Light sheet microscope with interchangeable optical elements

Abstract

To provide a light sheet microscope having an optical system including a first optical element, a second optical element, and a mirror assembly, especially a slope microscope or a SCAPE microscope, the microscope solving an issue of the solution of an existing technique that the existing technique cannot response to different measurement conditions and needs to be adapted inefficiently.SOLUTION: The solution of an existing technique is improved by provision of a first operational state. In the first operational state, an optical system is formed to transmit illumination light to a first sample volume body through a first optical element along a first illumination optical path and also is formed to transmit observation light to a detector from the first sample volume body along a first observation optical path. In the second operational state, the optical system is formed to transmit illumination light to a second sample volume body through a second optical element along a second illumination optical path and also is formed to transmit observation light to a detector from the second sample volume body along a second observation optical path.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a light sheet microscope, in particular an oblique plane microscope or SCAPE microscope, having an optical system with at least one first optical element, at least one second optical element and a mirror assembly. [Background technology]

[0002] Several light sheet microscopes are known in the art, where the parameters of the microscope are generally fixed, such as magnification, numerical aperture, etc. Changing such parameters of a light sheet microscope is inefficient and time-consuming. Summary of the Invention [Problem to be solved by the invention]

[0003] It is therefore an aspect of the present invention to provide a light sheet microscope that facilitates modification of microscope parameters, increasing the versatility of the microscope. [Means for solving the problem]

[0004] The above problem with the light sheet microscope mentioned at the beginning is solved by the present invention in that in a first operating state of the microscope, the optical system is configured to transmit illumination light along a first illumination light path through at least one first optical element to a first sample volume and to transmit observation light along a first observation light path from the first sample volume to a detection device, and in a second operating state of the microscope, the optical system is configured to transmit illumination light along a second illumination light path via a mirror assembly through at least one second optical element to a second sample volume and to transmit observation light along a second observation light path from the second sample volume to a detection device.

[0005] The light sheet microscope according to the present invention therefore has the advantage that it makes it possible to select at least an optical element from a first optical element and a second optical element, through which a sample volume is illuminated, and through which observation light from the sample volume is received and transmitted towards a detection device.

[0006] The light sheet microscope according to the invention can be further improved by a number of additional technical features, which are described below, which can be combined with one another in any way or can be omitted if the technical effects obtained by the omitted technical features are not relevant to the present invention.

[0007] The light sheet microscope may include three or more optical elements, and the present invention allows one of these optical elements to be selected. Thus, the light sheet microscope allows one optical element to be selected from any number of optical elements. Selecting one of these optical elements is understood to mean providing an operating state of the microscope in which the illumination light path and the observation light path pass through this optical element.

[0008] The optical elements may be objective lenses. The optical elements may differ from each other in at least one of the directions of the corresponding optical axes, the magnification, the numerical aperture, and the material and corresponding optical properties of the optical elements. This list of properties is not exhaustive, and for at least two optical elements used in the light sheet microscope, other properties of the optical elements may differ.

[0009] A light sheet microscope being in a first operating state or in a second operating state is understood to mean that one or several features of the light sheet microscope are in a first position or in a second position.

[0010] In particular, the mirror assembly is positionable in a first operational position in a first operational state and movable to a second operational position in a second operational state.

[0011] Thus, the light sheet microscope according to the invention is switchable between a first illumination path and a second illumination path of at least two different illumination paths. Any number of different paths is possible, the number of which may be limited by the geometry of the microscope and the available space.

[0012] The at least two different illumination light paths may be at least partially collinear, i.e., oriented identically to one another. Preferably, the light paths may be identical from the illumination source to the second operational position of the mirror assembly.

[0013] Each observation light path can be defined by a corresponding illumination light path. The first illumination light path, through which illumination light is transmitted to the first sample volume, can thus define the first observation light path or at least define the positional limit of the first observation light path. Preferably, in the sample volume, the observation light path can be oriented substantially perpendicular to the illumination light path. More preferably, the angle between the observation light path and the illumination light path is between 0° and 90°. Even more preferably, this angle is between 45° and 90°, for example about 60°.

[0014] Furthermore, the first illumination light path and the first observation light path are assigned to the first sample volume. Correspondingly, the second illumination light path and the second observation light path are assigned to the second sample volume. Correspondingly, a third sample volume or a fourth sample volume is provided with a corresponding light path. The number of possible sample volumes is limited only by the geometrical limitations of the light sheet microscope.

[0015] In the first operational position of the mirror assembly, the first illumination light path can bypass the mirror assembly.

[0016] In one embodiment of the light sheet microscope according to the present invention, the mirror assembly may be configured to be movable at least one of linearly and rotationally from a first operating position to a second operating position.

[0017] Thus, in a first operating state, the first illumination light path and the first observation light path may bypass the mirror assembly, whereas in a second operating state, the mirror assembly may be positioned in the second illumination light path and the second observation light path.

[0018] As mentioned above, the two or more illumination light paths and the two or more observation light paths may overlap, and the second operational position of the mirror assembly may define a branch point for the illumination light paths or a joining point for the observation light paths. Preferably, only one illumination light path and only one observation light path is selected by the operational position of the mirror assembly depending on the operational state of the light sheet microscope.

[0019] The mirror assembly may comprise at least one mirror. It is also conceivable that the mirror assembly comprises two or more mirrors for redirecting the illumination light path and the corresponding observation light path to the corresponding optical element. Apart from the first illumination light path and the first observation light path (both bypassing the mirror assembly), the mirror assembly may further define at least one second illumination light path and a second observation light path in a second operating position. The mirror assembly of the light sheet microscope may further define a third illumination light path and a third observation light path in a third operating position. Correspondingly, fourth, fifth, up to nth operating positions of the mirror assembly may be possible. In each operating position, a different mirror (or a different number of mirrors) may be provided in the illumination light path and the observation light path.

[0020] The mirror assembly may be in a pre-adjusted state for each operating position. If the light sheet microscope according to the invention has more than one operating position, for example three operating positions, the first mirror of the mirror assembly may be arranged in the illumination light path and the observation light path in the second operating position of the mirror assembly, whereas the second mirror of the mirror assembly may be arranged in the illumination light path and the observation light path in the third operating position of the mirror assembly. Thus, in the third operating position of the mirror assembly, the first mirror of the mirror assembly may be bypassed by the third illumination light path and the third observation light path. As a result, different mirrors, each capable of realizing a pre-adjustment, can be used to direct the illumination light and the observation light along the second, third, up to n-th illumination light path and the observation light path.

[0021] Preferably, the back focal plane / pupil of the first and second optical elements (and of other optical elements, such as the third, fourth, up to nth optical elements in other embodiments of the microscope) are arranged at the same position of the light sheet microscope. In general, the back focal plane of an optical element is the Fourier plane of the focal plane of the optical element, which is in the sample, or in another plane that is conjugate with this plane. Thus, for example for the first optical element, the corresponding back focal plane is the focal plane located on the opposite side to the first sample volume. This definition of the back focal plane is applicable to the second optical element and the second sample volume, and also to other possible, for example third, etc. optical elements and sample volumes. It is further particularly advantageous if the back focal planes of all optical elements, i.e. the first, second, third, etc., nth optical elements, coincide with another focal plane of the objective lens that generates the oblique virtual image, which is then imaged to the detection device by an optical assembly that is obliquely oriented with respect to the other optical components.

[0022] The focal angles of the optical elements (first, second, third, etc.) may be the same, i.e. M_1 / n_1=M_2 / n_2 applies, where M is the magnification given by the ratio of the tube lens to the objective lens M=f_tube / f_objective. So as an example: f_tube=200mm, f_objective=10mm → M=20, the variable n is the refractive index for which the lens is optimized, i.e. n_1=1.33 for a water immersion lens. If the above formula does not hold, an intermediate telescope may be required. According to this disclosure, the proposed way to make this formula hold is to have a matching focal length f_tube of the tube lens in each beam path.

[0023] The optical elements are preferably configured or adjusted such that their positions are fixed and their focal planes / pupils overlap as described above. Thus, the operational state of the light sheet microscope can be easily changed by a guided linear movement of the optical assembly without the need for realignment. Prior art solutions, for example using a rotating revolver, cannot provide such precision and require inefficient and time-consuming realignments.

[0024] In another embodiment of the light sheet microscope, the mirror assembly is configurable to be moved into the first illumination light path along an insertion direction, the insertion direction comprising: substantially perpendicular to the first illumination light path; substantially parallel to the reflective surface of the mirror assembly, and in the second operating state, substantially perpendicular to the reflective surface of the mirror assembly.

[0025] The mirror assembly is preferably linearly movable into the first illumination optical path.

[0026] In another embodiment of the light sheet microscope, the optical system may have a first tube lens and a second tube lens, the first tube lens may be arranged in the first illumination light path, and the second tube lens may be arranged in the second illumination light path. Correspondingly, a third tube lens or a fourth tube lens may be arranged in the third illumination light path or the fourth illumination light path, respectively. Preferably, in each operating state of the light sheet microscope, one tube lens is utilized, through which the illumination light path (preferably also the observation light path) is directed.

[0027] The tube lens may be adapted to the corresponding first, second, third or nth optical element in terms of numerical aperture, focal length or magnification. The mirror assembly and the first, second, third or nth tube lens in the first, second, third or nth operating position may be pre-adjusted. Such pre-adjustment may preferably result in a partially coaxial arrangement of all illumination light paths from the light source to the branching point (at which point at least the first mirror of the mirror assembly is located in the second, third or nth operating position) and a partially coaxial arrangement of all observation light paths from the coupling point to the detection device.

[0028] Most preferably, the optical elements and / or corresponding tube lenses provided in one of the illumination light paths are mounted in a fixed position relative to the microscope, with only the mirror assembly being moved.

[0029] However, at least one of the second tube lens and the at least one second optical element can be configured to be moved with the mirror assembly from a first operating position to a second operating position, in which the second tube lens and / or the at least one second optical element is inserted into the second illumination light path. The mirror assembly can include another tube lens (e.g., a third tube lens). One or more of the other tube lenses can be configured to move with the mirror assembly.

[0030] It is also conceivable that the mirror assembly comprises different independently movable mirrors. With each of the independently movable mirrors mentioned above, a corresponding independently movable tube lens may be provided.

[0031] Exemplarily, the first tube lens can be provided in the first illumination light path (preferably also in the first observation light path), i.e. when the mirror assembly is bypassed. In a second operating position of the mirror assembly, the first mirror (optionally together with the second tube lens) can be moved into the first illumination light path, thereby defining a second illumination light path. A third operating position of the mirror assembly is conceivable, in which the second mirror (optionally together with the third tube lens) can be moved into the first illumination light path, thereby defining a third illumination light path. The movement of the first mirror (and the second tube lens) can be independent of the movement of the second mirror (and the third tube lens). This movement can in particular be performed along different directions. These directions can be oriented perpendicular to one another.

[0032] In another embodiment of the light sheet microscope, in the first illumination path, a first tube lens of the optical system may be arranged between the at least one first optical element and the second operating position of the mirror assembly, such that the first tube lens can be bypassed when the mirror assembly is moved to the second operating position.

[0033] Thus, when a light sheet microscope according to the invention is considered, having a first operating state and a second operating state (without excluding another operating state), different possibilities for switching between states are conceivable. Firstly, only the mirror assembly is moved from the first operating position to the second operating position to transmit the illumination light along the second illumination light path, which transmits the illumination light to the second sample volume through at least one second optical element by the mirror assembly. Here, the second optical element (which may be an objective lens) and the second tube lens are fixed relative to the light sheet microscope and are not moved when changing the operating state. Secondly, the second tube lens is movable together with the mirror assembly, while the second optical element remains fixed. Thirdly, the mirror assembly, the tube lens and the second optical element are moved together to the second operating position.

[0034] Therefore, a light sheet microscope can have a combination of tube lenses and optical elements for each operating state.

[0035] Each of these combinations is configured for observation of a sample volume, which may differ in at least one of a position and an orientation for each operating state.

[0036] For example, it is conceivable that the light sheet microscope according to the invention has three operating states, where in a first operating state the light sheet microscope is configured for viewing from the side (i.e. perpendicular to the up-down direction defined by gravity), whereas in a second operating state the sample provided in the second sample volume can be viewed from above, i.e. with illumination along the direction of gravity. Additionally, in a third operating state the light sheet microscope can be configured for viewing the sample from below, i.e. with illumination against the direction of gravity.

[0037] In another embodiment of the light sheet microscope according to the invention, the ratio of the focal length of the second optical element to the focal length of the second tube lens is substantially equal to the ratio of the focal length of the at least one first optical element to the focal length of the first tube lens, thereby obtaining equal optical path lengths of the illumination and observation optical paths for two operating states (preferably, more generally, for all operating states).

[0038] Additionally, the mirror assembly may include at least one adjustable mirror configured to be adjustable in at least one of an angle relative to the second illumination optical path and a position along the second illumination optical path, thereby facilitating any readjustment required after a change in the operational state of the light sheet microscope from a first operational state to a second operational state.

[0039] The optical system of the light sheet microscope according to the invention may comprise a pair of adjustable mirrors, each of which is configured to adjust at least one of the angle with respect to the first and second illumination light paths and the position along the first and second illumination light paths. By using such an adjustable mirror pair, not only the position of the light path in the optical element or tube lens can be adjusted, but also the angle of the light path through this element.

[0040] In another advantageous embodiment of the light sheet microscope, the first optical element may be a first objective lens, the second optical element may be a second objective lens, and the first objective lens and the second objective lens are Aperture, Working distance, Pupil diameter, and ·Focal length At least one of them is different.

[0041] Furthermore, the optical axis of the first optical element and the optical axis of the second optical element are Substantially parallel to each other substantially antiparallel to each other, and Substantially perpendicular to each other The axial orientation of the axial direction of the gyro can be one of the following:

[0042] As explained above, the direction in which the sample volume is observed may be different for different operating states of the light sheet microscope. In the parallel orientation, the observation is performed in the same direction for two (or all) operating states of the light sheet microscope, whereas in the antiparallel orientation, the direction of observation is opposite, e.g. downward and upward for the first and second operating states, respectively. The third case above is the observation of the sample volume from the side, while in another operating state the observation is performed upward or downward.

[0043] In another advantageous embodiment of the light sheet microscope according to the present invention, the optical system may have at least two scanning mirrors configured to be adjustable in at least one of angle and position relative to the first and second illumination light paths, which scanning mirrors are configured to scan the first and second sample volumes with the light sheet generated in the sample volume.

[0044] Although some aspects have been described in the context of an apparatus, it will be apparent that these aspects also represent a description of a corresponding method, where a block or apparatus corresponds to a step or feature of a step. Similarly, aspects described in the context of a step also represent a description of a corresponding block or item or feature of a corresponding apparatus. Some or all of the steps may be performed by (or using) a hardware apparatus, such as, for example, a processor, microprocessor, programmable computer, or electronic circuitry. In some embodiments, any one or more of the crucial steps may be performed by such an apparatus.

[0045] Depending on certain implementation requirements, the embodiments of the present invention can be implemented in hardware or software. The implementation can be performed by a non-transitory recording medium, such as a digital recording medium, for example a floppy disk, a DVD, a Blu-ray, a CD, a ROM, a PROM and EPROM, an EEPROM or a FLASH memory, on which electronically readable control signals are stored, which cooperate (or can cooperate) with a programmable computer system to implement the respective methods. Thus, the digital storage medium can be computer readable.

[0046] Some embodiments of the present invention include a data carrier having electronically readable control signals capable of cooperating with a programmable computer system to perform any of the methods described herein.

[0047] Generally, embodiments of the present invention can be implemented as a computer program product comprising program code which is operable to perform any of the methods when the computer program product is run on a computer, the program code may for example be stored on a machine readable carrier.

[0048] Another embodiment comprises the computer program for performing any of the methods described herein, stored on a machine readable carrier.

[0049] In other words, an embodiment of the present invention is, therefore, a computer program having a program code for performing any of the methods described herein, when the computer program runs on a computer.

[0050] Therefore, another embodiment of the present invention is a recording medium (or data carrier or computer readable medium) containing a computer program stored thereon for performing any of the methods described herein when executed by a processor. The data carrier, digital recording medium or recording medium is typically tangible and / or non-transitory. Another embodiment of the present invention is an apparatus as described herein, including a processor and a recording medium.

[0051] A further embodiment of the invention is therefore also a data stream or a sequence of signals representing the computer program for performing any of the methods described herein, the data stream or the sequence of signals being for example adapted to be transmitted via a data communication connection, for example the Internet.

[0052] Another embodiment comprises a processing means, for example a computer, or a programmable logic device configured to or adapted to perform any of the methods described herein.

[0053] Another embodiment comprises a computer having the computer program installed thereon for performing any of the methods described herein.

[0054] Another embodiment of the invention includes an apparatus or system configured to transfer (e.g., electronically or optically) a computer program for implementing any of the methods described herein to a receiver. The receiver may be, for example, a computer, a mobile device, a storage device, etc. The apparatus or system may, for example, include a file server to transfer the computer program to the receiver.

[0055] In some embodiments, a programmable logic device (e.g., a field programmable gate array) may be used to perform some or all of the functionality of the methods described herein. In some embodiments, a field programmable gate array may cooperate with a microprocessor to perform any of the methods described herein. In general, the methods are advantageously performed by any hardware apparatus.

[0056] The light sheet microscope according to the invention will be described below with reference to the figures attached in the drawings, in which the same technical features and features having the same technical effect are provided with the same reference signs. The technical features of the different figures can be combined or omitted in any way. The figures show exemplary embodiments of the invention, which do not limit the scope of patent protection defined by the claims. [Brief description of the drawings]

[0057] [Figure 1] FIG. 1 shows a first exemplary embodiment of an oblique microscope according to the present invention. [Diagram 2] FIG. 2 shows a second exemplary embodiment of an oblique microscope according to the present invention. [Diagram 3] FIG. 1 shows a first exemplary embodiment of a SCAPE microscope according to the invention. [Figure 4] FIG. 2 shows a second exemplary embodiment of a SCAPE microscope according to the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0058] In the following description of the figures, each technical feature is numbered with a leading number indicating in which figure this technical feature is shown. However, this does not exclude, by way of example, the presence of a feature having, for example, reference number 405 in figures 1 to 3. This reference number 405 does not exclude the presence of this feature in the preceding figures 5, 6, etc. Thus, the same technical feature present in more than one figure will be referred to by reference numbers 105, 205, 305, 405, ..., x05, with the "x" indicating the overall number of the figure.

[0059] FIG. 1 shows a light sheet microscope 101, in particular an oblique microscope 102, having an optical system 104 with at least one first optical element 105, at least one second optical element 113, and a mirror assembly 114.

[0060] FIG. 1 shows two operating states of the oblique microscope 102, namely a first operating state 106, in which the optical system 104 is configured to transmit illumination light 107 from a light source 107a along a first illumination light path 108, through a first optical element 105, and to a first sample volume 109.

[0061] In the first sample volume 109 a light sheet 128 is generated which illuminates a planar portion within the first sample volume 109 .

[0062] Also shown in Fig. 1 is a coordinate system showing the directions y and z. In a first operating state 106, the first sample volume 109 is illuminated along the z direction, which may correspond to illumination from the side, which can be used, for example, if it is desired to examine root growth.

[0063] Within the first sample volume 109, observation light 110 is generated and transmitted via a first observation light path 111 through the first optical element 105, which generates an inclined virtual image 128a, which is then imaged correctly (i.e., flat and undistorted) onto the detection device 112 by the optical detection assembly 112a.

[0064] In the first operational state 106, the mirror assembly 114 is in a first operational position 116. In the first operational position 116, the mirror assembly 114 is bypassed by the first illumination light path 108, i.e., the mirror assembly 114 may be located, for example, in the bypass position 114b shown in Figure 1. Different bypass positions 114b are contemplated, for example, in the plane of the drawing or out of the plane of the drawing.

[0065] The mirror assembly 114 is movable along an insertion direction 120 to a second operating position 117. This can be accomplished by a translation device 118, such as a linear translation stage 119.

[0066] The insertion direction 120 can be oriented parallel to the reflective surface 114a. The insertion direction 120 can also be oriented perpendicular to the first illumination light path 108 or substantially perpendicular to the reflective surface 121 of the mirror assembly 114 in the second operating state 115.

[0067] In a second operating state 115 of the microscope 101, the optical system 104 is configured to transmit the illumination light 107 along a second illumination optical path 108a via the mirror assembly 114. The illumination light 107 is transmitted through at least one second optical element 113 to a second sample volume 109a. Similar to the first sample volume 109, a light sheet 128 is generated and the observation light 110 is transmitted from the second sample volume 109a to the detection device 112 along a second observation optical path 111a.

[0068] 1, in the first portion 129 of the microscope 101, the first illumination light path 108 at least partially corresponds to, overlaps with and is collinear with the second illumination light path 108a. Similarly, in the first portion 129, i.e., the first observation light path 111 and the second observation light path 111a are collinear with each other.

[0069] FIG. 2 shows a second embodiment of an oblique microscope 202 similar to that shown in FIG.

[0070] 2 further comprises a first tube lens 222 and a second tube lens 223. The first tube lens 222 is provided in both the first illumination optical path 208 and the first observation optical path 211.

[0071] The second tube lens 223 is used (ie, light is transmitted through it) in the second operational state 215 , ie, when the mirror assembly 214 is moved to the second operational position 217 .

[0072] It should be noted that the refraction of the light path is shown diagrammatically. Moreover, the operating state referred to as the first operating state 206 may correspond to a second operating state 215 in another embodiment of the light sheet microscope 201 according to the invention. Thus, in this other embodiment (not shown) of the light sheet microscope 201, a mirror assembly may be provided in the first observation light path 211 and in the first illumination light path 208 in the first operating state 206, whereas the mirror assembly 214 may be bypassed in the second operating state 215.

[0073] In FIG. 3 a light sheet microscope 301 according to the invention in the form of a SCAPE microscope 303 is shown.

[0074] The SCAPE microscope 303 has a light source 307a that provides illumination light 307 along a first illumination path 308. Again, in a first portion 329, the first illumination path 308 is collinear with a second illumination path 308a.

[0075] The illumination light 307 is transmitted via another tube lens 330 to a pair 331 of adjusting mirrors 332, which may simultaneously be scanning mirrors 333 for scanning the first sample volume 309 or the second sample volume 309a.

[0076] In the embodiment shown in FIG. 3, an alternative second tube lens 330a is shown.

[0077] The mirror assembly 314 is shown in a first operational position 316 and a second operational position 317. In the first operational state 306 the first sample volume 309 is viewed from below, whereas in the second operational state 315 the second sample volume 309a is viewed from above.

[0078] For each operating state 306, 315 a different optical element is provided, namely a first optical element 305 and a second optical element 313.

[0079] The first operative position 316 and the second operative position 317 of the mirror assembly 314 are depicted overlapping, with the mirror assembly 314 in the first operative position 316 showing the first mirror 334 and the mirror assembly 314 in the second operative position 317 showing the second mirror 335. Thus, Figure 3 does not show the same mirror in different positions 316, 317.

[0080] In this embodiment, either the first mirror 334 or the second mirror 335 of the mirror assembly 314 can be inserted into the optical path by moving the mirror assembly 314 along an insertion direction 320 that lies in the plane of the drawing.

[0081] The generation of the light sheet 328 and the collection of the observation light 310, which in this case is transmitted either via the first observation path 311 or the second observation path 311a, is performed similarly to the previous figures. In the first portion 329, the first illumination path 308 and the second illumination path 308a, as well as the first observation path 311 and the second observation path 311a, are collinear with each other.

[0082] The optical detection assembly 312a images the oblique virtual image 328a onto the detection device 312, where the SCAPE microscope 303 further includes another objective lens 336 that images the oblique virtual image 328a.

[0083] 4 also shows a SCAPE microscope 403, where there is no separate second tube lens 330a as in FIG. 3, but rather a telescope 437. Furthermore, in the SCAPE microscope 403 of FIG. 4, a first tube lens 422 and a second tube lens 423 are provided between the first optical element 405 and the mirror assembly 414 or between the second optical element 413 and the mirror assembly 414.

[0084] In this embodiment, the mirror assembly 414 is also moved in the illumination light path 408, 408a along the insertion direction 420. A first tube lens 422 and a second tube lens are movable along the insertion direction 420 with the mirror assembly 414.

[0085] The mirror assembly 414 again has a first mirror 434 disposed in the beam path in the first operational state 406 and a second mirror 435 disposed in the beam path in the second operational state 415 .

[0086] It is further conceivable to combine the microscopes 101, 201, 301, 401 and in particular their positions of the first sample volume 109-409 and the second sample volume 109a-409a. Thus, the SCAPE microscope 403 of Fig. 4 allows in another embodiment to provide three operating states and therefore three operating positions of the mirror assembly 414, and to provide three different sample volumes, for example the first sample volume 409, the second sample volume 409a and the third sample volume 409b. The third sample volume 409b is shown in Fig. 4 with dashed lines.

[0087] The light sheet microscope 101-401 thus allows switching between different optical elements 105-405 and 113-413 (a third optical element, not shown, may be required for observation in a third sample volume 409b) in order to observe one sample volume with different parameters and / or from different directions. When the operating state of the light sheet microscope 101-401 is changed, different illumination and observation optical paths are selected. Within these different optical paths, optical components such as optical elements and tube lenses are pre-adjusted, so that no readjustment is necessary after the operating state is changed. [Explanation of symbols]

[0088] 101 Light Sheet Microscopy 102 Oblique microscope 103 SCAPE Microscope 104 Optical system 105 First Optical Element 106 First operating state 107 Illumination 107a light source 108 First illumination light path 108a Second illumination light path 109 First sample volume 109a Second sample volume 110 Observation Light 111 First observation path 111a Second observation optical path 112 Detection Device 112a Optical detection assembly 113 Second Optical Element 114 Mirror Assembly 114a reflective surface 114b Bypass position 115 Second operating state 116 First operating position 117 Second operating position 118 Translation device 119 Linear translation stage 120 Insertion direction 121 Reflection surface 1 24 First Objective Lens 125 Second Objective Lens 126 Optical axis of the first optical element 127 Optical axis of second optical element 128 Light Sheet 128a Virtual image of slope 129 First Part 222 First Tube Lens 223 Second Tube Lens 330 Another tube lens 330a Another second tube lens 331 vs. 332 Adjustable Mirror 333 Scan Mirror 334 First Mirror 335 Second Mirror 336 Another Objective Lens 409b Third sample volume

Claims

1. A light sheet microscope (101) having an optical system (104) with at least one first optical element (105), at least one second optical element (113), and a mirror assembly (114), in a first operating state (106) of the microscope (101), the optical system (104) is configured to transmit illumination light (107) along a first illumination optical path (108) through the at least one first optical element (105) to a first sample volume (109) and to transmit observation light (110) along a first observation optical path (111) from the first sample volume (109) to a detection device (112), in a second operating state (115) of the microscope (101), the optical system (104) is configured to transmit the illumination light (107) along a second illumination optical path (108a), via the mirror assembly (114), through the at least one second optical element (113) to a second sample volume (109a) different from the first sample volume (109), and to transmit observation light (110) along a second observation optical path (111a) from the second sample volume (109a) to the detection device (112); Light sheet microscope (101).

2. the mirror assembly (114) is located in a first operating position (116) in the first operating state (106) and is moved to a second operating position (117) in the second operating state (115); The light sheet microscope (101) according to claim 1.

3. In the first operating position (116), the first illumination optical path (108) bypasses the mirror assembly (114). The light sheet microscope (101) according to claim 2.

4. the mirror assembly (114) is configured to be movable at least one of linearly and rotationally from the first operating position (116) to the second operating position (117); A light sheet microscope (101) according to claim 2 or 3.

5. In the first operating state (106), the first illumination optical path (108) and the first observation optical path (111) bypass the mirror assembly (114), and in the second operating state (115), the mirror assembly (114) is disposed in the second illumination optical path (108a) and the second observation optical path (111a). A light sheet microscope (101) according to any one of claims 2 to 4.

6. The mirror assembly (114) is configured to be moved into the first illumination light path (108) along an insertion direction (120), the insertion direction (12) being: - substantially perpendicular to the first illumination light path (108); - substantially parallel to the reflective surface (114a) of the mirror assembly (114); and in the second operating state (115), substantially perpendicular to a reflective surface (121) of the mirror assembly (114); One of the A light sheet microscope (101) according to any one of claims 2 to 5.

7. The optical system (104) includes a first tube lens (222) and a second tube lens (223); the first tube lens (222) is provided in the first illumination optical path (108), and the second tube lens (223) is provided in the second illumination optical path (108a); A light sheet microscope (101) according to any one of claims 1 to 6.

8. The optical system (104) has a first tube lens (222) and a second tube lens (223), the first tube lens (222) is provided in the first illumination optical path (108), and the second tube lens (223) is provided in the second illumination optical path (108a); at least one of the second tube lens (223) and the at least one second optical element (113) is configured to move with the mirror assembly (114) from the first operating position (116) to the second operating position (117); In the second operating position (117), the second tube lens (223) and / or the at least one second optical element (113) are inserted into the second illumination light path (108a). A light sheet microscope (101) according to any one of claims 2 to 6.

9. In the first illumination light path (108), the first tube lens (222) of the optical system (104) is disposed between the at least one first optical element (105) and the second operating position (117) of the mirror assembly (114). The light sheet microscope (101) according to claim 8.

10. a ratio of a focal length (f) of the second optical element (113) to a focal length (f) of the second tube lens (223) is substantially equal to a ratio of a focal length (f) of the at least one first optical element (105) to a focal length (f) of the first tube lens (222); A light sheet microscope (101) according to any one of claims 7 to 9.

11. The mirror assembly (114) includes at least one adjustable mirror, the adjustable mirror comprising: the angle relative to the second illumination light path (108a), and - Position along the second illumination light path (108a) At least one of the following is configured to be adjusted: A light sheet microscope (101) according to any one of claims 1 to 10.

12. The optical system (104) includes a pair of adjusting mirrors, each of which has: the angle relative to the first illumination light path (108) and the second illumination light path (108a); and Positions along the first illumination light path (108) and the second illumination light path (108a) At least one of the following is configured to be adjusted: A light sheet microscope (101) according to any one of claims 1 to 11.

13. The first optical element (105) is a first objective lens (124), the second optical element (113) is a second objective lens (125), and the first objective lens (124) and the second objective lens (125) are Numerical aperture (NA), Working distance (wd), Pupil diameter (d), and ・Focal length (f) At least one of the following is different: A light sheet microscope (101) according to any one of claims 1 to 12.

14. The optical axis (126) of the first optical element (105) and the optical axis (127) of the second optical element (113) are - Substantially parallel to each other; - substantially antiparallel to each other, and - Substantially perpendicular to each other is oriented in one of A light sheet microscope (101) according to any one of claims 1 to 13.

15. The optical system (104) includes at least two scan mirrors configured to be adjustable in at least one of angle and position with respect to the first illumination optical path (108) and the second illumination optical path (108a); the scanning mirror is configured to scan the first sample volume (109) and the second sample volume (109a) with a light sheet (128) generated in the first sample volume (109) and the second sample volume (109a), A light sheet microscope (101) according to any one of claims 1 to 14.

16. The light sheet microscope (101) is an oblique microscope (102) or a SCAPE microscope (103). A light sheet microscope (101) according to any one of claims 1 to 15.

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