Microscopy arrangement with microscope and method of operation thereof

Abstract

The invention relates to a method of operating a microscopy arrangement (M) having a first microscope (1) and at least one other microscope (3), wherein the microscopes (1, 3) each have an optical axis (2.1, 4.1) and the optical axes (2.1, 4.1) do not coincide. The method according to the invention comprises the steps A to F, wherein a three-dimensional reference coordinate system is set; a carrier device (6) embodied to receive and hold a sample carrier (7) is introduced into a sample plane (9) of the first microscope (1) intersected by the optical axis (2.1) and onto the optical axis (2.1) of the first microscope (1); a reference point (11) is set on the optical axis (2.1); the carrier device (6) is delivered to the other microscope (3), wherein the current coordinates of the reference point (11) are continuously captured and compared with the coordinates of the optical axis (4.1) of the other microscope (3); the reference point (11) is brought onto the second optical axis (4.1). Furthermore, the invention relates to a microscopy arrangement (M).

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201810932399.5, with the filing date of August 15, 2018 and the title of “Microscopy arrangement with microscope and method of operation thereof”. TECHNICAL FIELD

[0002] The present invention relates to a method of operating a microscopy arrangement with a first microscope and at least one other microscope, and to a microscopy arrangement. BACKGROUND

[0003] If a sample to be examined is examined by different microscopes and / or different microscopy methods, it is problematic to find (retrace) the region of interest (ROI) of the sample to be examined when the sample is transferred from one microscope to the other microscope.

[0004] In the workflow of a microscopy examination of a sample denoted as “conventional”, the sample is placed on a suitable object carrier and set up with respect to the optical axis of the microscope. Subsequently, an overview image of the sample can be recorded first by an ocular, at low magnification and large field of view (FoV), if required, in order to identify the region of interest (ROI) of the sample. Subsequently, the sample can be examined using different examination methods, such as (fluorescent) contrast methods, DIC (differential interference contrast), polarization contrast, phase contrast, Hoffmann modulation contrast, dark field illumination, etc.

[0005] If different examination methods should be applied using only one microscope system, this one microscope system must have an ocular beam path and at least one camera beam path. In addition, there is a need for, for example, reflected and transmitted light illumination, objective exchange means, filter exchange means, pupil engagement(s), components for, for example, DIC (differential interference contrast), etc.

[0006] Arms for microscopes can facilitate an inverted or upright arrangement of the microscope. In order to allow the application of different examination methods, the arm needs further interfaces, e.g. for laser scanning microscopy (LSM), total internal reflection fluorescence microscopy (TIRF microscopy), fluorescence methods (e.g. fluorescence recovery after photobleaching; FRAP), Vivatome, spinning disk, units for generating structured illumination (e.g. Apotome etc.), or combinations thereof. In these systems, the sample remains substantially at the same position and so the same sample region can be examined using different objectives and / or methods. The disadvantage here is that many interfaces or many optical paths and switching devices have to be kept available both on the illumination side and on the detection side in order to be able to operate all selection items (methods, modes). Furthermore, the conventional arm always has to have an objective exchange device, e.g. an objective revolver, in order to be able to operate different modes (e.g. change between providing overview images and images with high resolution). Naturally, such an objective exchange device impairs the stability of the optical system (so-called drift) with respect to an independent, fixedly assembled objective.

[0007] In contrast to this are dedicated microscopy systems with a specified objective arrangement, which are very specifically designed for one examination method, such as SPIM. These do not allow objective exchange, for example. Even alternative contrast methods such as DIC, polarization contrast etc. are only possible with these objective arrangements in a very limited range.

[0008] If the same sample is examined using both a dedicated examination method and, for example, a conventional microscopy, it is necessary to adopt the associated method (as is known from the combination of light and electron microscopy).

[0009] Here, the problem arises of finding the ROI in the sample if it is transported from the dedicated system to the conventional microscope and vice versa. This process is also called "shuttle and find". This can be carried out either using a very precisely manufactured sample frame, which forms a reference coordinate system, or by means of a marked sample container, which has, for example, a listed grid, on the basis of which the desired position can be found in the respective other microscope system. However, this process is time-consuming and requires intensive work and does not ensure a quick and repeatable exchange between the modes in a satisfactory manner.

[0010] The prior art has disclosed microscopy arrangements which are implemented to examine samples with different examination methods.

[0011] DE 10 2012 014 768 B4 has already described a microscope with a first and a second imaging optical unit which are arranged spaced apart from one another. They serve for imaging a first and a second object field into a first and a second image field using a first illumination radiation. In addition, a third imaging optical unit is present which images a third object field into a third image field using a second illumination radiation. Here, the first, second and third object fields are arranged in an object plane, and the first and second image fields are arranged to overlap in an image plane. The wavelength of the first illumination radiation is longer than the wavelength of the second illumination radiation. The centers of the object fields lie on a straight line, and the first and second object fields surround the third object field.

[0012] WO 2014 / 173547 relates to a microscopy method and apparatus for multi-dimensional positioning of an object of interest. Here, at least two reference markers of a reference coordinate system are used in order to establish a position of the object in the reference coordinate system. The thus established position can be used by a second microscope in order to reposition the object.

[0013] A press release of the Fraunhofer Institute for Photonic, Microsystems and Optics (issued on January 24, 2014) has already disclosed a microscopy arrangement of different microscopes operated by a robot. The robot has a central control unit and transfers the respective sample to the different microscopes with positioning accuracy. SUMMARY

[0014] The present invention is based on the object of proposing an alternative for checking a sample with different checking methods, wherein the region of interest of the sample is prompted to be precisely repositioned in case of a change between the checking methods. At the same time, a device type option should be proposed in order to ensure a high repositioning accuracy.

[0015] The object is achieved by the method with the features described below with respect to the precise repositioning option, and according to the description below with respect to the microscopy arrangement. Advantageous developments are described below.

[0016] The method is designed to operate a microscopy arrangement with a first microscope and at least one other microscope, wherein each of the microscopes has an optical axis and the optical axes do not coincide.

[0017] The method according to the invention comprises the steps A to F. Other intermediate steps can be carried out in other configurations of the method.

[0018] In step A, a three-dimensional reference coordinate system is set, in which the coordinates of the optical axis range of the first microscope and the coordinates of the optical axis range of the other microscopes are known or established. The known or established coordinates are stored and used as reference axes. As an example, the reference coordinate system is set by means of the optical axis of the first microscope, which coincides with one of the axes of the reference coordinate system, and a point at which the optical axis intersects with a front lens of the objective of the first microscope, for example, which point is set as a defined point of the reference coordinate system, for example, as zero point or origin.

[0019] If the microscopy arrangement has exactly two microscopes, the other microscope is the second present microscope. In contrast, if there are more than two microscopes, the other microscope is the microscope which is to be used next. This does not necessarily have to be the microscope which is spatially closest to the first microscope.

[0020] The coordinates of the range of one of the optical axes can be captured and stored in a manner known to the person skilled in the art. As an example, the coordinates of the range are represented as a vector (point and direction).

[0021] When the first objective is in the working position in the first microscope, the first optical axis is given by the first objective, and so the image data of the sample are captured or can be captured by the first objective. The second optical axis is thus given by the other objective of the other microscope which is in the working position.

[0022] In step B, a carrier device which is implemented to receive and hold the sample carrier is introduced into the sample plane of the first microscope which is intersected by the first optical axis and onto the optical axis of the first microscope.

[0023] For the purpose of carrying out the method, the object to be imaged, for example the sample, is preferably arranged in or on the sample carrier.

[0024] The sample plane is an essential feature of the respective microscope or of the objective currently used and is preferably given by the respective focal plane of the microscope.

[0025] Step C comprises setting a reference point which coincides with the optical axis and establishing the coordinates of the set reference point.

[0026] In step D, the carrier device is delivered to the other microscope, in which the current coordinates of the reference point are continuously captured and compared with the coordinates of the optical axis of the other microscope.

[0027] In step E, a control command is generated depending on the difference established by the comparison between the current coordinates of the reference point and the coordinates of the optical axis of the other microscope, with which control command the further delivery movement of the carrier device is controlled in step F in such a way that the carrier device is positioned in such a way that the reference point coincides with the other optical axis.

[0028] The core idea of the invention consists of the continuous capturing of the coordinates of the carrier device and, optionally, of the coordinates of the sample. This ensures that the sample never leaves the reference coordinate system and that every region of the sample can be retrieved and targeted without the methods within the meaning of special marking and / or "shuttling and finding".

[0029] The continuous capturing of the coordinates is provided if the coordinates are captured at very short intervals, in particular several times per second. As an example, the capturing is carried out at a frequency of at least five, ten, twenty, fifty, one hundred or one thousand captures per second.

[0030] As an example, the frequency of the continuous capturing can be selected and adjusted depending on the displacement speed of the carrier device. Thus, at low displacement speeds, a low capturing frequency can also be selected, and vice versa.

[0031] The continuous capturing can also be achieved by means of a counting movement unit of the carrier device, for example a rotation-dependent rotation of an actuator and / or a direction-dependent step of a stepper motor, which is captured, evaluated and, if necessary, stored.

[0032] This idea is particularly advantageous for combining microscopy methods which require very specific optical or mechanical arrangements with conventional methods. Thus, for example, a fixed and thus very stable optical unit has a high flexibility in the selection of the examination method and at the same time the advantage of a high accuracy of the repositioning is achieved.

[0033] Furthermore, the time required for changing the sample between various examination methods is limited only to the distance to be travelled and to the displacement speed of the mobile carrier device. The time required for the change is thus known in advance and is standardizable, which in turn serves to improve the reproducibility of the experiment.

[0034] In an advantageous further configuration of the method, the coordinates of the reference point are compared in step D with setpoint coordinates of a target point of the object on the optical axis of the further microscope. In step E, a control command is generated depending on the difference established by comparing the current coordinates of the reference point and the coordinates of the target point, and in step F the further delivery movement of the carrier device is controlled by means of this control command, so that the carrier device is positioned in such a way that the reference point coincides with the target point. Thus, the carrier device and possibly the sample located thereon are also in a uniquely determined position relative to each other in the direction of the optical axis of the further microscope, which is usually referred to as the z direction.

[0035] In a further possible configuration of the method, image data of the object region are captured in step C and a reference point is set within the captured object region. The coordinates of the object region are known or established and stored. Image data of a further object region are captured by means of the further microscope after step F, wherein the reference point is located in the further object region.

[0036] Optionally, before or during the capturing of the image data, the microscope used for capturing the image is focused along the optical axis.

[0037] The above configurations of the method can be combined with each other.

[0038] In the context of a device-based design, the object is achieved by a microscopy arrangement comprising a first microscope and at least one other microscope, wherein each of the microscopes has an optical axis and the optical axes do not coincide. A carrier device is present which receives and holds a sample carrier. Furthermore, a computer unit is present which is implemented to set up a three-dimensional reference coordinate system, wherein the coordinates of the extent of the optical axis of the first microscope and the coordinates of the extent of the other optical axis of the other microscope are known and stored, or wherein the coordinates of the extent of the optical axis of the first microscope and the coordinates of the extent of the other optical axis of the other microscope, as established by a present measuring member, are stored and treated as reference axes. A transport device is present to deliver the carrier device into one of the optical axes. A coordinate measuring device is present by which the current coordinates of the carrier device are continuously captured or can be captured. Furthermore, the computer unit is also implemented to compare the current coordinates of the carrier device captured by the coordinate measuring device with the coordinates of the optical axes of the other microscopes, wherein a comparison between the coordinates of the reference point set on the optical axis of the first microscope and the coordinates of the optical axes of the other microscopes is achieved. In order to generate control commands, a control unit is present which is implemented to generate control commands depending on the difference established by the comparison between the current coordinates of the reference point and the coordinates of the optical axes. Furthermore, the control unit is implemented to control the delivery movement of the carrier device depending on the control commands, so that the carrier device is positioned in such a way that the reference point coincides with the other optical axis.

[0039] As an example, the reference coordinate system can be configured substantially by the following: a spatial point actually present is defined as a certain point of the reference coordinate system and serves as a reference for the optical axes and all affected movements (change of coordinates) and position coordinates of the technical elements. It can also be that the current relative positions of at least two technical elements, at least three technical elements, which define the spatial relationship (better) in the microscopy arrangement, serve as reference coordinates of the reference coordinate system. After setting up the reference coordinate system, the optical axes serving as reference axes are determined and then serve as the necessary reference variables, but not necessarily only as reference variables.

[0040] Continuous capturing of the coordinates can be achieved, for example, by capturing and evaluating the respective realized delivery path of the employed drive, i.e. using a path measuring system in the actuation system, and / or continuous capturing of reference markers (traces) of the carrier device.

[0041] As an example, the sample carrier is a cover glass, a plate or a petri dish. Advantageously, it consists of a material such as glass or transparent plastic, which is transparent to the illumination radiation employed and / or to the detection radiation to be captured. The sample carrier is provided as a sample plane, in which the sample to be examined is arranged or can be arranged. Preferably, the sample plane extends horizontally, but it can also be provided in other spatial planes in other embodiments of the microscopy arrangement.

[0042] At least one microscope of the microscopy arrangement can have a plurality of objectives, which can be pivoted or inserted into the respective optical axis by means of an objective exchange device, for example by means of an objective rotator or a linear slide. The reduced stability is offset by an increased flexibility in terms of possible magnifications and / or examination methods.

[0043] It can also be that at least one of the microscopes of the microscopy arrangement has a plurality of objectives which are simultaneously directed together into the sample plane.

[0044] In possible embodiments, the first microscope and / or the further microscopes each have an illumination objective and a detection objective, the optical axes of which are perpendicular to one another and are directed into the sample plane. The illumination objective can be embodied to produce a light sheet which intersects the sample plane.

[0045] In other embodiment options, the microscopy arrangement according to the application can have an embodiment of the inverted type. Here, the optical axes are directed onto or into the sample through the sample carrier employed and / or through the carrier device.

[0046] In other embodiments of the microscopy arrangement, the optical axes each comprise an angle which is different from zero with the sample plane and the normal to the sample plane.

[0047] The above options can be combined with one another. As an example, a microscope embodied as a light sheet microscope can be present. The microscope can be embodied as a right-side-up alignment or as an inverted alignment.

[0048] In other embodiments of the microscopy arrangement, there can be present, next to at least two microscopes, a work space having a work position. A work axis can extend through the work position. The coordinates of the work position and, optionally, the coordinates of the work axis are known within a reference coordinate system. According to the method according to the application, the carrier device can be delivered to the work position or to the work axis.

[0049] As an example, the work space can additionally or exclusively be used for the processing or non-optical examination of the sample, and can be embodied and set, for example, as a filling station which changes the buffer or fills sample containers by means of a pipetting machine. BRIEF DESCRIPTION OF DRAWINGS

[0050] The application is explained in more detail below on the basis of exemplary embodiments and the drawings. In the drawings:

[0051] Figure 1a a schematic diagram of a first exemplary embodiment of a microscopy arrangement according to the present application is shown in a side view;

[0052] Figure 1b a schematic diagram of a first exemplary embodiment of a microscopy arrangement according to the present application is shown in a top view;

[0053] Figure 2 a schematic diagram of a second exemplary embodiment of a microscopy arrangement according to the present application is shown;

[0054] Figure 3 a schematic diagram of a third exemplary embodiment of a microscopy arrangement according to the present application is shown; and

[0055] Figure 4 a schematic diagram of one exemplary embodiment of a microscope for light sheet microscopy is shown. DETAILED DESCRIPTION

[0056] An exemplary embodiment of a microscopy arrangement M is shown in the following in a schematic and exemplary manner in an inverted type embodiment. As basic elements of the microscopy arrangement M there are a first microscope 1 with a first objective 2 and a first optical axis 2.1 and a further microscope 3 with a further objective 4 and a second optical axis 4.1, wherein the optical axes 2.1 and 4.1 extend parallel to each other and do not coincide Figure 1a ).

[0057] Figure 1a There is a carrier device 6 on which a sample carrier 7 is placed. A sample 8 to be examined is located on the sample carrier 7. A sample plane 9 is provided by the upper side of the sample carrier 7, which extends horizontally in the exemplary embodiment and also serves as a reference surface below.

[0058] In other embodiments of the present application, the sample plane 9 can be defined in the plane of the carrier device 6, so that possible inhomogeneities of the sample carrier 7 do not have to be taken into account.

[0059] The carrier device 6 is located in a first observation, examination or measurement position PI (marked with an arrow; in the following referred to as measurement position), at which the sample 8 is located on the first optical axis 2.1 and at which at least one image of the sample 8 or a region of the sample 8 is captured or can be captured by the first objective 2 when the examination method is carried out.

[0060] Furthermore, there is a computing unit 13 which is implemented to set a three- dimensional reference coordinate system with the axes x, y and z. The coordinates of the range of the first optical axis 2.1 of the first microscope 1 and the coordinates of the range of the further optical axis 4.1 of the further microscope 3 are known and stored in the computing unit 13. In other embodiment options, the ranges of the optical axes 2.1 and 4.1 are established by means of the measuring means present, in particular by means of the at least one coordinate measuring device 16, and the established coordinates of the ranges of the optical axes 2.1 and 4.1 are stored. The ranges of the optical axes 2.1 and 4.1 serve as reference axes.

[0061] Together with the sample carrier 7 and the sample 8, the carrier device 6 is controllably displaceable by means of the transport device 10. To this end, the transport device 10 has at least one drive 15 which is actuatable by control commands of the control unit 14. The carrier device 6 can be delivered by means of the transport device 10 into one of the optical axes 2.1 and 4.1. Here, the current coordinates of the carrier device 6 are continuously captured or can be captured by means of the coordinate measuring device 16.

[0062] The computing unit 13, the control unit 14, one drive 15 or a plurality of drives 15 (see, for example, Figure 4 ) and the at least one coordinate measuring device 16 are connected to one another in a manner suitable for the exchange of data (indicated in the figures).

[0063] The computing unit 13 already mentioned is also implemented to compare the current coordinates of the carrier device 6 captured by means of the coordinate measuring device 16 with the current coordinates of the optical axes 2.1, 4.1. In the illustrated case of a displacement of the carrier device 6 from the first optical axis 2.1 to the second optical axis 4.1, the comparison of the captured current coordinates of the carrier device 6 is effected with the coordinates of the second optical axis 4.1.

[0064] In particular, the comparison is between the coordinates of the reference point 11 set on the first optical axis 2.1 and the (positional) coordinates of the second optical axis. Depending on the difference established by means of the comparison between the current coordinates of the reference point 11 and the coordinates of the second optical axis 4.1, a control command is generated in the control unit 14 and sent to the at least one drive 15 which controls the delivery movement of the carrier device 6, which is thus actuated accordingly. The control is thus effected such that the carrier device 6 is positioned in such a way that the reference point 11 coincides with the second optical axis 4.1. The carrier device 6 is then located at the second measuring position P2 (indicated in an exemplary manner by a dashed line and marked by an arrow).

[0065] Figure 1b The same procedure is shown in a simplified manner in plan view in Fig. 4.

[0066] Figure 1a and 1bThe first exemplary embodiment shown in the middle has a transport device 10, which is common to the microscope 1 and the further microscope 3. The transport device 10 is embodied in exemplary manner as a guide rail system.

[0067] The two microscopes 1, 3 can be stored in different housings or housing parts from one another, indicated by the dashed line between the two microscopes 1, 3. This is advantageous, for example, if the microscopes 1 and 3 are intended to be operated at different environmental conditions.

[0068] Figure 1a and 1b The central idea and the advantages of the microscopy arrangement M according to the application are elucidated. The two microscopes 1 and 3 share a common transport device 10. This allows the sample 8 to be positioned relative to the optical axes 2.1, 4.1 at the respective measurement positions P1, P2 and to be transported between the measurement positions P1, P2. The sample 8 thus never leaves the reference coordinate system of its sample holder 7. The three-dimensional positioning of the sample 8, i.e. relative to the axes x, y and z, is facilitated, for example, by setting and continuously establishing the reference point 11.

[0069] As an example, only some possible embodiments of the first microscope 1 and the further microscope 3 are presented. As an example, the first microscope 1 and / or the further microscope 3 is embodied as an inverted light sheet microscope, as an upright light sheet microscope or as a microscope with very high resolution and, for example, only one objective. In particular in the latter embodiment, the offset can be reduced or even avoided. The first microscope 1 and / or the further microscope 3 can also be embodied, for example, to incubate the sample 8 under predetermined temperature conditions or regimes, gas conditions and / or illumination conditions or regimes. In addition, the first microscope 1 and / or the further microscope 3 can be embodied to capture overview images and / or as a microscope with an objective exchange device. In addition, the first microscope 1 and / or the further microscope 3 can be embodied as a scanning microscope, in particular a laser scanning microscope (LSM), as a microscope with a manipulation unit to manipulate the sample 8 by light beams, by acoustic elements and / or by mechanical elements.

[0070] The measurement positions P1, P2 are preferably optically independent of one another and are connected only by the common transport device 10. However, the illumination and / or detection beam paths can be partially superimposed by appropriate optical elements and thus the measurement positions P1, P2 both use light sources, fittings, modules, detectors or cameras.

[0071] Figure 2A second exemplary embodiment of a microscopy arrangement M is shown, wherein the elements of the microscopy arrangement M act upon both the illustrated measuring positions P1 and P2. In the exemplary embodiment, the detection beam paths of both microscopes 1 and 3 are superimposed by a semi-transparent mirror serving as a beam unifier 20 and directed onto a common detector 17, e.g., a camera. Depending on the distance between the measuring positions P1, P2, this can be achieved by two tube lenses T1, T2 with different focal lengths or by means of a relay optical unit (not shown). The beam unifier 20 and the tube lens T2 can be implemented as a group marked by a dashed edge. If the detection radiation from the measuring position P1 is to be imaged onto the detector 17, a relay optical unit (not shown) can be introduced into the detection beam path, e.g., instead of the group consisting of the tube lens T2 and the beam unifier 20.

[0072] Advantageously, the microscopy arrangement M facilitates a large number of options for combining the available microscopes 1, 3, ... n. Figure 3 As shown, a light sheet microscope is located at a first measurement position P1 of a third exemplary embodiment of a microscopy arrangement M according to the present invention. Light sheet 5 is generated in a region where sample 8 is located or can be located, using an illumination objective BO, which is designed to generate a light sheet 5 (illustrated in simplified form as a strip) along an illumination axis BO.1. Light sheet 5 can be generated dynamically by scanning a light beam in a plane, in this case at an angle of approximately 45° relative to sample plane 9. In other embodiments, light sheet 5 can be generated by an optical element (e.g., a cylindrical lens) arranged upstream of illumination objective BO. In simplified form, element 21 is shown as a representation of both options. First objective 2, also present in first microscope 1, serves as a detection objective. The optical axis of illumination objective BO extends at an angle of approximately 45° relative to sample plane 9. A first optical axis 2.1 provided by first objective 2 also extends at an angle of approximately 45° relative to sample plane 9 and is perpendicular to the optical axis of illumination objective BO (illumination axis BO.1) and intersects illumination axis BO.1 in the region of the generated light sheet LB.

[0073] The further microscope 3 has an objective exchange device (indicated by the designation of an additional objective). One of the objectives contained therein is in the working position for the second optical axis 4.1 and represents the further objective 4.

[0074] Figure 3 In the exemplary embodiment shown in FIG, a light sheet 5 is generated at a first measuring position P1 and the illuminated sample 8 is observed through a first objective 2. In addition, one or more correction elements 22 (e.g. suitably implemented meniscus lenses, Alvarez plates and / or relay optics) are provided to correct for a tilted passage through the sample carrier 7. Alternatively, the microscope 1 can be operated with special sample containers that avoid tilted passages.

[0075] In the exemplary embodiment shown, a special microscope 1, which does not allow or only allows in a restricted range conventional microscope method steps such as objective exchange, recording of overview images, alternating contrast (DIC, phase contrast, polarization contrast, etc.), is located at the measurement position P1. These method steps are facilitated at the measurement position P2. To this end, the further objective 4 is conventionally arranged, i.e. the optical axis 4.1 extends perpendicularly to the sample carrier 7 at the measurement position P2. In this exemplary embodiment, a common detector 17 in the form of a camera is used for both measurement positions P1, P2. The detection beam paths superimpose one another, and the distance of the measurement positions P1, P2 is chosen such that the sample planes 9 from the measurement positions P1, P2 are imaged by the tube lenses T1 and T2 with different focal lengths onto a common image plane (dashed line) on the detector 17.

[0076] If the sample 8 to be examined is at the measurement position P1, the group formed by the beam homogenizer 20 and the tube lens T2 is removed from the beam path, so that the detection beam path is introduced from the first objective 2 to the detector 17 without passing through this group.

[0077] If the sample 8 is examined at the measurement position P2, the group consisting of the beam homogenizer 20 and the tube lens T2 is introduced into the common beam path. The detection beam path of the objective 4 passes through said group and is diverted onto the detector 17.

[0078] In other embodiments, filter devices can also be used with both beam paths, which can be introduced into or removed from the respective beam path in a manual or motor-driven manner. For the common use of optical elements, additional common infinite spaces can be created.

[0079] The sample 8 is moved between the measurement positions P1 and P2 by the common transport device 10. In other embodiments of the microscopy arrangement M, there can be further measurement positions Pn (n = 3, 4,... n, not shown). In an advantageous configuration, the transport device 10 allows both a change in position in the x direction and a fine positioning of the sample 8 in the x, y and z directions at the respective measurement positions P1, P2. Thereby, all positioning tasks at the measurement positions P1, P2,... Pn can be carried out with a minimum degree of freedom of the actuators. This also includes the focusing (z direction), so that, for example, two special measurement positions P1, P2 can be connected to one another by the solution according to the application and a z drive of, for example, the focusing objectives 2, 4 or the objective exchange device can be omitted.

[0080] In Figure 3In the microscopy arrangement M, which is shown schematically, a transillumination illumination unit 23, which is composed of a light source 24 and a condenser 25, can be provided, by which the sample 8 can be illuminated from above. Either the illumination unit 23 can be provided at both measurement positions PI, P2, or it is coupled to the transport device 10 and moved therewith from the measurement position PI to the measurement position P2 and vice versa. Fluorescence (reflected light) illumination can be coupled in at the measurement position P2 by means of the dichroic beamsplitter 18.

[0081] In other embodiments of the application, a relay optical unit can also be used if the tube lenses T1, T2 with different focal lengths are not used and / or the distance bridged between the common beam paths is too great.

[0082] Figure 4 The setup of a further example of a microscope for light sheet microscopy is shown schematically. As explained with respect to Figure 3 The illumination objective BO and the first objective 2, which generate the light sheet 5, are present. The illumination radiation is formed in a controlled manner by means of the AOTF 19 (acousto-optic filter) and deflected by means of the scanner 21. To this end, the AOTF 19 is connected to the control unit 14, which in turn is connected to the computing unit 13. The illumination radiation deflected by means of the AOTF 19 is formed into the light sheet 5 by means of the illumination objective BO, wherein the extent of the light sheet 5 is essentially determined by the deflection of the scanner 21, in particular in the direction of the y-axis. In other embodiments, instead of the AOTF 19, for example a spatial light modulator (SLM), a micro-mirror array (digital micro-mirror device, DMD) or an adaptive mirror can also be used. As an example, a laser and / or a (laser) diode can be used as a light source for the illumination radiation.

[0083] A so-called relay optical unit arrangement is present as a correction element 22 between the illumination objective BO or the first objective 2 and the sample carrier 7, by the effect of which aberrations that arise when the illumination radiation or the detection radiation passes through the sample carrier 7 are reduced.

[0084] In a development of the aforementioned exemplary embodiments, the use of certain elements, for example the light source for the illumination radiation, can be provided to a plurality of microscopes 1, 3, and corresponding beam paths and / or switching and selection options can be present.

[0085] Based on Figure 2A method of carrying out an operation microscopy arrangement M according to the application is described. In step A, a three-dimensional reference coordinate system is set as a Cartesian coordinate system having the axes x, y and z. Other reference coordinate systems, such as polar coordinate systems, cylindrical coordinate systems, can also be set in other configurations of the method. The coordinates of the range of the optical axis 2.1 of the first microscope 2 and the coordinates of the range of the other optical axis 4.1 of the other microscope 3, or of the optical axis of the microscope to be used next, are known or established and stored in a memory of the computing unit 13, for example. The optical axes 2.1, 4.1... serve as reference axes.

[0086] In step B, the sample carrier 7 with the sample 8 is introduced onto the sample plane 9 and the first optical axis 2.1. A reference point 11 is then set which coincides with the first optical axis 2.1 and the coordinates of the reference point 11 are established in the reference coordinate system (step C). As an example, the reference point 11 lies in the region of the sample 8 to be examined (region of interest, ROI). Subsequently, the sample 8 can be analyzed using the first microscope 2 and the examination method or the examination method carried out by the first microscope 2. Once the analysis has been carried out, a signal is provided by the computing unit 13 to the control unit 14, which then causes the generation of at least one control command and the transmission of the control command to the drives 15 of the transport device 10, so that the drives 15 are actuated in accordance with the control command obtained and the sample holder 7 is transported together with the sample 8.

[0087] In the delivery of the carrier device 6 to the other microscope 3, the current coordinates of the reference point 11 are continuously captured and compared with the current coordinates of the optical axis 4.1 of the other microscope 3. For this purpose, the computing unit 13 is used. Depending on the difference established by the coordinate comparison between the current coordinates of the reference point 11 and the optical axis 4.1 of the other microscope 3, a control command is generated by the control unit 14 after the computing unit 13 has transmitted the appropriate information to the control unit 14. The generated control command is transmitted to at least one of the drives 15, and the other delivery movement of the carrier device corresponding to the respective control command is controlled. As a result of the continuous capturing of the current coordinates and the comparison thereof with the current data of the reference axis, in particular the second optical axis 4.1 specified in the example, and the control command generated as a result, the carrier device is positioned in such a way that the reference point 11 coincides with the other optical axis 4.1.

[0088] In other configurations, the reference point 11 is brought into correspondence with the target point 12 on the second optical axis 4.1, so that the sample 8 is also positioned in the direction of the z axis.

[0089] Reference signs

[0090] M microscopy arrangement

[0091] 1 first microscope

[0092] 2 first objective

[0093] 2.1 first optical axis

[0094] 3 further microscope

[0095] 4 further objective

[0096] 4.1 second optical axis

[0097] 5 light sheet

[0098] 6 carrier device

[0099] 7 sample carrier

[0100] 8 sample

[0101] 9 sample plane

[0102] 10 transport device

[0103] 11 reference point

[0104] 12 target point

[0105] 13 computing unit

[0106] 14 control unit

[0107] 15 drive

[0108] 16 coordinate measuring device / sensor

[0109] 17 detector

[0110] 18 dichroic beam splitter

[0111] 19 acousto-optic filter

[0112] 20 beam unifier

[0113] 21 scanner / cylindrical lens

[0114] 22 correction element

[0115] 23 illumination unit

[0116] 24 light source

[0117] 25 condenser

[0118] BO illumination objective

[0119] BO.1 illumination axis

[0120] T1 tube lens

[0121] T2 tube lens

[0122] P1 first measurement position

[0123] P2 second measurement position

Claims

1. A microscope arrangement comprising: a first microscope and at least one other microscope, wherein each of the microscopes has an optical axis, the optical axis of the first microscope not coinciding with the optical axis of the at least one other microscope; a carrier device embodied to receive and hold a sample carrier in the microscope arrangement; a coordinate measuring device by which current coordinates of the carrier device are continuously captured; a computer carried in the microscope arrangement to set up a three-dimensional reference coordinate system, wherein coordinates of the extent of the optical axis of the first microscope and coordinates of the extent of the optical axis of the at least one other microscope are known and stored, or wherein coordinates of the extent of the optical axis of the first microscope and coordinates of the extent of the other optical axis of the at least one other microscope established by at least one of the coordinate measuring devices are stored and taken as reference axes; a transport device embodied in the microscope arrangement to deliver the carrier device to one of the optical axes, while the transport device is embodied to transport the carrier device from a first measurement position, in which imaging using the first microscope takes place, to a second measurement position, in which imaging using a second microscope takes place; a common detector for receiving detection light from detection beam paths of the first and second microscopes, while the detection beam paths of the first and second microscopes are superimposed on one another by a beam unifier and introduced along a common beam path into a common image plane of the common detector, the computer is configured to compare the current coordinates of the carrier device captured by the coordinate measuring device with the coordinates of the optical axis of the at least one other microscope, wherein a comparison between the coordinates of a reference point set on the optical axis of the first microscope and the coordinates of the optical axis of the at least one other microscope is implemented; a controller generates a control command depending on a difference established by the comparison between the current coordinates of the reference point and the coordinates of the other optical axis and controls the delivery movement of the carrier device depending on the control command such that the carrier device is positioned in such a way that the reference point coincides with the other optical axis.

2. The microscope arrangement according to claim 1, wherein the first microscope and / or the at least one other microscope each have an illumination objective and a detection objective, whose respective optical axes are perpendicular to one another and are directed into a sample plane and contain an angle different from zero with the sample plane and a normal to the sample plane, a sample being arranged or being able to be arranged in the sample plane; the illumination objective is configured to produce a light sheet intersecting the sample plane.

3. The microscope arrangement according to claim 2, wherein the illumination objective and the detection objective are arranged in an inverted manner such that their optical axes are directed or are able to be directed through a sample carrier onto the sample plane.

4. The microscope arrangement according to claim 1, wherein if a detection beam from the image plane of the first measurement position is to be detected, the sample planes of the first and second measurement positions are imaged by a first tube lens or relay optical unit onto a common image plane on a common detector; or if a detection beam from the image plane of the second measurement position is to be detected, the sample planes of the first and second measurement positions are imaged by a second tube lens in the common beam path onto a common image plane on a common detector; wherein the first and second tube lenses have different focal lengths.

5. The microscope device according to claim 1, wherein the beam unifier and the second tube lens form a set which can be introduced into or removed from the common beam path, respectively.

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