Microscope and microscopy method

Abstract

The invention concerns a microscope (1) comprising: - a light source (2) for illuminating a sample in an imaging position (3); - a detector (4) for capturing a signal from a sample in the imaging position (3); - a plurality of sample holders (5) for holding samples; - a translation device (7) for moving the plurality of sample holders (5) one at a time into the imaging position (3); and - a rotation device (6) for rotating a respective one of the plurality of sample holders (5) in the imaging position (3). The invention further concerns a corresponding microscopy method.

Description

[0001] Microscope and microscopy method

[0002] The present invention concerns a microscope and a microscopy method, in particular a fluorescence microscope and microscopy method, e . g . a confocal or light sheet microscope and microscopy method .

[0003] A microscope may use scattering, reflection, attenuation, absorption, or fluorescence to study the properties of organic or inorganic substances . In fluorescence microscopes , the specimen is illuminated with light of a speci fic wavelength ( or wavelengths ) , which is absorbed by f luorophores , causing them to emit light of di f ferent ( longer ) wavelengths . The illumination light may be separated from the emitted fluorescence , e . g . through the use of a spectral emission filter . One type of fluorescence microscope are epi fluorescence microscopes , where excitation of the fluorophore and detection of the fluorescence are done through the same light path ( i . e . through the ob j ective ) .

[0004] In confocal microscopy, the optical resolution and contrast is increased by means of using a spatial pinhole to block out-of- focus light in image formation . Capturing multiple two- dimensional images at dif ferent depths in a sample enables the reconstruction of three-dimensional structures within an obj ect ( optical sectioning) . As in traditional point-scanning confocal microscopy, only one point in the sample is illuminated at a time , 2D or 3D imaging requires scanning over a regular raster ( e . g . a rectangular pattern of parallel scanning lines ) in the specimen . One technique for achieving multipoint scanning confocal microscopy is the spinning disk technique . Spinningdisk confocal microscopes use a series of moving pinholes on a disc to scan spots of light . Since a series of pinholes scans an area in parallel , each pinhole is allowed to hover over a speci fic area for a longer amount of time , thereby reducing the excitation energy needed to illuminate a sample when compared to laser scanning microscopes . Decreased excitation energy reduces phototoxicity and photobleaching of a sample , often making it the preferred system for imaging live cells or organisms . To achieve a more complete or higher quality image , it may be advantageous to rotate the sample. Disadvantageously, present microscopes do not allow easy and swift imaging of multiple samples from multiple angles.

[0005] Another fluorescence microscopy technique particularly suitable for gentle long term live imaging of biological samples is light sheet fluorescence microscopy. Two types of light sheet systems are typically used in research. The first type allows live imaging of multiple samples but suffers from incomplete sample illumination, because a sample rotation mechanism is not implemented. Multiple samples are positioned in line inside a vessel-like chamber, which may only be translatable. In the second type, rotation is implemented but it allows imaging of only one sample, or a couple of samples stacked on top of each other in the same sample tube. In this system, the sample is loaded into a vertically positioned capillary.

[0006] A further option is a dual side illumination / dual side detection mechanism for multiple samples, however, this does not allow imaging from multiple angle (ranges) (see "Open top multi sample dual view light sheet microscope for live imaging of large multicellular systems", F. Moss et al., bioRxiv 2023.09.28.559925) . In another alternative light sheet microscopy configuration, multiple samples can be rotated (Bernardello, M., Gualda, E.J. & Loza-Alvarez, P. Modular multimodal platform for classical and high throughput light sheet microscopy. Sci Rep 12, 1969 (2022) ) . However, disadvantageously, in this set-up the sample loading is difficult, since all samples need to be positioned in line inside a single, horizontally oriented capillary.

[0007] It is an objective of the present invention to alleviate or overcome at least one of the problems of the prior art. In particular, it is an objective of the present invention to provide a microscope and microscopy method with a better performance, in particular that allow an easier, faster or better-quality imaging of multiple samples or a better scalability with regard to the number of samples.

[0008] This is achieved by a microscope comprising: - a light source for illuminating a sample in an imaging position;

[0009] - a detector for capturing a signal from a sample in the imaging position;

[0010] - a plurality of sample holders for holding samples ;

[0011] - a translation device for moving the plurality of sample holders one at a time ( i . e . sequentially) into the imaging position; characteri zed by

[0012] - a rotation device for rotating a respective one of the plurality of sample holders in the imaging position .

[0013] This is further achieved by a microscopy method, comprising the steps :

[0014] - providing a plurality of sample holders ;

[0015] - moving a first one of the plurality of sample holders into an imaging position;

[0016] - illuminating a sample in the first one of the plurality of sample holders in the imaging position and capturing a signal from the sample in the first one of the plurality of sample holders in the imaging position;

[0017] - rotating the first one of the plurality of sample holders in the imaging position;

[0018] - moving the first one of the plurality of sample holders out of the imaging position and moving a second one of the plurality of sample holders into the imaging position;

[0019] - illuminating a sample in the second one of the plurality of sample holders in the imaging position and capturing a signal from the sample in the second one of the plurality of sample holders in the imaging position;

[0020] - rotating the second one of the plurality of sample holders in the imaging position .

[0021] These steps are optionally conducted in the order as stated .

[0022] In this way, the microscope can hold multiple samples which can be studied individually . Each of the sample holders with a respective sample can be moved into the imaging position . (At least ) in the imaging position, the sample holder and with it the sample contained therein can be rotated . Thus , the sample in the imaging position can be examined from multiple directions . By moving the other ones of the plurality of sample holders into the imaging positions one at a time ( i . e . sequentially) , also the other samples can be examined from multiple directions . The microscope and method thus allow to examine a multitude of samples ef ficiently and with high image quality . These qualities can be maintained even if the number of samples and sample holders is scaled up, eliminating the need for multiple microscopy sessions to scale up the number of samples to be imaged with rotation . Also , the image quality does not need to be compromised versus the number of samples . Thanks to the possibility to image each sample from arbitrary angles , larger si zed samples can be examined . Further, in contrast to a system in which all samples need to be positioned in line inside a single , hori zontally oriented capillary, the partition of providing multiple sample holders each for holding one sample simpli fies sample retrieval and downstream analysis ( e . g . DNA or RNA extraction, proteomics , lysing and sorting) .

[0023] While the invention is applicable to microscopes and microscopy methods in general , it is particularly advantageous in the context of fluorescence microscopy, preferably light sheet fluorescence microscopy and confocal microscopy, particularly preferably confocal spinning disk microscopy .

[0024] The light source preferably provides a light beam directed at the sample or an area of a sample that is moved into the imaging position with the sample holder the sample is contained in . Capturing a signal from the sample refers to capturing a signal scattered by, reflected by, transmitted through and / or emitted ( e . g . by fluorescence ) from the sample . This may in particular be the ( attenuated, absorbed, scattered and / or reflected) light beam provided by the light source or light emitted from the sample , wherein the emittance is induced by the light beam provided by the light source .

[0025] There is provided for preferably at least 4 sample holders , particularly preferably at least 6 sample holders , even more preferably at least 8 sample holders . Each one of the sample holders in particular comprises a receptacle or container for a sample . The translation device allows to move sample holders and consequently the sample contained in the respective sample holders into and out of the imaging position, such that each sample holder can be individually positioned in the imaging position, for imaging of the sample contained therein . The respective sample holder in the imaging position is moved out of the imaging position as the next sample holder is moved into the imaging position . The translation device may in particular move the sample holders in at least one direction . This is not limited to mere translations , but may also comprise a rotational component , e . g . it can be achieved by swiveling in and out of the imaging position ( e . g . i f the sample holders are arranged in a circle ) . Optionally, the translation device is configured for adj usting a position of the sample holder within the imaging position in at least one direction, preferably in at least two non-parallel directions , even more preferably in three pairwise non-parallel directions . Optionally, the translation device is configured for moving the plurality of sample holders in at least one direction, preferably in at least two non-parallel directions , particularly preferably in three pairwise non- parallel directions ( e . g . in an x- , y- and z-direction, wherein each direction is non-parallel ( e . g . perpendicular ) with each other direction) . Thus , the sample cannot only be moved into and out of the imaging position, but its exact position / location within the imaging position may be adj ustable , such that di f ferent areas of the sample may be examined . Optionally, the actuator or motor is configured for driving the plurality of sample holders in at least one direction, preferably in at least two non-parallel directions , particularly preferably in three pairwise non-parallel directions . Optionally, the translation device simultaneously translated all of the plurality of sample holders . Optionally, the translation device comprises a motori zed stage , wherein the plurality of sample holders are mounted on the motori zed stage and / or the rotation device is mounted on the motori zed stage .

[0026] Optionally, the rotation device comprises an actuator or motor for ef fecting a rotation of the sample holder in the imaging position . The rotation device is configured at least for rotating the sample holder which is currently in the imaging position . I . e . , the rotation device is configured for rotating each sample holder at least when the respective sample holder is in the imaging position . The sample holder is in particular purely rotated and / or rotated around a rotation axis within the sample holder, in particular a central axis of the sample holder . The sample holder in the imaging position in particular remains in the imaging position as it is rotated by the rotation device . Optionally, the rotation device is configured for rotating the respective one of the plurality of sample holders in the imaging position by at least 180 ° , preferably by at least 270 ° , particularly preferably by at least 360 ° . The rotation device may rotate all sample holders at the same time . Optionally, the rotation device continuously (non-discreetly ) rotates the sample holder in the imaging position . Optionally, the rotation device is configured for rotating the sample holder in the imaging direction around a rotation axis and the translation device is configured for moving the plurality of sample holders one at a time into and out of the imaging position in a direction non-parallel to the rotation axis , in particular perpendicular to the rotation axis .

[0027] The method preferably comprises the step : providing a sample in more than one of the plurality of sample holders , i . e . at least in the first one of the plurality of sample holders and in the second one of the plurality of sample holders . For the sake of simplicity, the first one of the plurality of sample holders is referred to as the first sample holder and the second one of the plurality of sample holders is referred to as the second sample holder in the following . The method may comprise for one or more further ones of the plurality of sample holders the steps , one at a time :

[0028] - moving a respective one of the plurality of sample holders currently in the imaging position out of the imaging position and moving the respective further one of the plurality of sample holders into the imaging position;

[0029] - illuminating a sample of the respective further one of the plurality of sample holders in the imaging position and capturing a signal from the sample in the respective further one of the plurality of sample holders in the imaging position;

[0030] - rotating the respective further one of the plurality of sample holders in the imaging position . Optionally, the method comprises multiple instances of the rotation and the illumination and capturing steps of the respective sample holders in the imaging position . Optionally, the respective sample in the first sample holder and / or the second sample holder and / or one or more further ones of the plurality of sample holders may also be illuminated and the signal captured during and / or after rotating the respective sample holder in the imaging position . The first sample holder and / or the second sample holder and / or one or more further ones of the plurality of sample holders may be rotated multiple times in the imaging position, wherein the respective sample may be illuminated and the signal captured during and / or after some or after every instance of rotation .

[0031] Optionally, the rotation device is configured for simultaneously rotating all ( each one ) of the plurality of sample holders . This allows easier determination of the rotational status of each one of the plurality of sample holders and allows a less complex setup . Optionally, all of the plurality of sample holders are simultaneously rotated by the same absolute value of the angle of rotation . E . g . every other sample holder may be rotated in the opposite direction but by the same absolute value of the angle . Optionally, all of the plurality of sample holders are simultaneously rotated by the same angle .

[0032] Optionally, each one of the plurality of sample holders comprises a coupling element , wherein the rotation device is coupled ( directly or indirectly) with the coupling element of each of the plurality of sample holders for ef fecting a rotation of the plurality of sample holders . I . e . rotation of the sample holders can be ef fected via the respective coupling element . The receptacle of each one of the sample holders is in particular connected to the respective coupling element in a torque-proof way .

[0033] Optionally, the coupling element of each of the plurality of sample holders is a gear ( circular gear, including cogwheels ) and the coupling elements of adj acent ones of the plurality of sample holders ( i . e . pairwise ) engage with each other, such that rotation of the coupling element of one of the plurality of sample holders ef fects rotation of the coupling elements of the other ones of the plurality of sample holders , wherein the rotation device engages with the coupling element of the one of the plurality of sample holders . In this way, rotation of all sample holders is achieved in a structurally simple way . In particular, rotation of each individual one of the plurality of sample holders ef fects rotation of all of the plurality of sample holders .

[0034] Optionally, the coupling element of each of the plurality of sample holders is a gear ( circular gear, including cogwheels ) and the rotation device comprises a gear rack ( linear gear ) , which engages with the respective gear of each of the plurality of sample holders . I . e . , rotation of the sample holders may be ef fected by a rack and pinion system . Thus , movement of the gear rack ef fects rotation of all of the plurality of sample holders ( in particular in the same direction) . The rotation device may be configured for moving the gear rack linearly .

[0035] Optionally, the rotation device comprises a belt ( e . g . a rubber band) in contact with the coupling element of each one of the plurality of sample holders ( such that driving of the belt rotates the coupling elements and consequently sample holders ) . In particular, the belt is formed as a loop . In particular, the rotation device is configured for driving the belt . In particular, the rotation device comprises a belt drive .

[0036] Optionally, the translation device is configured for translating the rotation device simultaneously with the plurality of sample holders . In particular, the rotation device is mounted on the translation device .

[0037] Optionally, the translation device is configured for adj usting a position of the respective one of the plurality of sample holders in the imaging position in at least two non-parallel directions , preferably in three pairwise non-parallel directions . Thus , di f ferent areas / depths of the sample can be examined .

[0038] Optionally, each of the plurality of sample holders comprises a tubular cavity for receiving a sample . Optionally, the sample holder comprises an Eppendorf-like tube or a capillary or a cuvette . Alternatively, sample holders could also hold the sample in a hanging configuration .

[0039] Optionally, in an operating position of the microscope an axial direction of the tubular cavity of each of the plurality of sample holders is non-hori zontal , in particular substantially vertical . This makes sample loading easier .

[0040] Optionally, the rotation device is configured for rotating the respective one of the plurality of sample holders in the imaging position about a rotation axis parallel to an axial direction of the tubular cavity of the respective one of the plurality of s amp le holders .

[0041] Optionally, the plurality of sample holders are arranged along a line , optionally a straight line . The plurality of sample holders may be arranged along a closed curve , such as a circle .

[0042] Optionally, the microscope is a light sheet fluorescence microscope , wherein the light source is for illuminating the sample in the imaging position with a light sheet and the detector is for capturing a fluorescence signal emitted from the sample in the imaging position .

[0043] Optionally, the microscope is configured for conducting the microscopy method according to any of the embodiments described in this disclosure . Optionally, the microscope comprises a control unit for controlling the microscope for conducting the method according to any of the embodiment mentioned in this disclosure .

[0044] Regarding the microscopy method, optionally, each sample holder holds at maximum one sample . This simpli fies sample loading and examination . Optionally, each sample holder holds exactly one sample . Optionally, each sample is a biological sample .

[0045] Optionally, upon ( i . e . while ) rotating the first one of the plurality of sample holders in the imaging position and upon rotating the second one of the plurality of sample holders in the imaging position, respectively, all of the plurality of sample holders are rotated ( i . e . simultaneously) . Optionally, the method is a light sheet fluorescence microscopy method, wherein the samples are respectively illuminated with a light sheet and fluorescent signals emitted from the samples are respectively captured .

[0046] Optionally, the method is a confocal microscopy method, in particular a confocal spinning disk microscopy method .

[0047] Preferably each of the plurality of sample holders is purely rotated and / or each of the plurality of sample holders is rotated around a rotation axis within the respective sample holder ( in particular simultaneously with the rotation of the sample holder in the imaging position) , in particular around a central axis of the respective sample holder . Preferably the sample holders are arranged along a line , preferably a line that at least in part is non-parallel to the rotation axis of any of the sample holders . Preferably the sample holders are arranged along a hori zontal line in the operating position of the microscope . Preferably in the operating position of the microscope the axis of illumination and the axis of detection intersect only the sample of the respective sample holder of the plurality of sample holders in the imaging position . Preferably at least the rotation axis of the respective sample holder of the plurality of sample holders in imaging position is such that the orientation of an axis of rotational symmetry of the sample holder relative to a static detection axis is irrespective of the rotation of the sample holder . Preferably, an axial direction of the tubular cavity of each of the plurality of sample holders is parallel to the rotation axis . Preferably in the operating position of the microscope at least the respective sample holder of the plurality of sample holders in imaging position is accessible and / or can be displaced and / or replaced . Preferably in the operating position of the microscope at least the tubular cavity of the respective sample holder of the plurality of sample holders in imaging position is accessible . Preferably in the operating position of the microscope at least the respective sample holder of the plurality of sample holders in imaging position is accessible from a direction parallel to the rotation axis of said sample holder . Preferably each of the plurality of sample holders comprises a coupling element . Preferably the rotation device is coupled with the coupling element of a number of sample holders of the plurality of sample holders , wherein the number is lower than the total number of sample holders ; even more preferably the number is one . Preferably the coupling of the rotation device of a number of the plurality of sample holders ef fects a rotation of the plurality of sample holders . Preferably in the operating position of the microscope the distance between each of the plurality of sample holders and its neighboring sample holder ( s ) along a plane normal to the rotation axis is equidistant . Preferably, the plurality of sample holders are arranged spaced apart from each other in a direction which is non-parallel , in particular perpendicular, to a rotation axis around which the respective one of the plurality of sample holders is rotated in the imaging position ( in particular non-parallel to the rotation axes around which each of the plurality of sample holders are rotated simultaneously) .

[0048] Optionally, the rotation device comprises a ( rotating) actuator or a ( rotating) motor for each sample holder, wherein each actu- ator / motor is for ef fecting a rotation of the respective sample holder . I . e . , there can be an individual actuator / motor provided for ( and coupled with) each individual sample holder . In this way, only the sample holder in the imaging position can be rotated or all sample holders can be rotated simultaneously ( depending on the driving of the motors / actuators ) .

[0049] By way of example , the disclosure is further explained with respect to some selected embodiments shown in the drawings . However, these embodiments shall not be considered limiting for the disclosure . In the drawings :

[0050] Fig . 1 schematically illustrates a first embodiment of a microscope according to the present disclosure in a top view;

[0051] Fig . 2 schematically illustrates a second embodiment of the microscope according to the present disclosure in a top view;

[0052] Fig . 3 schematically illustrates the second embodiment of the microscope in an axonometric proj ection;

[0053] Fig . 4 schematically illustrates a third embodiment of the microscope according to the present disclosure in a top view; Fig . 5A schematically illustrates a first mechanism for rotating multiple sample holders at the same time , in an exploded view;

[0054] Fig . 5B schematically illustrates a second mechanism for rotating multiple sample holders at the same time , in an exploded view;

[0055] Fig . 5C schematically illustrates a third mechanism for rotating multiple sample holders at the same time .

[0056] Fig . 1 schematically illustrates a first embodiment of a microscope 1 according to the present disclosure in a top view . In this embodiment , the microscope 1 is a confocal microscope . The microscope 1 comprises a light source 2 for illuminating a sample in an imaging position 3 and a detector 4 for capturing a signal from the sample in the imaging position 3 . The detector 4 may e . g . comprise a photomultiplier tube , an avalanche photodiode , a photodetector and / or a camera . A light beam emitted by the light source 2 is directed to a beam splitter 16 and focused onto a sample in the imaging position 3 by an obj ective 17 . Light , in particular fluorescence light , emitted by the sample in the imaging position 3 is collected by the same obj ective 17 and via the beam splitter 6 directed to the detector 4 .

[0057] The microscope 1 comprises a plurality of sample holders 5 , each for holding a sample . The plurality of sample holders 5 are arranged along a straight line . The microscope 1 further comprises a translation device 7 for moving the plurality of sample holders 5 one at a time into the imaging position 3 . In particular, the translation device 7 is for moving the plurality of sample holders simultaneously at least in one direction 12 ( the up-down direction in the drawing plane of Fig . 1 ) . In this way, each of the sample holders 5 can be moved into and out of the imaging position 3 , such that each sample can be individually examined . The translation device 7 in particular moves the sample holders 5 at least in the direction 12 perpendicular to the direction of light illumination and sampling .

[0058] The microscope 1 further comprises a rotation device 6 for rotating a respective one of the plurality of sample holders 5 in the imaging position 3 . In this embodiment , the rotation device 6 is mounted on the translation device 7 ( and therefore translated together with the sample holders 5 ) and is configured for rotating all of the sample holders 5 at the same time . In summary, the microscope 1 allows to examine the respective sample in each of the sample holders 5 from multiple angles , in particular from continuous ranges of angles .

[0059] The rotation device 6 is configured for rotating the sample holders 5 about a rotation axis 15 (not shown in Fig . 1 ) , which is perpendicular to the drawing plane in Fig . 1 . The translation device 7 translates the sample holders at least in the direction 12 perpendicular to the rotation axis 15 .

[0060] Each of the plurality of sample holders 5 comprises a tubular cavity 13 (not shown in Fig . 1 ) for receiving the respective sample . In an operating position of the microscope 1 an axial direction 14 of the tubular cavity 13 of each of the plurality of sample holders 5 is substantially vertical ( and parallel to the rotation axis 15 ) and the translation device 7 translates the sample holders at least in the direction 12 perpendicular to the axial direction 14 (not shown in Fig . 1 ) .

[0061] Figs . 2 and 3 schematically illustrate a second embodiment of the microscope 1 , which is also a confocal microscope . Fig . 2 schematically shows a top view and Fig . 3 an axonometric proj ection . The second embodiment di f fers from the first embodiment in that the translation device is configured for translating the plurality of sample holders 5 in three pairwise perpendicular directions 12 . In this way, the sample holders 5 can be moved into and out of the imaging position 3 ; at the same time , the exact position of the sample to be examined within the imaging position 3 can be adj usted . Thus , di f ferent areas / depths of the sample in the respective sample holder 5 in the imaging position 3 can be examined .

[0062] Fig . 3 also shows the sample holders 7 each comprising a tubular cavity 13 , in particular an Eppendorf-like tube , and illustrates their axial direction 14 as well as the rotation axis 15 (which are parallel and identical ) . Fig . 4 schematically illustrated a third embodiment of the microscope 1 in a top view . This embodiment di f fers from the first embodiment in that the microscope 1 is a light sheet fluorescence microscope . Therefore , in this embodiment , the light source 2 provides a light sheet , which is focused onto the sample in the imaging position 3 by a first obj ective 17a .

[0063] A second obj ective 17b collects the beam emitted from the sample by fluorescence (which is substantially perpendicular to the illumination beam) and directs it to the detector 4 . The translation device 7 moves the plurality of sample holders 5 in a direction 12 , which is at an angle of e . g . 45 ° to the illumination beam and the emission beam .

[0064] Figs . 5A to 5C schematically illustrate three embodiments of the rotation mechanism, which could e . g . be used in the first , second and third embodiment of the microscope 1 .

[0065] In the first embodiment of the rotation mechanism shown in Fig .

[0066] 1 , each of the sample holders 5 comprises a coupling element 8 , wherein the rotation device 6 is coupled with the coupling element 8 of each of the sample holders 5 for ef fecting a rotation of the sample holders 5 . In particular, the rotation device 6 directly engages with one of the coupling elements 8 , and the coupling elements 8 of adj acent sample holders 5 engage with each other, such that the rotation device 6 is directly or indirectly coupled to the coupling element 8 of each sample holder 5 . In this embodiment , the rotation mechanism is in particular implemented as a spur gear . The coupling element 8 of each sample holder 5 is a gear 9, and the coupling elements 8 of adj acent ones of the plurality of sample holders 5 engage with each other, such that rotation of the coupling element 8 of one of the plurality of sample holders 5 ef fects rotation of the coupling elements 8 of the other ones of the plurality of sample holders 5 , wherein the rotation device 6 engages with the coupling element 8 of the one of the plurality of sample holders 5 .

[0067] In the second embodiment of the rotation mechanism shown in Fig .

[0068] 2 , the rotation mechanism is a rack and pinion mechanism . The coupling element 8 of each of the plurality of sample holders 5 is a gear 9 and the rotation device 6 comprises a gear rack 10 , which engages with the gear 9 of each of the plurality of sample holders 5 . I . e . in this embodiment , the rotation device 6 directly engages with the coupling element 8 of each sample holder 5 .

[0069] In the third embodiment of the rotation mechanism shown in Fig . 3 , the rotation device 6 comprises a belt 11 in contact with the coupling element 8 of each of the sample holders 5 . Thus , driving the belt 11 rotates each sample holder 5 . The belt 11 is in the form of a loop .

[0070] While in the first embodiment of the rotation mechanism, each other sample holder 5 rotates in the opposite direction (but by substantially the same absolute value of the angle ) , in the second and third embodiment of the rotation mechanism, each sample holder 5 rotates in the same direction ( and by substantially the same angle ) .

Claims

Claims :

1. Microscope (1) comprising:- a light source (2) for illuminating a sample in an imaging position ( 3 ) ;- a detector (4) for capturing a signal from a sample in the imaging position (3) ;- a plurality of sample holders (5) for holding samples;- a translation device (7) for moving the plurality of sample holders (5) one at a time into the imaging position (3) ; characterized by- a rotation device (6) for rotating a respective one of the plurality of sample holders (5) in the imaging position (3) .

2. Microscope (1) according to claim 1, wherein the rotation device (6) is configured for simultaneously rotating all of the plurality of sample holders (5) .

3. Microscope (1) according to claim 2, wherein each of the plurality of sample holders (5) comprises a coupling element (8) , wherein the rotation device (6) is coupled with the coupling element (8) of each of the plurality of sample holders (5) for effecting a rotation of the plurality of sample holders (5) .

4. Microscope (1) according to claim 3, wherein the coupling element (8) of each of the plurality of sample holders (5) is a gear (9) and the coupling elements (8) of adjacent ones of the plurality of sample holders (5) engage with each other, such that rotation of the coupling element (8) of one of the plurality of sample holders (5) effects rotation of the coupling elements (8) of the other ones of the plurality of sample holders(5) , wherein the rotation device (6) engages with the coupling element (8) of the one of the plurality of sample holders (5) .

5. Microscope (1) according to claim 3, wherein the coupling element (8) of each of the plurality of sample holders (5) is a gear (9) and the rotation device (6) comprises a gear rack (10) , which engages with the gear (9) of each of the plurality of sample holders (5) .

6. Microscope (1) according to claim 3, wherein the rotation device (6) comprises a belt (11) in contact with the coupling element (8) of each of the plurality of sample holders (5) .

7. Microscope (1) according to any one of the previous claims, wherein the translation device (7) is configured for translating the rotation device (6) simultaneously with the plurality of sample holders (5) .

8. Microscope (1) according to any one of the previous claims, wherein the translation device (7) is configured for adjusting a position of the respective one of the plurality of sample holders (5) in the imaging position (3) in at least two non-parallel directions (12) , preferably in three pairwise non-parallel directions ( 12 ) .

9. Microscope (1) according to any one of the previous claims, wherein each of the plurality of sample holders (5) comprises a tubular cavity (13) for receiving a sample.

10. Microscope (1) according to claim 9, wherein in an operating position of the microscope (1) an axial direction (14) of the tubular cavity (13) of each of the plurality of sample holders(5) is substantially vertical.

11. Microscope (1) according to any one of claims 9 and 10, wherein the rotation device (6) is configured for rotating the respective one of the plurality of sample holders (5) in the imaging position (3) about a rotation axis (15) parallel to an axial direction (14) of the tubular cavity (13) of the respective one of the plurality of sample holders (5) .

12. Microscope (1) according to any one of the previous claims, wherein the plurality of sample holders (5) are arranged along a line, optionally a straight line.

13. Microscope (1) according to any one of the previous claims, wherein the microscope (1) is a light sheet fluorescence microscope (1) , wherein the Microscope (1) is for illuminating the sample in the imaging position (3) with a light sheet and thedetector (4) is for capturing a fluorescence signal emitted from the sample in the imaging position (3) .

14. Microscopy method, comprising the steps:- providing a plurality of sample holders (5) ;- moving a first one of the plurality of sample holders (5) into an imaging position (3) ;- illuminating a sample in the first one of the plurality of sample holders (5) in the imaging position and capturing a signal from the sample in the first one of the plurality of sample holders (5) in the imaging position (3) ;- rotating the first one of the plurality of sample holders (5) in the imaging position (3) ;- moving the first one of the plurality of sample holders (5) out of the imaging position (3) and moving a second one of the plurality of sample holders (5) into the imaging position (3) ;- illuminating a sample in the second one of the plurality of sample holders (5) in the imaging position (3) and capturing a signal from the sample in the second one of the plurality of sample holders (5) in the imaging position (3) ;- rotating the second one of the plurality of sample holders (5) in the imaging position (3) .

15. The method according to claim 14, wherein each sample holder holds at maximum one sample.

16. The method according to any one of claims 14 and 15, wherein upon rotating the first one of the plurality of sample holders(5) in the imaging position (3) and upon rotating the second one of the plurality of sample holders (5) in the imaging position (3) , all of the plurality of sample holders (5) are rotated.

17. The method according to any one of claims 14 to 16, wherein the method is a light sheet fluorescence microscopy method, wherein the samples are respectively illuminated with a light sheet and fluorescent signals emitted from the samples are respectively captured.18 . The method according to any one of claims 14 to 16 , wherein the method is a confocal microscopy method, in particular a confocal spinning disk microscopy method .

Images