JP2009116054A - Optical device and microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To make light incident on a desired position of a specimen. <P>SOLUTION: The microscope 11 includes a turning mirror 46 provided between an observation camera 29 for imaging a specimen 22 and an objective lens 24 to change the optical path of observation light from the specimen 22 by turning a reflecting surface thereof. The microscope 11 further includes a diaphragm 42 for passing excitation light through a conjugate position with the observation camera 29. The excitation light emitted from a fluorescence exciting light source 28 is transmitted by a dichromic mirror 44 through the diaphragm 42, thereafter traveled in the optical path of observation light reversely from the observation light, and incident on the specimen 22. Accordingly, even if the turning mirror 46 is turned to move the visual field of the observation camera 29, the position to be irradiated with the excitation light on the specimen 22 is substantially matched with the position of the visual field of the observation camera 29. The present invention can be applied to a microscope. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to an optical apparatus and a microscope that can irradiate light on a desired position of a specimen.

  Conventionally, there has been proposed a microscope that moves the position of the field of the microscope with respect to the specimen without changing the objective lens to one having a different magnification or moving the objective lens or stage of the microscope (see, for example, Patent Document 1). ). Such a microscope is provided with a rotating mirror that changes the optical path of the observation light from the specimen between the objective lens and the camera that captures the image of the specimen. Any part of the specimen can be observed simply by rotating the mirror.

JP 2005-321657 A

  However, in the above-described technique, when the specimen has to be irradiated with predetermined irradiation light such as excitation light or laser for light stimulation, it has been impossible to irradiate only the desired portion of the specimen with irradiation light.

  For example, when irradiating a sample with excitation light as irradiation light, the objective lens and the stage remain fixed. Therefore, the excitation light emitted from the excitation light source is collected by the objective lens and is always at the center of the sample, that is, the objective. The light is irradiated around the position where the optical axis of the lens and the sample intersect. Therefore, there is a difference between the position of the region observed in the microscope field on the specimen and the position of the area irradiated with the excitation light, and the excitation light is irradiated even on unnecessary parts of the specimen. There was a factor such as fading of the whole specimen.

  The present invention has been made in view of such a situation, and allows light to be irradiated only on a desired portion of a specimen.

  An optical device according to one aspect of the present invention transmits an illumination light source, an objective lens, and illumination light from the illumination light source, and causes the illumination light to enter the objective lens to irradiate a specimen with the illumination light. In addition, a half mirror that reflects the observation light incident from the objective lens and incident on the imaging unit, and a position of the image of the observation light incident on the imaging unit within the light receiving surface of the imaging unit are reflected. Changing the optical path of the observation light so as to move, and changing means for changing the optical path of the irradiation light so that the position irradiated with the irradiation light on the specimen moves. An illumination optical system connecting the light source, the changing means, and the objective lens; and an observation optical system connecting the objective lens, the changing means, and the imaging means, and the illumination optical system and a part of the observation optical system are shared. Characterized in that such an optical system.

  According to one aspect of the present invention, a sample can be irradiated with light. In particular, according to one aspect of the present invention, it is possible to irradiate light only on a desired position of a specimen.

  Embodiments of the present invention will be described below. Correspondences between the constituent elements of the present invention and the embodiments described in the specification or the drawings are exemplified as follows. This description is intended to confirm that the embodiments supporting the present invention are described in the specification or the drawings. Therefore, even if there is an embodiment which is described in the specification or the drawings but is not described here as an embodiment corresponding to the constituent elements of the present invention, that is not the case. It does not mean that the form does not correspond to the constituent requirements. Conversely, even if an embodiment is described herein as corresponding to a configuration requirement, that means that the embodiment does not correspond to a configuration requirement other than the configuration requirement. It's not something to do.

  The optical device according to the present invention transmits an illumination light source (for example, the fluorescence excitation light source 28 in FIG. 1), an objective lens (for example, the objective lens 24 in FIG. 1), and the irradiation light from the illumination light source, and a specimen. In order to irradiate the irradiation light to the objective lens, the irradiation light is incident on the objective lens, and the observation light incident on the objective lens is reflected from the specimen, thereby imaging means (for example, the observation camera 29 in FIG. 1). And the optical path of the observation light so that the position of the image of the observation light incident on the image pickup means within the light receiving surface of the image pickup means moves. Changing means for changing the optical path of the irradiation light so that the position irradiated with the irradiation light on the specimen moves (for example, the rotating mirror 46 in FIG. 1, the moving unit 2 in FIG. 10). 3 or a stepping motor 314) of FIG. 13, and an illumination optical system that connects the illumination light source, the changing means, and the objective lens, and an observation optical system that connects the objective lens, the changing means, and the imaging means And the illumination optical system and a part of the observation optical system are a common optical system.

  In the present embodiment, there is one half mirror, but a dichroic mirror that reflects observation light of different wavelengths may be provided in the same optical path and guided to different imaging means.

  In the optical apparatus, the irradiation light applied to the specimen placed on the stage of the microscope is allowed to pass, and the adjusting means for adjusting the thickness of the light beam of the irradiation light to pass (for example, the stop 42 in FIG. 1). ) Can be further provided.

  The changing unit is disposed between the objective lens and the half mirror, reflects the irradiation light on a reflection surface and enters the objective lens, and reflects the observation light on the reflection surface to reflect the half light. It is possible to change the optical path of the irradiation light and the observation light (for example, the rotating mirror 46 in FIG. 1) by making the light incident on a mirror and rotating the reflecting surface.

  The optical device is disposed between the objective lens and the half mirror, and enlarges or reduces the size of the image of the irradiation light irradiated on the specimen, and the observation light incident on the imaging means Further, zooming means (for example, zooming optical system 45 in FIG. 1) for enlarging or reducing the size of the image can be provided.

  The change means changes the optical paths of the irradiation light and the observation light by moving the lens, the half mirror, and the adjustment means constituting the magnification changing means in a predetermined direction (for example, FIG. 10). Moving part 283).

  The changing means includes a moving means (for example, FIG. 13) that changes the optical path of the irradiation light and the observation light by moving a lens constituting the zooming means in a direction perpendicular to the optical axis of the lens. The stepping motor 314), the objective lens, and the zooming unit are arranged so that the irradiation light is reflected on the reflection surface and incident on the objective lens, and the observation light is reflected on the reflection surface. Rotating reflecting means (for example, the rotating mirror 46 in FIG. 13) is provided that changes the optical paths of the irradiation light and the observation light by reflecting the light so as to be incident on the half mirror and rotating the reflecting surface, for example. be able to.

  In the present invention, the observation light incident on the changing means and the reflected observation light, and the optical paths of the irradiation light incident on the changing means and the reflected irradiation light are common.

  Embodiments to which the present invention is applied will be described below with reference to the drawings.

  FIG. 1 is a diagram showing a configuration example of an embodiment of a microscope to which the present invention is applied.

  The microscope 11 is provided with a stage 21, and a specimen 22 to be observed is arranged on the stage 21. Further, the microscope 11 is provided with a transmission illumination light source 23 for observing the specimen 22 in a bright field, and illumination light emitted from the transmission illumination light source 23 passes through an optical system (not shown) through the stage 21. In the figure, the sample 22 is irradiated from below. The illumination light applied to the specimen 22 passes through the specimen 22 and enters the eyepiece lens 26 and the observation camera 27 via the objective lens 24 and the intermediate unit 25 provided at a position facing the specimen 22.

  The intermediate unit 25 is equipped with a fluorescence excitation light source 28 for irradiating the specimen 22 with excitation light during fluorescence observation, and an observation camera 29 for capturing an image of the specimen 22. In addition, the intermediate unit 25 includes a lens 41, a diaphragm 42, a lens 43, a dichroic mirror 44, a zooming optical system 45, and a rotating mirror 46.

  The intermediate unit 25 may be provided integrally with the microscope 11 or may be a device different from the microscope 11 and detachable from the microscope 11.

  The fluorescence excitation light source 28 emits excitation light that is parallel light and makes it incident on the lens 41 of the intermediate unit 25. The lens 41 collects the excitation light incident from the fluorescence excitation light source 28 and causes the excitation light to enter the lens 43 through the diaphragm 42. Here, the lens 41 condenses the excitation light so that the excitation light forms an image at the position of the opening of the stop 42.

  The diaphragm 42 allows part or all of the excitation light incident from the lens 41 to pass through and enter the lens 43. That is, the diaphragm 42 adjusts the thickness (light quantity) of the excitation light beam passing through the aperture by appropriately changing the size of the aperture provided in the diaphragm 42. The stop 42 is disposed at a position conjugate with the observation camera 29, and the size of the opening of the stop 42 is the same as the size of the light receiving surface of the observation camera 29 (more specifically, approximately the same size). The

  The lens 43 converts the excitation light incident from the lens 41 into parallel light, and causes the excitation light converted into parallel light to enter the dichroic mirror 44. The dichroic mirror 44 transmits the excitation light incident from the lens 43 and causes the excitation light to enter the objective lens 24 via the variable magnification optical system 45 and the rotating mirror 46. The objective lens 24 collects the excitation light incident from the rotating mirror 46 and irradiates the specimen 22 with the excitation light. Note that. The thickness of the light beam of the excitation light is changed in the variable magnification optical system 45, and the size of the image of the excitation light irradiated on the specimen 22 is enlarged or reduced.

  When the sample 22 is irradiated with excitation light, the sample 22 expresses fluorescence by the irradiated excitation light. The fluorescence expressed from the specimen 22 becomes observation light for observing the specimen 22 and enters the rotating mirror 46 through the objective lens 24. The rotating mirror 46 transmits part of the observation light (fluorescence) incident from the objective lens 24 and reflects part of the incident observation light to be incident on the variable magnification optical system 45.

  The observation light transmitted through the rotating mirror 46 enters the eyepiece lens 26 and the observation camera 27 through an optical system (not shown). The eyepiece lens 26 focuses the observation light incident from the rotating mirror 46 to form an image. The observation camera 27 captures an image of the observation light incident from the rotating mirror 46 and supplies the captured image to a device (not shown) for display. Therefore, the user who is an observer can observe the specimen 22 from the eyepiece lens 26, and observe the specimen 22 imaged by the observation camera 27 by viewing an image displayed on a device (not shown). be able to.

  On the other hand, the observation light reflected by the rotating mirror 46 enters the dichroic mirror 44 via the variable magnification optical system 45. The thickness of the observation light beam changes in the variable magnification optical system 45. Thereby, the size of the image of the observation light (fluorescence) incident on the observation camera 29 is enlarged or reduced.

  The dichroic mirror 44 reflects the observation light incident from the variable magnification optical system 45 so as to enter the observation camera 29. The observation camera 29 images the observation light incident from the dichroic mirror 44, and the captured image is illustrated. To be displayed on the device.

  For example, as shown in FIG. 2, the variable magnification optical system 45 is a low magnification optical system 71 for reducing the size of the observation light image formed on the light receiving surface of the observation camera 29, that is, the image of the sample 22. And a high-magnification optical system 72 for enlarging. 2A shows a view of the variable power optical system 45 from the top to the bottom in FIG. 1, and FIG. 2B shows the variable power optical system 45 from the front to the back in FIG. The figure is shown.

  The low-magnification optical system 71 includes a variable power lens 81 and a variable power lens 82, and converts the size of the image of the sample 22 incident on the observation camera 29 to a size of 0.35 times, for example. That is, the magnification of the low magnification optical system 71 is 0.35 times. Further, the high-magnification optical system 72 converts the size of the image of the observation light (specimen 22) incident on the observation camera 29, for example, to four times the size. That is, the magnification of the high-magnification optical system 72 is 4 times. More specifically, the magnification of the observation light image in the observation camera 29 is determined by the product of the magnification of the low magnification optical system 71 or the high magnification optical system 72 and the magnification of the objective lens 24.

  The variable magnification optical system 45 is moved in the direction indicated by the arrow A11, that is, in the direction perpendicular to the optical path of the observation light and the optical axis of the objective lens 24. In FIG. 2, the variable magnification optical system 45 is positioned such that the low magnification optical system 71 is disposed between the rotating mirror 46 and the dichroic mirror 44. The optical axes of the variable magnification lens 81 and the variable magnification lens 82 are positioned on a straight line connecting the center of the reflection surface of the rotating mirror 46 and the center of the reflection surface of the dichroic mirror 44.

  In this state, the observation light reflected by the rotating mirror 46 is collected by the variable power lens 81, and an intermediate image is formed on the imaging surface S that is a position between the variable power lens 81 and the variable power lens 82. Then, the light is incident on the variable magnification lens 82. The observation light incident on the variable magnification lens 82 is collimated by the variable magnification lens 82 to become parallel light and is incident on the dichroic mirror 44. Thus, the user can observe the sample 22 at a low magnification by looking at the image captured by the observation camera 29.

  Further, when the user operates the microscope 11 to instruct a change in magnification, the zoom optical system 45 is configured so that the high-magnification optical system 72 is disposed between the rotating mirror 46 and the dichroic mirror 44. It moves in the middle, upward direction, that is, in the direction of arrow A11. At this time, the optical axes of the variable magnification lens 83 and the variable magnification lens 84 are positioned on a straight line connecting the center of the reflection surface of the rotating mirror 46 and the center of the reflection surface of the dichroic mirror 44.

  In this state, the observation light reflected by the rotating mirror 46 is collected by the variable power lens 83 to form an intermediate image on the image forming surface S and is incident on the variable power lens 84. The observation light incident on the variable magnification lens 84 is collimated by the variable magnification lens 84 to become parallel light and is incident on the dichroic mirror 44. Thereby, the user can observe the sample 22 at a high magnification by viewing the image captured by the observation camera 29.

  Incidentally, as shown in FIG. 3, the rotation mirror 46 is provided with a rotation control unit 101, and the reflection surface of the rotation mirror 46 is rotated by the rotation control unit 101. Yes. In a state where the reflecting surface of the rotating mirror 46 is not rotated, the center of the reflecting surface of the rotating mirror 46 is located at the intersection of the optical axis of the objective lens 24 and the optical axis of the zoom optical system 45, A rotating mirror 46 is arranged so that the reflecting surface and the optical axis form an angle of 45 degrees.

  The dichroic mirror 44 is arranged such that the reflection surface forms an angle of 45 degrees with respect to the optical axes of the variable magnification optical system 45 and the observation camera 29, and the center of the reflection surface is the center of those optical axes. It is designed to be located at the intersection.

  The rotation control unit 101 rotates the rotation mirror 46 about a straight line passing through the center of the reflection surface of the rotation mirror 46. For example, a straight line that passes through the center of the reflecting surface and is perpendicular to the optical axis of the objective lens 24 and the optical axis of the zoom optical system 45 is defined as the x-axis. If the y-axis is a straight line that passes through the center of the reflecting surface and is parallel to the reflecting surface and perpendicular to the x-axis, the rotation control unit 101 sets the reflecting surface of the rotating mirror 46 around the x-axis or y-axis. Rotate.

When the reflection surface of the rotation mirror 46 is rotated by the rotation control unit 101, the position of the intermediate image of the observation light reflected by the rotation mirror 46 and formed by the variable magnification lens 81 is the imaging plane S. Move up. Therefore, the position where the observation light is incident on the zoom lens 82 is also moved, and thereby the position of the image of the observation light that is imaged on the light receiving surface of the observation camera 29 is also moved. Here, the observation light incident on the zoom lens 82 is reflected by the dichroic mirror 44 and enters the imaging lens 102 provided in the observation camera 29. The observation light incident on the imaging lens 102 is collected by the imaging lens 102 and an image of the observation light is formed on the light receiving surface of the observation camera 29.

  As described above, the specimen 22 is arranged by rotating the reflecting surface of the rotating mirror 46 to change the optical path of the observation light and moving the intermediate image formed by the variable magnification lens 81 on the imaging plane S. This is equivalent to moving the stage 21 in a direction perpendicular to the optical axis of the objective lens 24.

  Therefore, as shown in FIG. 4, when the rotating mirror 46 is rotated about the x axis and the intermediate image is moved in the vertical direction in the figure, the field of view of the image captured by the observation camera 29 is the specimen 22. Move in the left-right direction in the figure above. Further, when the rotating mirror 46 is rotated about the y-axis to move the intermediate image in the x-axis direction in the figure, the field of view of the image captured by the observation camera 29 is x in the figure on the specimen 22. Move in the axial direction.

  For example, as shown in FIG. 5, when a plurality of cultured cells as the specimen 22 are observed in the field of view of the microscope 11 at a low magnification, the user moves the zoom optical system 45 and rotates it appropriately. By rotating the mirror 46, any cultured cells in the field of view can be observed at a high magnification.

  In FIG. 5, a rectangular area 131 indicates the field of view of the microscope 11, that is, the field of view of the observation camera 29 when the low-magnification optical system 71 is located on the optical path of the observation light. Each of the rectangular regions 132-1 to 132-5 represents the field of view of the microscope 11 (observation camera 29) when the high-magnification optical system 72 is located on the optical path of the observation light. That is, the region 131 shows the field of view of the microscope 11 at a low magnification, and the regions 132-1 to 132-5 show the field of view of the microscope 11 at a high magnification.

  When the user observes the cultured cell as the specimen 22 at a low magnification and finds a cultured cell 133 that fits the purpose of observation in the field of view, the magnification changing optical system 45 is moved to switch the magnification to a high magnification. The rotating mirror 46 is rotated. Then, for example, the region 132-1 on the specimen 22 is observed in the visual field of the microscope 11. The user further rotates the rotating mirror 46 to move the position of the visual field on the specimen 22 to the region 132-2 to the region 132-5 and the like. The field of view is moved to the region 132-3 observed in FIG. As a result, the cultured cells 133 can be observed within the field of view of the microscope 11 as shown on the right side in the figure.

  As described above, in the microscope 11, the visual field of the microscope 11 can be moved only by rotating the rotating mirror 46 without moving the stage 21 and the objective lens 24.

  In the microscope 11, the diaphragm 42 is disposed at a position conjugate with the light receiving surface of the observation camera 29, and the excitation light image (intermediate image) emitted from the fluorescence excitation light source 28 is the position of the opening of the diaphragm 42. Formed. Therefore, when the rotation mirror 46 is rotated, not only the field of view of the observation camera 29 (the microscope 11) is moved, but also the region where the sample 22 is irradiated with the excitation light is moved. That is, for example, as shown in FIG. 6, in the region irradiated with the excitation light on the specimen 22, the center position is the same as the center position of the area that becomes the field of view of the observation camera 29 on the specimen 22. The region is almost the same size as the region serving as the field of view of the observation camera 29.

  In FIG. 6, a circular region 161 indicates a region irradiated with excitation light on the specimen 22, and a rectangular region 162 indicates a region that is a field of view of the observation camera 29 at a low magnification. Yes. A rectangular region 163 indicates a region that is a field of view of the observation camera 29 at a high magnification.

  When the user observes the sample 22 at a low magnification, that is, when the low-magnification optical system 71 is arranged on the optical path of the observation light, the region 162 is observed in the field of view of the observation camera 29. When the user is observing the specimen 22 at a low magnification, the excitation light from the fluorescence excitation light source 28 is irradiated to the entire region 161. Thus, in the microscope 11, the field 162 of the field of view of the observation camera 29 and the area 161 irradiated with the excitation light are substantially the same area.

  Further, when the user switches the magnification of the microscope 11 from the low magnification to the high magnification and appropriately rotates the rotating mirror 46, the field of view of the observation camera 29 moves from the region 162 to the region 163. That is, when the microscope 11 is operated so that the high-magnification optical system 72 is disposed on the optical path of the observation light, the region 163 is observed in the field of view of the observation camera 29.

  Here, in the conventional microscope, the excitation light from the fluorescence excitation light source is disposed between the parallel optical system immediately after the objective lens, for example, between the objective lens and the rotating mirror. Regardless of the position, the range where the excitation light on the specimen is irradiated is always a range centered on the optical axis of the objective lens. Therefore, when the center of the field of view of the observation camera 29 is different from the intersection with the optical axis of the objective lens on the specimen, there is a deviation between the region irradiated with the excitation light and the region used as the field of view of the observation camera. As a result, the specimen was irradiated with excitation light in vain.

  On the other hand, in the microscope 11, the center of the region irradiated with the excitation light on the specimen 22 and the field of view of the observation camera 29 are used regardless of the magnification at the time of observation and the movement of the field of view of the observation camera 29. Since it is always at the same position as the center of the region, the sample 22 is not unnecessarily irradiated with excitation light.

  For example, when the magnification is switched to a high magnification and the field of view of the observation camera 29 is moved to the region 163, the region irradiated with the excitation light is the region 163 of the field of view of the observation camera 29 as shown in FIG. It becomes almost the same area. In FIG. 7, parts corresponding to those in FIG. 6 are denoted by the same reference numerals, and description thereof is omitted.

  In FIG. 7, a region 191 indicates a region on the specimen 22 that is irradiated with excitation light. The region 191 includes a visual field region 163, and the center thereof is a circular region having the same position as the center of the region 163.

  When the magnification of the variable magnification optical system 45 is switched to a high magnification, the excitation light emitted from the fluorescence excitation light source 28 enters the variable magnification optical system 45 via the lens 41 or the dichroic mirror 44. Then, the variable magnification optical system 45 changes the thickness of the luminous flux of the excitation light so that the area irradiated with the excitation light in the specimen 22 is almost the same size as the field of view of the observation camera 29. The light passes through the optical path of the observation light in the direction opposite to that of the observation light and is applied to the specimen 22. That is, the excitation light emitted from the variable magnification optical system 45 is reflected by the rotating mirror 46 and enters the objective lens 24. Then, the excitation light is condensed by the objective lens 24 and irradiated on the specimen 22. Here, when the rotating mirror 46 is rotated, the optical path of the excitation light is changed together with the optical path of the observation light.

  In the microscope 11, the diaphragm 42 and the observation camera 29 are in a conjugate relationship, and the aperture of the diaphragm 42 is the same size as the light receiving surface of the observation camera 29. Therefore, when the excitation light is irradiated from the dichroic mirror 44 through the optical path of the observation light in the opposite direction to the specimen 22, the area irradiated with the excitation light on the specimen 22 is always substantially the same as the visual field of the observation camera 29. It becomes. Of course, the user can change the size of the region where the sample 22 is irradiated with the excitation light by operating the microscope 11 and changing the size of the aperture of the diaphragm 42.

  Further, when the laser light source is used, the diaphragm 42 is not necessarily required because the laser light itself is already focused light.

  In this way, the excitation light is always applied to a region substantially the same as the region of the field of view of the observation camera 29 on the specimen 22. Here, when the user performs fluorescence observation, the region of the field of view of the observation camera 29 is the region that the user pays attention to and observes. Therefore, the region is irradiated with excitation light for the user to perform fluorescence observation. The desired region, that is, the region desired by the user. Therefore, by providing the dichroic mirror 44 between the rotating mirror 46 and the observation camera 29 and providing the stop 42 at a position conjugate with the observation camera 29, the excitation light is irradiated only on the region desired by the user on the specimen 22. Can be made. Thereby, the sample 22 can be observed stably for a long time.

  Since the microscope 11 is provided with the rotating mirror 46 whose reflecting surface rotates, the field of view of the observation camera 29 can be moved without moving the objective lens 24 and the stage 21.

  Furthermore, the microscope 11 described above can be applied to an observation system for fluorescent observation of the specimen 22 as shown in FIG. 8, for example. The observation system shown in FIG. 8 includes a microscope 11, a computer 221, a mirror controller 222, and an observation monitor 223.

  For example, the user operates the computer 221, the microscope 11, and the fluorescence excitation light source 28 as necessary to observe the specimen 22 in fluorescence. When the user operates the fluorescence excitation light source 28 to emit excitation light, the computer 221 controls the observation camera 29 to capture an image of the specimen 22 and acquires an image captured by the observation camera 29. Then, the computer 221 supplies the acquired image to the observation monitor 223 for display.

  When the image of the specimen 22 is displayed on the observation monitor 223 in this way, the user sets the computer 221 so that the desired position of the specimen 22 is displayed on the observation monitor 223 while viewing the displayed image. Manipulate. The computer 221 instructs the mirror controller 222 to rotate the rotation mirror 46 according to the user's operation, and the mirror controller 222 controls the rotation control unit 101 according to the instruction from the computer 221 to turn the rotation mirror. The reflecting surface 46 is rotated.

  As a result, the position of the field of view of the observation camera 29 on the specimen 22 is moved, and the excitation light is irradiated to the area substantially the same as the area of the field of view, and the user observes a desired portion of the specimen 22 on the observation monitor 223. can do. Note that the computer 221 may control the microscope 11 in accordance with a user operation and move the zoom optical system 45 to change the magnification.

  Further, for example, as shown in FIG. 9, a manipulator 251 for performing a predetermined operation such as applying an electrical stimulus to the specimen 22 in the observation system and a manipulator controller 252 may be further provided. In FIG. 9, the same reference numerals are given to the portions corresponding to those in FIG. 8, and the description thereof is omitted.

  In the observation system of FIG. 9, the user operates the computer 221 while viewing the image of the specimen 22 displayed on the observation monitor 223 to move the position of the visual field of the observation camera 29. Then, the computer 221 instructs the mirror controller 222 to rotate the rotation mirror 46 and instructs the manipulator controller 252 to operate the manipulator 251 based on the rotation angle of the rotation mirror 46.

  The manipulator controller 252 controls the operation of the manipulator 251 in response to an instruction from the computer 221. That is, the manipulator controller 252 moves the tip of the electrode needle provided on the manipulator 251 to the movement destination area of the field of view of the observation camera 29 on the specimen 22, and performs a predetermined operation on the specimen 22 on the manipulator 251. Make it. As described above, the computer 221 instructs the manipulator controller 252 to operate the manipulator 251 in accordance with the movement of the field of view of the observation camera 29, so that the user can observe the sample 22 with a simple operation.

  In the observation system shown in FIG. 9 also, when the position of the field of view of the observation camera 29 on the specimen 22 is moved, the excitation light is also irradiated onto the substantially same region as the region of the field of view. The desired portion of the specimen 22 can be observed at. Furthermore, since the manipulator 251 is moved in accordance with the movement of the visual field of the observation camera 29, it is not necessary for the user to move the manipulator 251 directly.

  Further, in the above description, it has been described that the field of view of the observation camera 29 is moved by rotating the rotation mirror 46. However, the field of view of the observation camera 29 is moved by moving other optical elements provided in the intermediate unit 25. May be moved.

  In such a case, the intermediate unit 25 includes a housing 281 and a housing 282 as shown in FIG. 10A corresponds to FIG. 2A and shows a view of the variable magnification optical system 45 from the top to the bottom in FIG. FIG. 10B corresponds to FIG. 2B, and shows a view of the variable magnification optical system 45 from the near side to the far side in FIG. Further, in FIG. 10, the same reference numerals are given to the portions corresponding to those in FIG. 2, and the description thereof is omitted.

  In FIG. 10, a lens 41, a diaphragm 42, a lens 43, a dichroic mirror 44, a variable power lens 82, and a variable power lens 84 are provided inside the housing 281. In addition, an observation camera 29 and a fluorescence excitation light source 28 (not shown) are attached to the outside of the housing 281, and a moving unit 283 that moves the housing 281 relative to the housing 282 is provided.

  For example, assuming that the direction perpendicular to the optical axis of the objective lens 24 and the optical axis of the variable magnification optical system 45 is the x direction and the direction parallel to the optical axis of the objective lens 24 is the y direction, the moving unit 283 has a housing 281 is moved in the x and y directions. Therefore, when the housing 281 moves, the lens 41, the diaphragm 42, the lens 43, the dichroic mirror 44, the zoom lens 82, and the zoom lens 84 in the housing 281 also move in the x direction and the y direction. In addition, the observation camera 29 and the fluorescence excitation light source 28 mounted on the housing 281 also move in the x direction and the y direction.

  On the other hand, the housing 282 is not moved, and the housing 282 is provided with a zoom lens 81, a zoom lens 83, and a fixed mirror 284. The center of the reflecting surface of the fixed mirror 284 is located at the intersection of the optical axis of the objective lens 24 and the optical axis of the variable magnification optical system 45, and the reflecting surface and those optical axes form an angle of 45 degrees. Is arranged. The reflecting surface of the fixed mirror 284 is fixed so as not to rotate.

  Further, in the intermediate unit 25 of FIG. 10, the zoom optical system 45 is moved in the x direction independently of the movement of the housing 281 by the moving unit 283. That is, the variable magnification optical system 45 moves separately from the housing 281 so that the low magnification optical system 71 or the high magnification optical system 72 is positioned on the optical path of the observation light.

  Further, for example, as shown in FIG. 11, when the observation light, which is fluorescence generated from the specimen 22, enters the fixed mirror 284 from the objective lens 24 from the bottom to the top in the drawing, the fixed mirror 284 transmits the observation light. The light is reflected and incident on the variable magnification lens 81. The observation light incident on the variable magnification lens 81 from the fixed mirror 284 is collected by the variable magnification lens 81 and forms an intermediate image on the image plane S. Thereafter, the observation light is incident on the light receiving surface of the observation camera 29 via the variable power lens 82 dichroic mirror 44 and the imaging lens 102, and an image of the observation light is formed.

  Here, for example, the moving unit 283 moves the housing 281 in the x direction and the y direction in a state where the light beam forming the central portion of the intermediate image of the observation light is incident on the variable magnification lens 82 among the light beams of the observation light. Suppose. In this case, after the housing 281 is moved, the zoom lens 81 to the zoom lens 82 are moved in the x direction and the y direction from the center of the intermediate image of the observation light on the imaging surface S of the observation light flux. A light beam forming a shifted portion is incident.

  In other words, the zoom lens 82 and the dichroic mirror 44 are moved by the movement of the housing 281, so that the optical path of the observation light is changed, and the position of the image of the observation light on the light receiving surface of the observation camera 29 is also moved. This is equivalent to moving the stage 21 on which the specimen 22 is arranged in a direction perpendicular to the optical axis of the objective lens 24.

  Therefore, when the housing 281 is moved in the x direction, the field of view of the image captured by the observation camera 29 moves in the x direction on the sample 22 without moving the stage. When the housing 281 is moved in the y direction, the field of view of the image captured by the observation camera 29 is also moved in the left-right direction on the sample 22 without moving the stage.

  Further, since the fluorescence excitation light source 28 is also moved together with the housing 281, the excitation light emitted from the fluorescence excitation light source 28 matches the field of view of the observation camera 29 in accordance with the movement of the field of view of the observation camera 29. Irradiates almost the same area. That is, the excitation light emitted from the fluorescence excitation light source 28 enters the dichroic mirror 44 through the lenses 41 to 43 and passes through the dichroic mirror 44. The excitation light that has passed through the dichroic mirror 44 is then irradiated on the specimen 22 through the optical path of the observation light in the direction opposite to the observation light. Therefore, when the housing 281 is moved, the optical path of the excitation light is changed, the visual field is moved, and the position where the excitation light is irradiated on the specimen 22 is also moved.

  In this manner, even when the housing 281 is moved to move the field of view of the observation camera 29, the excitation light can be irradiated only on the region on the specimen 22 that is substantially the same as the field of view desired by the user. Even when the variable magnification optical system 45 is moved and the high magnification optical system 72 is arranged on the optical path of the observation light, as in the case of the low magnification optical system 71, the excitation light is observed on the observation camera on the specimen 22. It irradiates almost the same area as 29 fields of view.

  Further, in order to move the field of view of the observation camera 29, the variable power lens 81 or the variable power lens 83 constituting the variable power optical system 45 may be moved. In such a case, when the low magnification optical system 71 is arranged on the optical path of the observation light, the variable magnification lens 81 is moved, and when the high magnification optical system 72 is arranged on the optical path of the observation light, the variable magnification lens 83 is moved. Is moved. As described above, by moving the variable magnification lens 81 or the variable magnification lens 83, the optical paths of the observation light and the excitation light are changed as in the case of FIG.

  For example, when the zoom lens 81 is moved, the zoom lens 81 is moved in a direction perpendicular to the optical axis of the zoom optical system 45 and the optical axis of the objective lens 24 as shown in FIG. FIG. 12 is a view of the microscope 11 of FIG. 1 as viewed from above.

  In FIG. 12, when the variable magnification lens 81 is moved in a direction perpendicular to the optical axis of the variable magnification optical system 45 and the optical axis of the objective lens 24, that is, in the vertical direction in the figure, the variable magnification lens 81 is formed. The intermediate image of the observation light is moved up and down. Therefore, even when the zoom lens 81 is moved in the vertical direction, the optical path of the observation light is changed and the field of view of the observation camera 29 is moved in the vertical direction in the drawing on the sample 22 as in FIG. . When the zoom lens 81 is moved in the vertical direction, the optical path of the excitation light is also changed, and the position on the sample 22 where the excitation light is irradiated is also moved in the vertical direction.

  Further, when the variable magnification lens 81 is moved in the vertical direction, the rotating mirror 46 makes a straight line that passes through the center of the reflecting surface and is perpendicular to the optical axis of the variable magnification optical system 45 and the optical axis of the objective lens 24. It rotates as an axis. When the rotating mirror 46 rotates, the intermediate image of the observation light formed on the imaging surface S moves in a direction parallel to the optical axis of the objective lens 24, so that the field of view of the observation camera 29 is on the specimen 22. In the figure, move left and right. Further, the optical path of the excitation light is changed by the rotation of the rotation mirror 46, and the position where the excitation light is irradiated on the specimen 22 also moves in the left-right direction.

  The zoom lens 81 is moved in the vertical direction and in a direction parallel to the optical axis of the objective lens 24 so that the field of view of the observation camera 29 is moved in the vertical direction and the horizontal direction in the drawing on the sample 22. May be.

  Further, the zoom lens 81 is moved in a direction parallel to the optical axis of the objective lens 24, the field of view of the observation camera 29 is moved in the left-right direction in the drawing on the sample 22, and the rotating mirror 46 is rotated. Thus, the visual field of the observation camera 29 may be moved in the vertical direction on the specimen 22. In such a case, a straight line that is perpendicular to the optical axis of both the optical axis of the zoom optical system 45 and the optical axis of the objective lens 24 and that passes through the center of the reflecting surface of the rotating mirror 46 is used as an axis. The rotating mirror 46 is rotated.

  Further, when the zoom lens 81 is moved in the vertical direction, for example, as shown in FIG. 13, the zoom lens 81 is moved by a stepping motor 314.

  In FIG. 13, the zoom lens 81 is fixed by a lens holder 311, and the lens holder 311 is disposed on the guide rail 312. Further, the lower end of the guide rail 312 and the lens holder 311 are connected by a spring 313-1 and a spring 313-2 in the drawing. Further, a stepping motor 314 is disposed at a position adjacent to the lens holder 311 between the spring 313-1 and the spring 313-2.

  For example, when the user operates the microscope 11 so that the zoom lens 81 is moved, the stepping motor 314 moves the shaft 315 provided at the center thereof in the vertical direction. For example, when the stepping motor 314 moves the shaft 315 upward in the drawing, the lens holder 311 is pushed upward by the shaft 315 and moves upward on the guide rail 312. Further, for example, when the stepping motor 314 moves the shaft 315 downward, the spring 313-1 and the spring 313-2 contract, so that the lens holder 311 is pulled downward by the spring 313-1 and the spring 313-2. The guide rail 312 is moved downward.

  In this way, when the stepping motor 314 is driven and the lens holder 311 moves up and down on the guide rail 312, the zoom lens 81 fixed to the lens holder 311 also moves. That is, the variable magnification lens 81 is moved in a direction perpendicular to the optical axis of the variable magnification lens 81 by the stepping motor 314. When the zoom lens 81 moves in the vertical direction, the intermediate image of the observation light formed by the zoom lens 81 moves in the vertical direction on the imaging surface S.

  Furthermore, the guide rail 312 is provided with an LED (Light Emitting Diode) type position sensor 316 that detects the position of the lens holder 311. The LED type position sensor 316 is provided with a groove (not shown). When the position of the lens holder 311 is moved to a predetermined position on the upper side in the drawing, the lens is placed in the groove of the LED type position sensor 316. A part of the holder 311 is made to reach. Here, the position of the lens holder 311 when a part of the lens holder 311 reaches the groove is, for example, when the variable magnification lens 81 moves further upward, the variable magnification lens 81 forms an intermediate image of the observation light. It will be in a position where it will be impossible.

  When the LED type position sensor 316 detects that the lens holder 311 has reached the groove, the stepping motor 314 moves the shaft 315 downward in the figure by a predetermined number of steps, thereby changing the magnification. The lens 81 is also moved downward.

  In the above description, the variable magnification optical system 45 is provided in the intermediate unit 25. However, the variable magnification optical system 45 may not be provided. Even in such a case, the field of view of the observation camera 29 is moved by the rotation of the rotation mirror 46, and the excitation light from the fluorescence excitation light source 28 is substantially the same as the field of view of the observation camera 29 on the specimen 22. Irradiate the area.

  The light irradiated on the specimen 22 is not limited to a laser light source as an excitation light, a mercury lamp, a xenon lamp, or the like. For example, a laser for light stimulation, an ultraviolet light that activates a cell as the specimen 22 or the like. It can be. In such a case, instead of the fluorescence excitation light source 28, a light source such as a laser or ultraviolet light irradiated on the specimen 22 is attached to the intermediate unit 25, and the light emitted from the light source is emitted from the lens 41 to the objective. The specimen 22 is irradiated through the lens 24. Alternatively, a light source that emits visible light may be used.

  The embodiment of the present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the gist of the present invention.

It is a figure which shows the structural example of one Embodiment of the microscope to which this invention is applied. It is a figure which shows the structural example of a variable magnification optical system. It is a figure explaining rotation of a rotation mirror. It is a figure explaining the movement of the visual field of an observation camera. It is a figure explaining the movement of the visual field of an observation camera. It is a figure explaining the area | region irradiated with excitation light. It is a figure explaining the area | region irradiated with excitation light. It is a figure which shows the structural example of an observation system. It is a figure which shows the other structural example of an observation system. It is a figure which shows the other structural example of an observation irradiation unit. It is a figure explaining the movement of an optical system. It is a figure explaining the movement of a variable magnification lens. It is a figure explaining the mechanism which moves a variable magnification lens.

Explanation of symbols

  11 microscope, 22 specimens, 24 objective lens, 25 intermediate unit, 28 light source for fluorescence excitation, 29 observation camera, 42 aperture, 44 dichroic mirror, 45 variable magnification optical system, 46 rotating mirror, 71 low magnification optical system, 72 high magnification Optical system, 81 zoom lens, 82 zoom lens, 83 zoom lens, 84 zoom lens, 101 rotation control unit, 221 computer, 222 mirror controller, 281 housing, 283 moving unit, 284 fixed mirror, 315 stepping motor

Claims (9)

An illumination light source;
An objective lens;
The irradiation light from the illumination light source is transmitted, and the irradiation light is incident on the objective lens in order to irradiate the sample with the irradiation light, and the observation light from the sample incident from the objective lens is reflected. A half mirror incident on the imaging means;
While changing the optical path of the observation light so that the position of the image of the observation light incident on the imaging means moves within the light receiving surface of the imaging means, the position where the irradiation light is irradiated on the sample is Changing means for changing the optical path of the irradiation light so as to move,
An illumination optical system that connects the illumination light source, the changing unit, and the objective lens, and an observation optical system that connects the objective lens, the changing unit, and the imaging unit, and the illumination optical system and a part of the observation optical system are An optical device characterized by being a common optical system. An optical path where irradiation light is incident on the changing means in the illumination optical system, an optical path where irradiation light is reflected by the changing means, and an optical path where observation light is incident on the changing means and the observation light in the observation optical system are reflected. The optical device according to claim 1, wherein the optical path is common. 2. The apparatus according to claim 1, further comprising an adjusting unit configured to transmit the irradiation light applied to the specimen arranged on a stage of a microscope and to adjust a thickness of a light beam of the irradiation light to be transmitted. Optical device. The optical apparatus according to claim 3, wherein an opening provided in the adjustment unit and through which the irradiation light passes is made the same size as the light receiving surface of the imaging unit. The changing unit is disposed between the objective lens and the half mirror, reflects the irradiation light on a reflection surface and enters the objective lens, and reflects the observation light on the reflection surface to reflect the half light. The optical apparatus according to any one of claims 1 to 4, wherein an optical path of the irradiation light and the observation light is changed by making the light incident on a mirror and rotating the reflection surface. The size of the image of the observation light incident between the objective lens and the half mirror, which enlarges or reduces the size of the image of the irradiation light irradiated on the specimen and enters the imaging means The optical apparatus according to claim 3, further comprising a scaling unit that expands or contracts the zoom lens. The changing means changes the optical path of the irradiation light and the observation light by moving a lens, the half mirror, and the adjustment means that constitute the magnification changing means in a predetermined direction. Item 7. The optical device according to Item 6. The changing means is
Moving means for changing the optical path of the irradiation light and the observation light by moving the lens constituting the zooming means in a direction perpendicular to the optical axis of the lens;
Arranged between the objective lens and the zooming means, the irradiation light is reflected on the reflection surface and incident on the objective lens, and the observation light is reflected on the reflection surface to the half mirror. The optical apparatus according to claim 6, further comprising: a rotating reflecting unit that changes an optical path of the irradiation light and the observation light by being incident and rotating the reflecting surface.   A microscope comprising the optical device according to claim 1.

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