a light projecting tube having at least one imaging means;
A wavelength selection optical element that reflects light from the light source to the specimen and transmits light from the specimen to the observation means and a reflection optical element that totally reflects light from the light source to the specimen are selectively inserted into the optical path. First optical element switching means for
A wavelength selection optical element that is in the light projecting tube and reflects light from the light source to the first optical element switching means and transmits light from the sample to the imaging means, and light from the light source to the first light source. Second optical element switching means for selectively inserting a reflective optical element for total reflection into the optical path of one optical element;
A light control means for shielding or dimming light from the light source,
The first and second optical element switching means have first and second position detecting means for detecting that the respective reflective optical elements are inserted in the optical path, and these first and second positions are detected. A microscope apparatus characterized in that an alarm is issued when the detecting means simultaneously detects that the reflective optical element is inserted in the optical path. A microscope main body having an objective lens arranged close to the specimen, and an observation means for observing the optical image of the specimen through the objective lens;
Two-dimensional scanning that is provided on the side surface of the microscope main body and that two-dimensionally scans the sample with a first light source used for observation by the observation means and a laser beam output from a second light source different from the first light source And a light projection tube having confocal observation means for capturing light from the specimen scanned by the laser light,
A wavelength selection optical element that reflects light from the first light source to the specimen and transmits light from the specimen to the observation means, and a reflection optical element that totally reflects laser light from the second light source to the specimen. First optical element switching means for selectively inserting into the optical path;
One of the hole and the reflection optical element is in the light projecting tube so as to guide one of the light from the first light source and the laser light from the second light source to the first optical element switching means. Second optical element switching means for selectively inserting into the optical path;
A light control means for shielding or dimming light from the light source,
The reflection optical element in the first optical element switching means, and the hole or reflection optical element in the second optical element switching means, wherein light from the first light source is switched to the first optical element switching means. A microscope apparatus characterized in that when the light to be guided to the means is simultaneously inserted in the optical path, the light of the first light source is blocked or dimmed by the light control means. The second optical element switching means causes the light from the first light source to be reflected and guided to the first optical element switching means by inserting the reflective optical element into the optical path, and the vacancy hole is 10. The microscope apparatus according to claim 9, wherein the microscope apparatus guides the laser light from the second light source through the first optical element switching means by being inserted into an optical path.

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