JP2018066848A - Sheet illumination microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To suppress occurrence of shadow arising from illumination without upsizing a device.SOLUTION: A sheet illumination microscope 1 is provided that comprises: an illumination optical system 3 that makes a sheet-like illumination light incident with respect to a sample X housed in a sample container 2; and an observation optical system 4 that includes an observation optical axis P orthogonal to an incidence plane of the illumination light to be incident made by the illumination optical system 3, and forms an image of the sample X by a light generating in the sample X due to irradiation of the illumination light, in which the illumination optical system 3 comprises: a cylindrical light flux formation optical system 8 that forms a cylindrical surface-like light flux having a center axis parallel with the observation optical axis P; a deflection optical system 9 that deflects a part of the light flux formed by the cylindrical light flux formation optical system 8 in a direction along the incidence plane to form the illumination light; and a movement mechanism 10 that moves the deflection optical system 9 around the center axis.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a sheet illumination microscope.

2. Description of the Related Art Conventionally, a sheet illumination microscope is known that can acquire a good three-dimensional stereoscopic image of a sample at a high speed while suppressing the fading of fluorescence without irradiating illumination light to a place other than the imaging plane of the sample (for example, , Patent Document 1 and Patent Document 2).
In the sheet illumination microscope of Patent Document 1, when a part that does not easily transmit illumination light due to absorption or a part that scatters light is included in the sample, the illumination light does not enter behind these parts. In order to prevent the inconvenience that a shadow is formed in the field of view, the sample container is rotated.

  In addition, the sheet illumination microscope of Patent Document 2 scans the direction of the light beam component in the sheet-like illumination light with a galvano mirror, a polygon mirror, or the like for the purpose of reducing shadows formed in the sample due to illumination. It is changed depending on the device.

JP 2011-215644 A Japanese Patent No. 5525136

  However, in the sheet illumination microscope disclosed in Patent Document 1, when the sample is a living organism such as a zebrafish, for example, the sample container may be moved to cause stimulation or visual stimulation due to acceleration. There is a possibility of adversely affecting the observation. In addition, in the case of a sample container in which a plurality of samples are arranged in an array, a large space is required for rotating the sample container, and there is a disadvantage that the apparatus becomes large.

Further, in the sheet illumination microscope of Patent Document 2, an optical scanning device such as a galvanometer mirror or a polygon mirror is required to change the angle of the light beam component in the sheet-like illumination light. There is an inconvenience that it becomes.
The present invention has been made in view of the above-described circumstances, and an object of the present invention is to provide a sheet illumination microscope that can suppress generation of shadows due to illumination without increasing the size of the apparatus.

In order to achieve the above object, the present invention provides the following means.
One embodiment of the present invention includes an illumination optical system that makes a sheet-shaped illumination light incident on a sample contained in a sample container, and an observation optical axis that is orthogonal to an incident plane of the illumination light that is incident by the illumination optical system. An observation optical system that forms an image of the sample by light generated in the sample by irradiation of the illumination light, and the illumination optical system has a cylindrical surface shape having a central axis parallel to the observation optical axis A cylindrical light beam forming optical system for forming the light beam, a deflection optical system for deflecting a part of the light beam formed by the cylindrical light beam forming optical system in a direction along the incident plane, and forming the illumination light, and the deflection Provided is a sheet illumination microscope including a moving mechanism for moving an optical system around the central axis.

  According to this aspect, a part of the cylindrical surface light beam formed by the cylindrical light beam forming optical system is deflected by the deflection optical system, and is incident on the sample along the incident plane perpendicular to the observation optical axis. Sheet-like illumination light is formed. An image of the sample is formed by the observation optical system from light emitted from the incident plane on which the sheet-like illumination light is irradiated on the sample, and emitted from a direction along the observation optical axis perpendicular to the incident plane.

  Thereby, an image of a wide range of the sample can be observed at a time based on light generated in a wide area of the sample. Then, by moving the deflection optical system around the central axis by operating the moving mechanism, the incident angle of the illumination light is changed, so that there is a portion in the sample where light is not easily transmitted or light is scattered. However, the illumination light can be incident on the shadow area of those portions, and the generation of shadows in the observed sample image can be suppressed. That is, an optical operation device such as a galvanometer mirror or a polygon mirror is not required, and generation of shadows due to illumination can be suppressed without increasing the size of the device.

In the above aspect, the cylindrical light beam forming optical system may form a cylindrical surface light beam having a central axis substantially coinciding with the observation optical axis.
By doing so, a cylindrical light beam having the observation optical axis as the central axis is formed around the observation optical axis. As a result, one or more sheet-shaped illumination lights directed radially inward along the incident plane can be formed at an arbitrary position in the circumferential direction around the observation optical axis, and the central axis is rotated by the operation of the moving mechanism. The incident angle can be changed by moving the illumination light.

In the above aspect, the cylindrical light beam forming optical system may form the cylindrical light beam on the opposite side of the central axis across the observation optical axis.
In this way, when the illumination light is moved around the central axis, the sheet-like illumination light can be moved in the incident plane so as to cross the observation optical axis, and the illumination light is spread over a wider area of the sample. It can be made incident.

In the above aspect, the illumination optical system may include a plurality of sets of the deflection optical system and the moving mechanism with respect to a single cylindrical light beam forming optical system.
In this way, a plurality of sheet-like illumination lights are formed by a plurality of deflection optical systems from a cylindrical surface light beam formed by a single cylindrical light beam forming optical system, and are provided in each deflection optical system. The incident angle of each illumination light can be changed by the moving mechanism. By illuminating the sample from a plurality of directions, it is possible to prevent the illumination from being unbalanced by being scattered or absorbed in the sample, and to illuminate the sample uniformly.

Moreover, in the said aspect, you may provide the movable stage which moves the said sample container to the direction orthogonal to the said observation optical axis at least.
In this way, the sample container or the observation range of the sample can be changed by moving the sample container in the direction orthogonal to the observation optical axis by the operation of the movable stage.

Moreover, in the said aspect, the said sample container may be able to accommodate the said sample in a different position along the moving direction by the said movable stage.
In this way, by operating the movable stage, the sample container is moved in a direction perpendicular to the observation optical axis, and another sample accommodated in a different position is placed on the observation optical axis for observation. Can do.

Moreover, in the said aspect, the said deflection | deviation optical system may form the said illumination light which consists of a light beam which has a fixed width | variety in the said incident plane.
By doing in this way, the illumination light which consists of a sheet-like light beam which has a fixed width | variety is irradiated to a sample, and light is generated in the irradiation range of the fixed width | variety in a sample.

In the above aspect, the deflecting optical system may form a light beam that converges in a plane including the illumination optical axis of the illumination light and the observation optical axis along the incident plane.
By doing in this way, sheet-like illumination light can be formed thinly in a predetermined range substantially coinciding with the observation optical axis, and the spatial resolution of the sample image can be improved.

In the above aspect, a plurality of the deflection optical systems may be provided, and each of the deflection optical systems may form the illumination light condensed at different positions.
By doing in this way, the sheet-like illumination light formed by each deflection optical system is formed to be the thinnest at different positions, so that an image of the sample with high spatial resolution can be observed in a wider range.

In the above aspect, the deflection optical system may include an optical element having a positive power for converging the illumination light, and may be configured by a replaceable module for switching the focal length of the illumination light. Good.
By doing so, it is possible to easily observe images of samples having different spatial resolutions by exchanging the deflection optical system composed of modules configured to have different focal lengths of illumination light.

In the above aspect, a plurality of the observation optical systems may be provided.
By doing in this way, the several sample in the sample container arrange | positioned in multiple places can be observed simultaneously. Thereby, a large amount of samples can be observed at high speed.

In the above aspect, the deflecting optical system may include two or more deflecting members arranged in series.
By doing so, the cylindrical surface light beam formed by the cylindrical light beam forming optical system is deflected twice or more by two or more deflecting members arranged in series, so that the cylindrical surface light beam is relatively Even if it has a large diameter dimension, the length dimension from the observation optical axis of the sheet-like illumination light arranged in the direction orthogonal to the observation optical axis can be suppressed sufficiently small.

  According to the present invention, there is an effect that generation of shadows due to illumination can be suppressed without increasing the size of the apparatus.

It is a longitudinal section showing the whole composition of the sheet illumination microscope concerning one embodiment of the present invention. It is a top view which shows the sheet | seat illumination microscope of FIG. It is a partial longitudinal cross-sectional view which shows the 1st modification of the sheet | seat illumination microscope of FIG. It is a top view which shows the 2nd modification of the sheet illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 3rd modification of the sheet illumination microscope of FIG. It is a figure which shows the arrow A-B-C-D of the sheet | seat illumination microscope of FIG. It is a figure which shows arrow view DE of the sheet | seat illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 4th modification of the sheet | seat illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 5th modification of the sheet illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 6th modification of the sheet | seat illumination microscope of FIG. It is a top view which shows the sheet | seat illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 7th modification of the sheet illumination microscope of FIG. It is a top view which shows the sheet | seat illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 8th modification of the sheet | seat illumination microscope of FIG. It is a top view which shows the sheet | seat illumination microscope of FIG. It is a longitudinal cross-sectional view which shows the 9th modification of the sheet illumination microscope of FIG. It is a top view which shows the sheet | seat illumination microscope of FIG. FIG. 17 is a longitudinal sectional view showing another module of the deflection optical system of the sheet illumination microscope of FIG. 16. It is a longitudinal cross-sectional view which shows the 10th modification of the sheet | seat illumination microscope of FIG. FIG. 20 is a plan view showing the sheet illumination microscope of FIG. 19. It is a partial longitudinal cross-sectional view which shows the 11th modification of the sheet | seat illumination microscope of FIG.

A sheet illumination microscope 1 according to an embodiment of the present invention will be described below with reference to the drawings.
As shown in FIGS. 1 and 2, the sheet illumination microscope 1 according to the present embodiment includes an illumination optical system 3 that irradiates a sample container 2 containing a sample X such as zebrafish with sheet-like illumination light, and An observation optical system 4 for photographing light emitted from the sample X irradiated with illumination light by the illumination optical system 3, for example, fluorescence, and a movable stage 5 that supports the sample container 2 movably in the horizontal direction are provided. Yes.

  The sample container 2 includes a plurality of storage units 6 that store the sample X. By moving the sample container 2 in the horizontal direction by the operation of the movable stage 5, it is possible to change the accommodating portion 6 arranged at the position where the illumination optical system 3 forms the sheet-like illumination light. Yes.

  The illumination optical system 3 includes a beam expander (not shown). The illumination optical system 3 emits a laser beam composed of a substantially parallel beam having a predetermined beam diameter vertically upward, and the entire circumference from the laser beam emitted from the light source unit 7. A cylindrical light beam forming optical system 8 that forms a cylindrical surface light beam, and a part of the cylindrical surface light beam formed by the cylindrical light beam forming optical system 8 is deflected so as to be within the radial direction of the cylindrical light beam. A deflecting optical system 9 that forms sheet-shaped illumination light traveling in the direction and a moving mechanism 10 are provided.

  The cylindrical light beam forming optical system 8 is configured such that laser light emitted from the light source unit 7 vertically upward corresponding to the optical axis of an objective lens 11 to be described later is incident on the apex from the direction corresponding to the axis, thereby The conical mirror 12 that deflects radially and the laser light radially deflected by the conical mirror 12 are deflected vertically upward again to produce a cylindrical light beam having a constant radial width over the entire circumference. And an annular mirror 14 having a conical inner surface to be formed.

  The deflection optical system 9 includes an annular lens 15 that makes a cylindrical surface-shaped light beam incident to form a convergent light that gradually decreases the radial light beam width, and a part of the light beam in the middle of the convergence by the lens 15. Are provided with a concave mirror 16 having a conical inner surface that deflects inward in the radial direction, and a cylindrical lens 17 that converts the convergent light deflected by the concave mirror 16 into convergent light having a certain width dimension. As a result, sheet-like illumination light is formed in which the horizontal light flux width is constant and the thickness in the vertical direction is the smallest in the vicinity of the position intersecting the observation optical axis P.

  In the present embodiment, two sets of the concave mirror 16 and the cylindrical lens 17 constituting the deflection optical system 9 are arranged at positions facing each other with the sample container 2 interposed therebetween, and illumination light is irradiated from both directions with the sample X interposed therebetween. Can be done.

  The moving mechanism 10 includes a bearing 20 that supports an optical element base 18 that supports the two concave mirrors 16 and the cylindrical lens 17 of the deflecting optical system 9 so as to be rotatable around the observation optical axis P, and a base 19. , A drive gear 22 that is rotated horizontally by the motor 21, and a driven gear 23 that is fixed to the optical element base 18 and meshes with the drive gear 22.

When the drive gear 22 is rotated horizontally by driving the motor 21, the driven gear 23 that meshes with the drive gear 22 is rotated horizontally around the observation optical axis P. As a result, the optical element base 18 to which the driven gear 23 is fixed is observed. The concave mirror 16 and the cylindrical lens 17 that rotate horizontally around the optical axis P and are fixed to the optical element base 18 are rotated around the observation optical axis P.
The motor 21 is controlled by a control unit (not shown) so as to rotate the optical element base 18 around the observation optical axis P within a preset angle range in accordance with the exposure time of the camera 25 described later. Yes.

  The observation optical system 4 includes an optical axis arranged in the vertical direction at a position that coincides with the central axis of the cylindrical surface light beam constituted by the cylindrical light beam forming optical system 8, and condenses the fluorescence generated in the sample X. The objective lens 11 and a camera 25 that captures fluorescence condensed by the objective lens 11 are provided. In the figure, reference numeral 27 denotes a mirror, and reference numeral 28 denotes a filter that blocks laser light.

The operation of the sheet illumination microscope 1 according to the present embodiment configured as described above will be described below.
In order to observe a living sample X such as a zebrafish using the sheet illumination microscope 1 according to the present embodiment, the accommodating portion 6 in which the sample X to be observed by accommodating the movable stage 5 is accommodated is an observation optical axis. The sample container 2 is positioned so as to be placed on P.

  In this state, when laser light is emitted from the light source unit 7, the laser light emitted vertically upward from the light source unit 7 is incident on the conical surface of the conical mirror 12 of the cylindrical light beam forming optical system 8. The beam is uniformly deflected radially in the direction, and is deflected vertically upward by an annular mirror 14 disposed at a position surrounding the entire circumference of the conical mirror 12. As a result, a cylindrical surface light beam is formed that surrounds the outer periphery of the observation optical axis P in the entire radial direction and has a constant radial width.

  The cylindrical surface light beam is condensed by the annular lens 15 of the deflection optical system 9 arranged vertically above the annular mirror 14 and converged light whose radial width dimension gradually decreases upward. Converted. Further, only a part of the cylindrical light beam in the circumferential direction is deflected by approximately 90 ° by the concave mirror 16 and is passed through the cylindrical lens 17 so that the illumination is directed radially inward along the horizontal plane. Light is formed. The illumination light is formed in a sheet shape having a constant horizontal width dimension and gradually decreasing the vertical thickness dimension, and the illumination light deflected by the two sets of concave mirrors 16 is applied to the sample X in the sample container 2. , And incident from both directions across the sample X along the horizontal plane.

  In the incident plane through which the sheet-like illumination light passes in the sample X, fluorescence is generated by exciting the fluorescent material, and a part of the generated fluorescence emitted downward is collected by the objective lens 11. The fluorescence image is obtained by being photographed by the camera 25 after being irradiated with light and the laser light being removed by the filter 28.

  Since the illumination light has the smallest thickness in the vertical direction in the vicinity of the position intersecting the observation optical axis P, only a sufficiently thin region of the sample X can be irradiated with the laser light. The spatial resolution of the fluorescence emitted can be improved.

  According to the sheet illumination microscope 1 according to the present embodiment, the optical element base 18 is rotated in one direction over a predetermined angular range by the operation of the motor 21 within the exposure time of the camera 25. The incident angle of the sheet-like illumination light on the sample X changes in time. Thereby, there is an advantage that it is possible to suppress the occurrence of stripe-like shadows that occur when the illumination light is irradiated on the portion of the sample X where it is difficult for light to transmit due to absorption or where the light is scattered.

  Further, since the illumination light is incident from both directions with the sample X sandwiched between the two sets of the concave mirror 16 and the cylindrical lens 17, there is an advantage that it is possible to prevent the illumination from being unbalanced due to scattering and absorption in the sample X.

  When the observation of one sample X is completed, the sample container 2 is moved by the operation of the movable stage 5, the observation sample X itself or the observation position of the sample X is changed, and the observation by irradiation with the sheet-like illumination light is performed. repeat. In this case, in observations that are temporally adjacent, the sample X can be observed at high speed without waste by reversing the rotation direction of the motor 21.

  As described above, according to the sheet illumination microscope 1 according to the present embodiment, the incident angle of the illumination light can be changed without rotating the sample container 2, and the occurrence of striped shadows can be suppressed and the zebrafish can be changed. There is an advantage that it is not necessary to give a stimulus or a visual stimulus by acceleration to the living sample X. Further, since the incident angle of the illumination light is changed, an optical scanning device such as a galvanometer mirror or a polygon mirror is unnecessary, and there is an advantage that the device can be configured in a small size and at low cost.

In this embodiment, an example in which two sets of the concave mirror 16 and the cylindrical lens 17 are provided has been described, but instead of this, one set or three or more sets may be provided.
In the present embodiment, the focal position at which the sheet-like illumination light formed by each set of the deflection optical systems 9 becomes the thinnest is made to substantially coincide with the optical axis of the observation optical system 4. Thus, as shown in FIG. 3, the focal positions may be arranged at different positions. By doing in this way, it can illuminate over a wider range.

  In addition, as the moving mechanism 10, the incident angle of the illumination light on the sample X is changed by rotating the drive gear 22 meshing with the driven gear 23 fixed to the optical element base 18 by the motor 21. Alternatively, as shown in FIG. 4, the optical element base 18 may be reciprocated by the motor 21 and the crank mechanism 26 that rotate in one direction.

In the present embodiment, the conical mirror 12 and the annular mirror 14 form a cylindrical light beam over the entire circumference. However, for the light beam outside the operating range of the concave mirror 16, the surface of the conical mirror 12, etc. The light beam may not be formed by a mask (not shown) provided at an arbitrary position.
Moreover, although the thing which supports the sample container 2 so that a movement in a horizontal direction was supported was illustrated as the movable stage 5, it may replace with this and the thing supported so that a movement in a three-dimensional direction may be employ | adopted.

  Further, in the present embodiment, laser light is emitted from the light source unit 7 in a direction coinciding with the observation optical axis P, and the laser light radially deflected by the conical mirror 12 is cylindrically shaped by the annular mirror 14. Formed into a luminous flux. Instead, as shown in FIGS. 5 to 7, laser light may be incident in the horizontal direction from the outside of the observation optical axis P in the radial direction.

  In this case, the cylindrical light beam forming optical system 8 makes the laser light emitted from the exit end 30a of the optical fiber 30 that is a point light source substantially parallel light, and the collimating lens 31 makes the light substantially parallel light. The first cylindrical lens 32 having a negative power that once spreads the converted laser beam in the horizontal direction, and the convergent light that condenses the laser beam spread by the first cylindrical lens 32 on the observation optical axis P. A second cylindrical lens 33 having a positive power to be converted into a second cylindrical lens 33 and a partial cylinder heading vertically upward by deflecting the laser light from the second cylindrical lens 33 by 90 ° by the outer surface of the cone centered on the observation optical axis P What is necessary is just to provide the annular mirror 14 which forms a planar light beam.

  Thereby, using the deflection optical system 9 similar to that in FIG. 1, as shown in FIG. 7, the sheet-like illumination light having a constant width and having a focal point near the observation optical axis P as in FIG. Can be formed. The moving mechanism 10 can change the incident angle of the illumination light by moving the concave mirror 16 and the cylindrical lens 17 within the range of the partial cylindrical light beam. The annular mirror 14 and the annular lens 15 may be formed in an arc shape in a range necessary for forming a cylindrical surface light beam.

  Further, in FIG. 5, a cylindrical surface-shaped light beam is formed by the annular mirror 14 and the light beam is converged by the annular lens 15, but instead, as shown in FIG. An annular or arcuate mirror 34 having a reflecting surface composed of a parabolic concave surface in which the functions of the annular mirror 14 and the annular lens 15 are integrated may be employed. Thereby, the number of parts can be reduced.

  Further, as shown in FIG. 9, the functions of the annular lens 15 and the concave mirror 16 are integrated, and the vicinity of the observation optical axis P is obtained from a cylindrical surface-shaped light beam having a constant thickness formed by the annular mirror 14. Alternatively, a parabolic mirror 35 that forms a sheet-like illumination light that is focused on may be employed. This also reduces the number of components, and can form illumination light with a larger emission NA, and provide a sheet illumination microscope with high spatial resolution by making the illumination light at the focal position thin and good. There is an advantage that can be.

In the present embodiment, the central axis of the cylindrical light beam is made to coincide with the single observation optical axis P. However, instead of this, as shown in FIGS. A plurality of systems 4 may be provided.
In the example shown in FIGS. 10 and 11, the four objective lenses 11 having the observation optical axis P parallel to the central axis of the cylindrical light beam are arranged at equal intervals in the circumferential direction. In the figure, reference numeral 36 denotes a cylindrical light shielding plate.

  By doing in this way, the cylindrical sample container 2 centering on the central axis of a cylindrical surface light beam is employable as the sample container 2. The sample container 2 is provided with eight storage sections 6 that are partitioned in the circumferential direction, and is 45 ° by the movable stage 5 in order to observe the sample X stored in the storage section 6 adjacent in the circumferential direction. If the sample container 2 is moved only once, all the samples X in the eight accommodating portions 6 can be observed.

  Also, as shown in FIGS. 12 and 13, two sets of observation optical systems 4 may be provided. In this way, two samples X are observed simultaneously using a multi-well plate (sample container) 37 having eight wells (accommodating portions) 6 in 4 rows and 2 columns, and the multi-well is moved by the movable stage 5. By translating the plate 37 in the direction along the row of the wells 6, the samples X in all the wells 6 can be observed. By making the interval of the objective lens 11 coincide with the well interval of a normal multiwell plate, there is an advantage that an existing dispensing device or the like can be used.

  Further, in this embodiment, the optical element base 18 to which the concave mirror 16 and the cylindrical lens 17 are attached is rotated around the observation optical axis P. Instead, this is shown in FIGS. In this manner, the observation optical axis P may be sandwiched between the cylindrical lens 17 and the center lens disposed on the opposite side. Even in this case, the focal position of the sheet-like illumination light is preferably arranged in the vicinity of the position intersecting the observation optical axis P.

  By doing so, the incident angle of the sheet-like illumination light can be changed so as to cross the observation optical axis P, and even when observing a larger sample X, the movable stage 5 is not operated. The entire sample X can be irradiated with illumination light. It can be configured more simply and at a lower cost than a movable stage using a linear motion mechanism such as an XY stage.

  In the example shown in FIGS. 14 and 15, the case where two sets of cylindrical light beam forming optical system 8 and deflecting optical system 9 are provided has been described. Instead of this, the present invention may be applied to the case where one set or three or more sets of cylindrical light beam forming optical system 8 and deflection optical system 9 are provided. By using two or more sets of cylindrical light beam forming optical system 8 and deflecting optical system 9, the irradiation amount of illumination light per unit time to sample X caused by the difference in moving speed of illumination light due to the difference in distance from the central axis Can be reduced.

  Also, as shown in FIGS. 16 and 17, the deflection optical system 9 includes two or more concave mirrors (deflection members) 16, a conical surface mirror (deflection member) 29, and a parabolic mirror (deflection members, optical elements). 35 may be arranged in series. In the example shown in FIG. 16, three mirrors 16, 29, and 35 are arranged in series. Here, the concave mirror 16 has negative power, the conical mirror 29 has positive power, and the parabolic mirror 35 has positive power.

  In this way, the diameter of the cylindrical surface light beam formed by the annular mirror 14 is made relatively large in order to avoid the correction ring adjustment unit 38 and the like disposed in the vicinity of the objective lens 11. Even in the case where it is unavoidable, the outer diameter of the upper end where the sheet-like illumination light is emitted can be reduced. Thereby, as shown in FIGS. 16 and 17, it is possible to observe the sample X accommodated in the sample container 2 in which the interval between the accommodating portions 6 is narrow.

  In the example shown in FIG. 16, if the three mirrors 16, 29, and 35 are fixed to the optical element base 18 as a module and fixed to the inner ring of the bearing 20 in a replaceable manner with screws or the like, it is shown in FIG. As described above, the spatial resolution can be switched by switching to another module that emits illumination light having a different focal length (exit NA).

  Further, as shown in FIGS. 19 and 20, a solution chamber 39 for immersing the sample container 2 in an optically transparent solution may be provided, and the optical element base 18 may be arranged in the solution in the solution chamber 39. . Then, an immersion objective lens 40 may be employed as the objective lens, and the bottom surface of the solution chamber 39 may be configured with an optically transparent material. The liquid in the solution chamber 39, the solution in the sample container 2, and the liquid interposed between the immersion objective lens 40 and the bottom surface of the solution chamber 39 are used as liquids having substantially the same refractive index. Even if the incident angle of the illumination light is changed by the operation, there is an advantage that a good image can be acquired while suppressing the occurrence of aberration.

  In this case, in addition to rotating the entire solution chamber 39, as shown in FIG. 21, only a part of the bottom surface of the solution chamber 39 is rotatably provided by the liquid shield bearing 47, and the mirror of the deflection optical system 9 is provided. 16 and 42 may be configured such that the light beam passes through the bottom surface of the solution chamber 39. By doing so, it is not necessary to move the entire solution with a large mass as compared with the case where the entire solution chamber 39 is rotated, and it is possible to perform stable observation by preventing generation of waves due to the movement. There is an advantage that you can.

DESCRIPTION OF SYMBOLS 1 Sheet illumination microscope 2 Sample container 3 Illumination optical system 4 Observation optical system 5 Movable stage 8 Cylindrical light beam formation optical system 9 Deflection optical system 10 Movement mechanism 16 Concave mirror (module, deflection member)
29 Conical mirror (deflection member)
33 Second cylindrical lens (optical element)
35 Parabolic mirror (module, deflection member, optical element)
P Observation optical axis X Sample

Claims (12)

An illumination optical system that makes sheet-like illumination light incident on the sample contained in the sample container;
An observation optical axis perpendicular to an incident plane of the illumination light incident by the illumination optical system, and an observation optical system that forms an image of the sample by light generated in the sample by irradiation of the illumination light,
The illumination optical system forms a cylindrical light beam forming optical system that forms a cylindrical surface light beam having a central axis parallel to the observation optical axis, and a part of the light beam formed by the cylindrical light beam forming optical system is the incident plane. A sheet illumination microscope comprising: a deflection optical system that deflects in a direction along the axis to form the illumination light; and a moving mechanism that moves the deflection optical system around the central axis.   The sheet illumination microscope according to claim 1, wherein the cylindrical light beam forming optical system forms a cylindrical light beam having a central axis substantially coinciding with the observation optical axis.   The sheet illumination microscope according to claim 1, wherein the cylindrical light beam forming optical system forms the cylindrical surface light beam on the opposite side of the central axis across the observation optical axis.   The sheet illumination microscope according to claim 3, wherein the illumination optical system includes a plurality of sets of the deflection optical system and the moving mechanism with respect to a single cylindrical light beam forming optical system.   The sheet illumination microscope according to claim 1, further comprising a movable stage that moves the sample container in a direction orthogonal to at least the observation optical axis.   The sheet illumination microscope according to claim 5, wherein the sample container can accommodate the sample at different positions along a moving direction of the movable stage.   The sheet illumination microscope according to any one of claims 1 to 6, wherein the deflection optical system forms the illumination light including a light beam having a certain width in the incident plane.   The sheet illumination according to any one of claims 1 to 7, wherein the deflection optical system forms a light beam that converges in a plane including an illumination optical axis of the illumination light and the observation optical axis along the incident plane. microscope. A plurality of the deflection optical systems;
The sheet illumination microscope according to claim 8, wherein each of the deflection optical systems forms the illumination light condensed at different positions.   10. The deflecting optical system includes an optical element having a positive power for converging the illumination light, and is configured by a replaceable module for switching a focal length of the illumination light. The sheet illumination microscope described.   The sheet illumination microscope according to any one of claims 1 to 10, comprising a plurality of the observation optical systems.   The sheet illumination microscope according to claim 1, wherein the deflection optical system includes two or more deflection members arranged in series.

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