Microscope system for imaging a sample area and corresponding method

Abstract

Microscope system (100) for imaging at least one area of ​​a sample with an image generation device (110) which is set up for microscopic imaging of a three-dimensional sub-area of ​​the sample area to be imaged, captured in an observation beam path (118), a moving device (120) which is configured to move the three-dimensional sub-area to be imaged into the observation beam path (118) of the image generation device (110), a graphical user interface (130) displayed on a display (131), which is configured to receive at least one user input prior to the actual imaging for defining a three-dimensional travel range (133) corresponding to the sample area to be imaged, wherein the graphical user interface is configured to display a three-dimensional coordinate system (132), wherein the selection of at least one point (A, B, C) is made by user input in the three-dimensional coordinate system (132) and after selection of this at least one point (A, B, C, D) in the displayed coordinate system (132) the three-dimensional travel range (133) is defined in the form of a travel volume, and a control device (140) which, depending on the defined travel volume (133), controls the travel device (120) in such a way that it moves a quantity of sub-areas corresponding to the defined travel volume successively into the observation beam path (118) and the sub-areas are each imaged by the image generation device (110).

Description

[0001] The present invention relates to a microscope system for imaging at least one region of a sample, comprising an image generation device for microscopically imaging a partial region of the sample region to be imaged, which partial region is located in a focus of an observation beam path, a displacement device or scanning device which is configured to move the partial region to be imaged into the focus of the observation beam path of the image generation device, and a control device which, depending on a displacement range (scanning range) to be defined, controls the displacement device in such a way that a predetermined number of partial regions are approached within the defined displacement range and imaged by the image generation device.

[0002] Such microscope systems are generally known from the prior art. They are used, for example, when a sample or the sample area to be imaged exceeds the dimensions that a microscopic image can capture in the field of view of the imaging optics. Often, an overview image of a portion of the sample can be generated at low magnification. A user then selects, for example, the sample area of ​​interest, which is then traversed or scanned according to a defined pattern. The sample area to be imaged is imaged by tracing and imaging a predetermined number of partial areas according to the predetermined scan pattern. The generated images are then superimposed using a suitable image processing program and combined to form an overall image (mosaic image) of the sample area.Typically, the sample area to be imaged is a sample volume extending in the x, y, and z directions. The sample area is usually scanned in several x and y planes at different z coordinates. For this purpose, the microscope system typically has a microscope stage that can be moved in the x and y directions, as well as a focusing device that can vary or fix the focus of the microscope system's observation beam path in the z direction. For focusing purposes, the objective of the microscope system and / or the aforementioned microscope stage can be moved in the z direction.

[0003] With such conventional microscope systems, defining the sample area to be imaged or the associated travel range (scan range) often proves to be problematic and not very user-friendly. For example, the scan range is currently defined using sliders that can be used to define the scan areas in the x, y, and z directions. If the scan range is set too large, the image will include areas that either do not contain a sample or contain sample areas of no interest. Both of these factors increase the effort and time required for image acquisition. If the scan ranges are set too small, there is a risk that the sample volume of interest will not be fully imaged.

[0004] PROCEEDINGS OF SPIE, "Acquisition of multiple image stacks with a confocal laser scanning microscope", W. Zuschratter et al., Image Acquisition and Processing V, (9 June 1998); doi: 10.1117 / 12.310551, describes a confocal microscope system from the applicant for acquiring so-called "multiple image stacks", i.e., a large number of composite images. Up to 70 positions in the xy plane can be saved and selected via a graphical user interface. A starting position can be set using visual control via the microscope. Additional positions in the xy direction are then set by clicking until the region of interest is registered. Thus, at least two clicks and a maximum of 70 clicks are required to define an area. To define the z-travel range, the sample is focused in the z-direction and the number of steps in the z-direction is then set using another menu ("memory control window").The z-range can be set separately for each image stack.

[0005] DE 100 58 100 A1 discloses a method for scanning microscopic objects with a scanning device. A marking device can be used to encircle an object area to be examined, thereby defining a boundary line within which a plurality of two-dimensional scan fields are automatically distributed. According to this document, a three-dimensional image can be obtained by layer-by-layer image data acquisition. A scan field is moved accordingly in the z-direction by a relative movement between the sample stage and the focusing optics.

[0006] DE 101 08 240 B4 describes a method for imaging and measuring microscopic three-dimensional structures. This method already requires the availability of three-dimensional image stacks acquired with a microscope. This document aims to facilitate measurement tasks, such as measuring lengths running diagonally within a volume. For this purpose, the length running within a volume is displayed to the user on a screen in a section along this length. The length can then be determined by marking the starting and end points on a PC.

[0007] The object of the present invention is to provide a way to define the travel range or scan range corresponding to the sample area to be imaged more precisely and in a more user-friendly manner. Disclosure of the invention

[0008] This problem is solved by the subject matter of the independent patent claims. Advantageous embodiments emerge from the respective subclaims and the following description.

[0009] The microscope system according to the invention is a microscope system of the type mentioned at the beginning. In order to image at least one area of ​​a sample, the microscope system has an image generation device for microscopically imaging a partial area of ​​the sample area to be imaged, which partial area lies in a focus of an observation beam path. Such an image generation device comprises, for example, at least one microscope objective, a magnification changer and / or a zoom system and generally a tube with a downstream camera. Tubeless microscopes are also known. For example, the microscope image can be imaged directly onto an image sensor. In this way, a partial area lying in the observation beam path can be microscopically imaged in a known manner. Furthermore, the microscope system has a traversing orA scanning device configured to move the respective partial area to be imaged into the observation beam path of said image generation device. For this purpose, the microscope stage supporting the sample is moved and / or the focusing device of the microscope system is adjusted accordingly. For focusing purposes, the objective lens and / or the microscope stage can be moved in the focus direction. In this way, a partial area to be imaged can be approached in all three spatial directions and imaged by the image generation device. The partial areas moved into the observation beam path are advantageously arranged at a focus of the image generation device.

[0010] The microscope system according to the invention further comprises a graphical user interface which is shown on a display and serves to define a travel range or scan range corresponding to the sample area to be imaged by at least one user input.

[0011] A "graphical user interface" can be understood as a graphical display generated by a program. On this display, a user can enter instructions into the microscope system. The instructions are converted into actions by the microscope system. The user inputs data directly into the display, for example, by moving a mouse pointer (cursor) and clicking or selecting desired positions. If the display is a touch-sensitive or capacitive screen (touchscreen), input can be made using a finger or a stylus suitable for the screen.

[0012] The graphical user interface is configured to display a coordinate system, and by entering at least one point in the coordinate system, the travel range is defined in the form of a travel volume. Depending on the defined travel volume, the control device of the microscope system controls the travel device such that, within the defined travel volume, a predetermined number of subregions are moved to and imaged by the image generation device. In other words, a number of subregions corresponding to the travel volume are moved one after the other into the observation beam path, and the subregions are each imaged. The images of the subregions generated in this way are superimposed and combined in a known manner to obtain an image of the sample region to be imaged.The number and positions of the sub-areas within the volume depend on the required image quality of this image.

[0013] The graphical user interface is at least configured in such a way that a user can specify or define the travel range corresponding to imaging the desired sample area. For this purpose, a coordinate system is displayed on the graphical user interface, within which the user can define a travel or scan volume by entering one or more points. The travel range defined in this way is displayed by the user interface, for example in the form of a clearly defined travel volume. After confirmation of the defined travel volume, the scan then takes place, for example, with the respective image generation. For this purpose, the graphical user interface can, for example, be connected to a computing unit designed to generate signals that are passed to the control device.Such a computing unit can also be present separately, for example in the display or on a graphics card.

[0014] It is particularly advantageous if at least one, but especially every, of the points entered by the user lies on or within the sample area to be imaged. It is particularly useful if a user defines three points in succession that represent the boundary points of the sample area to be imaged. By selecting at least three such points in space, a corresponding sample volume can be selected, and the corresponding travel range can be defined as the travel volume and displayed using the graphical user interface. Scanning then takes place within the defined travel volume, generating the respective image.

[0015] Without loss of generality, it should be assumed in the following that the displayed coordinate system is a Cartesian coordinate system in the x, y, and z directions. By selecting at least three points in such a coordinate system – provided at least one of these points is not in the same plane as the other points – a three-dimensional travel volume can be defined. Again, without loss of generality, a cuboid-shaped travel volume should be assumed. Of course, other travel volumes can also be defined in a Cartesian coordinate system by specifying at least three points. Furthermore, another coordinate system can also be used, such as a spherical or cylindrical coordinate system. When x, y, and z coordinates are mentioned in the following, this is not intended to limit the general inventive concept. The same applies to the shape of the defined travel volumes.

[0016] A first advantageous possibility for defining a cuboid-shaped travel range is described below.

[0017] The graphical user interface first enters a first point A and a second point B in a common xy plane in the displayed coordinate system. Point A is advantageously the current focus position, with point A forming an edge point or limit value of the travel range to be defined. The sample is then moved further by moving the microscope stage in the xy plane until it reaches a second limit value B, which the user confirms, for example, by a suitable input via a button or with a mouse click. The graphical user interface is set up in such a way that a predefined rectangular travel range is defined, with points A and B forming edge points or limit values ​​of this travel range. It is particularly expedient if points A and B form opposite corners of the rectangular travel range.Based on the definition of the rectangular travel range, an image scan can in principle already be performed, although this is then limited to the selected xy plane. To define a travel volume, the user enters a third point C in the same displayed coordinate system, which lies outside the xy plane passing through points A and B. Advantageously, the graphical user interface is set up such that the entered third point C forms an edge point of the travel range, in particular a cuboid-shaped one. Point C lies, for example, in a plane that is opposite the plane spanned by points A and B and runs parallel to it. Point C advantageously lies on or in the sample area to be imaged. Again, it can be expedient if point C lies at a corner of the cuboid-shaped travel range.For example, starting from one of the points A or B, the user can move the sample area in the z-direction to reach a point that he defines as point C. Starting from the three specified points A, B and C, the graphical user interface defines the cuboid-shaped travel volume.

[0018] After defining the cuboid-shaped travel volume, the microscope system's control unit controls the travel unit in a predetermined manner so that the defined sample area is scanned. For example, images of partial areas are first acquired in an xy plane, then in a plane parallel to this xy plane, and then continuously until the entire scan volume has been scanned. An image is generated at each scan position, and the images acquired at the various scan positions are superimposed and combined to form an overall image of the sample area.

[0019] It will be apparent to those skilled in the art that other predefined travel volumes can also be defined by entering three points in the space of a coordinate system. In principle, almost any geometric body can be specified by defining three boundary points of this body. In practice, with microscope systems with microscope stages that can be moved in the xy direction and focusing devices that can be moved in the z direction, a cuboid-shaped travel volume is generally most suitable.

[0020] Furthermore, it is not absolutely necessary that the three specified points A, B and C all lie in or on the sample area to be imaged.

[0021] For example, in the above example of defining a cuboid-shaped travel range, point C can be set at a height (z-coordinate) that ensures that all z-coordinates of the sample area to be imaged are captured. For this purpose, point C can be set slightly higher than the actual sample length. The same considerations apply in a completely analogous manner to points A and B.

[0022] In the following, another advantageous possibility of defining a cuboid-shaped travel range is described.

[0023] With this additional option, it is sufficient to enter only two points A and C into the coordinate system displayed by the graphical user interface. For this purpose, the graphical user interface is configured to treat the two entered points as edge points of a cuboid-shaped travel area. Advantageously, the two points A and C define opposite corner points of the cuboid-shaped travel volume; more precisely, they form the starting and end points of a spatial diagonal of the cuboid-shaped travel volume. Further explanations of this configuration can be found in the exemplary embodiments at the end of the description.

[0024] Finally, another advantageous option for defining a cuboid travel range or, more generally, any predefined travel volume should be noted. For this purpose, the graphical user interface can be configured such that, starting from a point A entered in the displayed coordinate system, a predefined volume outline is displayed in the coordinate system, by means of which the travel volume is defined or which defines the travel volume. For example, a cuboid volume outline can be displayed in the coordinate system, at one corner of which the entered point A is located. Such a configuration is useful, for example, when sample areas of the same or similar dimensions are to be examined in chronological succession, so that the travel volume changes little.In this case, specifying a single corner or edge point (point A) of this travel volume may be sufficient to define it.

[0025] Furthermore, the graphical user interface can be configured such that a surface or spatial grid consisting of the approach points corresponding to the sub-areas to be mapped is displayed in the coordinate system containing the traversing volume. For example, a cuboid-shaped spatial grid can be displayed in the coordinate system, with point A, for example, located at one of its corners. The approach points corresponding to the sub-areas are marked in the spatial grid.

[0026] In general, it is advantageous if the defined travel volume is displayed in the coordinate system, for example, in the form of a volume outline or spatial grid that illustrates the limits of the travel range. It can also be useful if the maximum possible travel volume or a maximum travel range is displayed in the coordinate system. The maximum possible travel volume is determined by the system parameters of the microscope stage and the focusing device, i.e., by the respective maximum travel distances in the x, y, and z directions. With this configuration, the user can see how far they are from the limits of the maximum possible travel volume.

[0027] To locate the sample area of ​​interest, images of sub-areas of the sample are often taken. These images may have been taken previously or during the definition of the traversing volume. Such images, or even acquired image sequences in the z-direction, can be displayed by the graphical user interface in a separate display area or—for example, semi-transparently—in the coordinate system to facilitate the user's definition of the traversing volume.

[0028] In an advantageous embodiment, the graphical user interface is configured such that, after the travel volume has been defined, it can be modified or redefined by entering at least one additional point. For example, by selecting and entering an additional point D that lies outside the defined travel volume, the travel volume can be expanded, while by entering an additional point D that lies within a defined travel volume, this travel volume can be reduced. In this case, it is particularly expedient if the additional point D entered forms a new boundary point of the newly defined, in particular expanded or reduced, travel volume. This new boundary point can in particular be a new edge or corner point, for example of a cuboid-shaped travel area.In this embodiment, it may also be expedient if the graphical user interface is configured such that by selecting or clicking an edge point of the displayed defined travel volume, this can be expanded or reduced in a defined manner, in particular while maintaining the cuboid shape. Once the travel volume has been modified as desired, the user confirms this by entering a value (e.g., releasing the clicked edge point) at the new point D.

[0029] The invention further relates to a method for imaging at least one region of a sample by means of a microscope system, wherein a partial region of the sample region to be imaged, which lies in an observation beam path of an image generation device of the microscope system, is imaged by means of the image generation device, the partial region to be imaged is moved into the observation beam path of the image generation device by means of a movement device of the microscope system, a movement region corresponding to the sample region to be imaged is defined by at least one user input in a graphical user interface of the microscope system, wherein the user interface displays a coordinate system and the movement region is defined in the form of a movement volume by entering at least one point (A, B, C, D) in the coordinate system,and wherein the displacement device is controlled by a control device of the microscope system depending on the defined displacement range such that a predetermined number of partial areas are moved to within the defined displacement range and imaged by the image generation device.

[0030] With regard to the method according to the invention, its embodiments and advantages, reference is expressly made to the explanations of the microscope system according to the invention.

[0031] According to an advantageous embodiment, at least one of the at least one point (A, B, C, D) to be input is selected such that it lies on or in the sample area to be imaged.

[0032] According to an advantageous embodiment, the travel volume is defined by the graphical user interface by entering at least three points (A, B, C), wherein the three points (A, B, C) are used as boundary points of the travel volume.

[0033] According to a further advantageous embodiment, a Cartesian xyz coordinate system is used as the displayed coordinate system.

[0034] In a further advantageous embodiment, the graphical user interface initially defines a rectangular travel range as the travel range if a first and a second point (A, B) in a common plane are entered in the coordinate system displayed by the graphical user interface, and subsequently defines a cuboid travel volume as the travel range if a third point (C) outside the plane passing through the first and second points (A, B) is entered in the same displayed coordinate system. The plane selected initially can be the xy plane, which lies parallel to the stage surface of the microscope stage. However, the xz plane or the yz plane, which are each perpendicular to the stage surface of the microscope stage, can also be selected as the starting surface. The third spatial direction is then selected using the third point (C).

[0035] In a further advantageous alternative embodiment, the graphical user interface uses the input first, second, and / or third point (A, B, C) as boundary points of the cuboid travel volume. In a further advantageous embodiment, the graphical user interface defines a cuboid travel volume as the travel area by entering a first and a second point (A, C) in the displayed coordinate system, wherein the first and / or the second point are used as boundary points of the cuboid travel volume, in particular as the start and end points of a spatial diagonal of this cuboid volume.

[0036] In a further advantageous alternative embodiment, the graphical user interface displays a surface or spatial grid in the coordinate system from the approach points corresponding to the partial areas to be imaged, in which the travel volume is contained.

[0037] In general, it is useful if the graphical user interface displays the defined travel volume in the coordinate system.

[0038] In a further advantageous embodiment, the graphical user interface displays the maximum possible travel volume in the coordinate system.

[0039] In a further advantageous embodiment, the graphical user interface displays previously imaged sample regions. These previously imaged sample regions can serve as a reference point for the user when entering the selection points for the desired travel volume. For example, the previously imaged sample regions can be displayed as semi-transparent planes or volumes in the coordinate system.

[0040] In a further advantageous embodiment, after the first travel volume has been defined, a changed travel volume is redefined or modified in the graphical user interface by entering at least one further point (D).

[0041] In a further advantageous embodiment, the graphical user interface uses the at least one further input point (D) as a new boundary point of the newly defined travel volume.

[0042] Finally, the invention relates to a computer program with program code, wherein, when executed on a computing unit of a microscope system according to the invention, a method according to the invention is carried out. Furthermore, the invention relates to a corresponding computer program product having a computer program with program code stored thereon, wherein, when executed on a computing unit of a microscope system according to the invention, a method according to the invention is carried out. The computer program can be downloaded or uploaded as such, or stored or temporarily stored on a computer program product. Volatile or non-volatile storage media, such as USB sticks, RAM or ROM memories of known types, can be considered as the computer program product. The aforementioned computing unit of a microscope system according to the invention can be the control device or a part of this control device of the microscope system.

[0043] Further advantages and embodiments of the invention will become apparent from the description and the accompanying drawings.

[0044] It is understood that the features mentioned above and those to be explained below can be used not only in the combination specified in each case, but also in other combinations or on their own, without departing from the scope of the present invention.

[0045] The invention is illustrated schematically in the drawing using an embodiment and is described below with reference to the drawing. Character description Fig. 1 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in a first view. Fig. 2 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in a further view. Fig. 3 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in a further view. Fig. 4 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in a further view. Fig. 5 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in a further view. Fig. 6 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in an alternative embodiment. Fig. 7 schematically shows at least a section of a graphical user interface of a microscope system according to the invention in another alternative embodiment. Fig. 8 schematically shows an embodiment of a microscope system according to the invention. Fig. 9 schematically shows another embodiment of a microscope system according to the invention.

[0046] The figures are described comprehensively, with the same reference numerals designating the same elements. Fig. 1 to 5, an embodiment of a graphical user interface 130 for defining a travel range or scan range corresponding to a sample area to be imaged using user inputs to be explained will be described. The graphical user interface 130 is shown on a display 131 or monitor or display system, with a coordinate system 132 being displayed on the display 131 in the graphical user interface 130. In the present embodiment, this is a Cartesian coordinate system with x, y, and z axes. 135 denotes the maximum possible travel range, as specified by the maximum and minimum positions of an xy microscope stage of the microscope system, which is displaceable in the x and y directions.

[0047] Fig. 2 shows the graphical user interface 130 after a user has entered a point A. This point A is the current focus position; the corresponding image of the sample portion associated with point A is displayed, for example, in another (not shown) area of ​​the graphical user interface 130 or on another (not shown) display area of ​​the display 131 or on a separate display (not shown).

[0048] In another, in Fig. In the step shown in Figure 3, a user enters a second point B in the same xy plane. Advantageously (but not necessarily), the sample region corresponding to point B is moved into the focus of the observation beam path of the microscope system's image generation device, and a corresponding image is generated. In this way, the user can check whether point B is still in the sample region of interest or still within the sample. The coordinates of points A and B can be described as follows: A = (x A , y A , z1); B = (x B , y B, z1). After confirmation by the user, a rectangular travel area 134 is first defined in the graphical user interface 130, with points A and B in this embodiment forming opposite corner points of the rectangular travel area 134. The rectangular travel area 134 extends in the x-direction between the coordinates x A and x B and in the y-direction between the coordinates y B to y A .

[0049] In a next, in Fig. In the step shown in Figure 4, the scan area is defined in the form of a travel volume 133 by entering another point C into the same coordinate system 132 shown. For this purpose, only a point C is to be entered that lies outside the xy-plane passing through points A and B. This point C can be represented by its coordinates as C = (x C , y C , e.g. C). The graphical user interface 130 is configured such that parallel to the rectangular travel area 134, an xy-plane at the height of the coordinate z C from which a cuboid-shaped travel volume 133 can be defined as the scan volume. In the embodiment shown according to Fig. 4, point C lies on an edge of this cuboid traversing volume 133.

[0050] Fig. 5 finally shows a possibility of modifying the travel volume 133. Here, by entering a further point D, which is outside the Fig. 4 first defined travel volume 133, a new, modified travel volume 133' is defined. The point D can be defined by the coordinates (x D , y D , e.g. D). The resulting extended travel volume 133' is defined in the graphical user interface 130 such that the travel volume extends from the smallest x-value of the entered points to the largest x-value of the entered points, from the smallest y-value of the entered points to the largest y-value of the entered points, and finally from the smallest z-value of the entered points to the largest z-value of the entered points. In the embodiment according to Fig. 5 the travel volume 133' extends in the x-direction from x D up to x B , in y-direction of y B to y D and in the z-direction from z1 to z D .

[0051] When selecting points C and / or D, the corresponding sub-area of ​​the sample can be conveniently moved into the focus of the observation beam path of the microscope system's imaging device in order to display corresponding images of the sub-areas to the user. This makes it easy to check, for example, whether point D still corresponds to a point within the sample area of ​​interest to be imaged or within the sample.

[0052] After defining the travel volume 133 or 133' via the graphical user interface 130, the sample scan begins, for example, at coordinate z1. Then, the corresponding xy-rectangular area is scanned, for example, in lines or in a meandering pattern. Images of the corresponding sub-areas of the sample are acquired at predefined coordinates. Subsequently, scanning is performed in the same way in a parallel scan plane at a different z-coordinate, and images are acquired, and so on, until a final rectangular scan area at coordinate z Dis reached. Once this final scan plane has been scanned, the acquired images can be processed. Typically, the images from an xy plane are combined into a mosaic image, resulting in an image of the corresponding sample area in that plane. This creates a z-stack, or image stack, of images that can then be processed into a three-dimensional image of the corresponding sample volume.

[0053] Fig. Figure 6 shows another alternative for defining a cuboid travel volume, where a user enters a first point A and a second point C in the coordinate system 132 displayed in the graphical user interface 130. Points A and C form the edge points of the cuboid travel volume, here—for a better understanding of the underlying principle—two opposite corner points of the cuboid travel volume 133. Points A and C thus form the two endpoints of the spatial diagonal of the cuboid, which defines the travel volume 133. The two points A and C are defined analogously to the procedure already described above. It is only necessary to ensure that points A and C do not lie in the same xy plane, since otherwise only a two-dimensional, rectangular travel area would be defined. Again, it is expedient if points A and C lie in or on the sample area to be imaged.

[0054] After entering the two points A and C, a computing unit of the microscope system 100 calculates the cuboidal travel volume 133" and displays it in the graphical user interface. The computing unit can also be at least partially a component of the user interface or separately, for example, in the display device or a graphics card, or finally, can itself be a component of a more comprehensive computing unit that is arranged in the control device 140 or another component of the microscope system 100. The coordinates of the points A and C can be denoted by (x A , y A , e.g. A ) or (x C , y C , e.g. C ). The cuboid-shaped travel range 133" extends in the x-direction from x A up to x C , in y-direction of y C to y A and in the z-direction of z A to z C .

[0055] After defining the cuboidal travel range 133", the scan with image generation takes place in the same way as already described above using the Fig. 1 to 5 described.

[0056] Fig. 7 again shows an alternative possibility for defining a travel range 133''' corresponding to the sample area to be imaged, wherein in this case the graphical user interface 130 is configured such that by entering a single point A by a user into a coordinate system 132 displayed on the graphical user interface 130, a spatial grid or volume outline 136 is displayed starting from the point A in the coordinate system 132, by means of which the travel volume 133''' is defined.

[0057] In the Fig. 7, a cuboidal travel volume 133''' of predetermined dimensions is generated at a predetermined location, here starting from point A, and is shown in the graphical user interface 130 on the display 131. The user can now confirm the suggested spatial grid or volume outline 136 by entering it, so that this spatial grid 136 determines the cuboidal travel volume 133'''. It can also be provided that a user, in an analogous procedure to the embodiment according to Fig. 5 can make a modification of the spatial grid 136. This can be done by entering another point (corresponding to point D in Fig. 5) that lies outside or inside the spatial grid 136 and is defined as the new edge or corner point of a new traversing volume. Alternatively, the spatial grid 136 can be clicked at a specific location, for example, at a corner point, and enlarged or reduced using the familiar mouse control by dragging.

[0058] Alternatively or additionally, the displayed volume outline 136 can also be used or understood as a spatial grid. In this case, the approach positions corresponding to the subregions to be imaged can advantageously be displayed in the spatial grid 136 as grid points (not shown). This provides the user with a quick overview of the arrangement of the subregions used to image the sample area.

[0059] In a comprehensive, advantageous embodiment, after entering at least one point A, B, C, or D in the coordinate system 132, the entered point is graphically highlighted. This can be, for example, colored or by a thicker or flashing dot.

[0060] After defining a suitable travel volume 133''', the scan with image acquisition is carried out analogously to the embodiments described above.

[0061] The Fig. 8 to 9 show examples of specific embodiments of a microscope system 100. The microscope system 100 is shown in a very simplified manner, since components known per se from the prior art will not be explained in detail here. The microscope system 100 has an image generation device 110, a movement device 120, a control device 140, and a graphical user interface 130. The control device 140 can include a computing unit. However, the computing unit can also be arranged in another component of the microscope system 100. The image generation device 110 is in Fig. 8 using the components microscope objective 112 and camera 111. Illumination is provided by a gooseneck light 114 and a ring light 113 guided annularly around the objective 112. The gooseneck light 114 and the ring light 113 can be activated individually or in combination. This allows illumination from any direction and at any angle. This is advantageous for samples and observed objects with a pronounced topology, allowing for a clearer image of the structures of the sample or observed object.

[0062] In this embodiment, the traversing device 120 essentially comprises an xyz microscope stage that can be moved in all three spatial directions. Thus, a displacement of the objective 112 in the z-direction is unnecessary.

[0063] The microscope stage 121 is controlled to move the sample in the xy plane and to focus on a partial area of ​​the sample by moving it in the z direction. The sample itself is located on a sample holder 122 on the microscope stage 121. The microscope stage 121 is controlled by a control device 140. As shown, the control device 140 also controls other components, such as the objective lens 112, ring light 113, camera 111, and gooseneck light 114. In this way, the desired parameters of the illumination and the camera can be suitably adjusted. The definition of the scan area and the subsequent scan with image generation are also controlled via the control device 140 together with the graphical user interface 130. In this exemplary embodiment, the control device 140 is wirelessly connected to the graphical user interface 130, with a WLAN connection 141 (Wi-Fi) being suitably used.Alternatively, a cable connection is of course also possible. In principle, the Wi-Fi connection enables a spatial separation of the graphical user interface 130 from the other components of the microscope system 100. For this purpose, it is displayed, for example, on a display 131 of a tablet computer that is connected to the microscope system 100 via Wi-Fi.

[0064] The display 131 shows the graphical user interface 130 analogous to the embodiments discussed above, wherein a coordinate system for defining a travel volume is displayed. On the display 131, for example, displays according to the Fig. 1 to 7. As shown in Fig. As can be seen in Figure 8, the display 131 shows, in addition to the graphical user interface 130, another larger display area 137, on which the partial area currently in focus of the image generation device 110 is expediently displayed. This makes it easier for a user to navigate in the sample and thus to identify the sample area to be imaged. In this way, a user can, for example, according to the embodiment of the Fig. 1 to 4, identify a sample region of interest and define a corresponding travel volume 133. After defining the travel volume 133 in the graphical user interface 130, the control device 140 controls the scan by controlling the travel device 120 and the synchronized image generation by controlling the image generation device 110. After scanning an xy plane, the processed image is displayed, for example, in the display area 137. The same applies to further xy planes until the entire defined z range has been scanned. A three-dimensional image of the imaged sample region can then be calculated, whereby a corresponding 3D representation can again be displayed in the display area 137.

[0065] Fig. 9 schematically shows a further embodiment of a microscope system 100 with the option of incident and transmitted light illumination. Again, the known components of such a microscope system will only be mentioned cursorily. The microscope system 100 has an image generation device 110, a movement device 120, a control device 140, and a graphical user interface 130. The image generation device 110 comprises, as essential components, a microscope objective 115 movable in the z-direction, as well as deflection elements and lenses (not further designated), and a camera 111 arranged in the observation beam path 118. The incident light illumination device arranged on the objective side is designated 116. Its light is directed to the objective 115 via a semi-transparent beam splitter.

[0066] The generated illumination beam path is directed to the sample via unlabeled deflection elements and the objective lens 115. Light emanating from the focus of a partial area of ​​the sample is conversely directed by the objective lens 115 and further unlabeled optical elements in the form of the observation beam path 118 to the camera 111, where an image of the partial area is generated. The sample itself is located on a sample holder 122 on an xy microscope stage 123. The xy adjustment of the microscope stage 123 and the focusing drive of the objective lens 115 in the z direction serve as the traversing device 120 (see the double arrows with the x, y, z directions).

[0067] Also shown is a transmitted-light illumination device 117, the light of which passes through the sample and then reaches the objective 115. The transmitted-light illumination device 117 has a further lens (not shown in more detail) for generating a transmitted-light illumination beam path that illuminates the sample on the sample holder 122. Transmitted light is received by the objective 115 and guided to the camera 111 in the form of an observation beam path 118. In this way, a sample can be observed and imaged using the microscope system 100 shown here in reflected-light and / or transmitted-light illumination. The corresponding image generation is controlled via the control device 140, which controls (at least) the components of the transmitted-light illumination device 117, microscope stage 123, camera 111, reflected-light illumination device 116, and the focusing drive of the objective 115. Furthermore, the control device 140 is connected to the display 131 orThe graphical user interface 130 is connected via a cable in the illustrated case. Of course, a Wi-Fi connection is also possible here as an alternative, as described in connection with . Fig. 8 was explained.

[0068] The graphical user interface 130 also displays a coordinate system 132 for defining a travel volume. The corresponding display and the corresponding procedure for defining such a travel volume can be similar to the embodiments according to the Fig. 1 to 7. The display 131 has, in addition to the graphical user interface 130, a larger display area 137 on which the images generated can be shown. For example, images of partial areas can be displayed here when identifying the sample area of ​​interest, which facilitates the selection of suitable points for defining a travel volume. After defining a travel area, the Fig.8, a scan with image generation is performed. For this purpose, for example, with the focus position of the objective 115 fixed, the microscope stage 123 is moved in the xy direction according to a predetermined pattern until an image of an xy plane of the sample region to be imaged is obtained. This image can then be displayed in the display area 137. Subsequently, another xy scan is performed with a changed z position of the objective 115. This process is repeated until the entire z range of the travel volume has been processed. From the resulting image stack (z-stack), a three-dimensional sample image can be calculated, which is then displayed, for example, as a 3D image in the display area 137. List of reference symbols 100 microscope system 110 Image generating device 111 Camera 112 lens 113 Ring light 114 Gooseneck light 115 lens 116 Incident illumination device 117 Transmitted light illumination device 118 Observation beam path 120 Traversing device 121 xyz microscope table 122 sample holders 123 xy microscope table 130 graphical user interface 131 Display 132 Coordinate system 133, 133', 133'', 133''' travel volume, travel range 134 rectangular travel range 135 maximum travel range 136 spatial grids 137 display areas 140 Control device 141 Wi-Fi connection A, B, C, D point

Claims

[1] Microscope system (100) for imaging at least one region of a sample with an image generation device (110) which is designed for microscopically imaging a three-dimensional partial region of the sample region to be imaged, which is captured in an observation beam path (118), a displacement device (120) which is designed to move the three-dimensional partial area to be imaged into the observation beam path (118) of the image generation device (110), a graphical user interface (130) shown on a display (131) which is configured to receive at least one user input prior to the actual imaging for defining a three-dimensional travel range (133) corresponding to the sample area to be imaged, wherein the graphical user interface is configured for this purpose in such a way that it displays a three-dimensional coordinate system (132), wherein the selection of at least one point (A, B, C) is carried out by user input in the three-dimensional coordinate system (132) and after selection of this at least one point (A, B, C, D) in the displayed coordinate system (132), the three-dimensional travel range (133) is defined in the form of a travel volume, and a control device (140) which, depending on the defined travel volume (133), controls the travel device (120) in such a way that it moves a number of partial areas corresponding to the defined travel volume one after the other into the observation beam path (118) and the partial areas are each imaged by the image generation device (110). [2] Microscope system according to claim 1, wherein at least one of the at least one point (A, B, C, D) to be input lies on or in the sample area to be imaged. [3] Microscope system according to claim 1 or 2, wherein the graphical user interface (130) is configured such that the travel volume (133) is defined by the input of at least three points (A, B, C), wherein the three points (A, B, C) form edge points of the travel volume. [4] Microscope system according to one of claims 1 to 3, wherein the graphical user interface (130) is configured such that the displayed coordinate system (132) is a Cartesian xyz coordinate system. [5] Microscope system according to one of claims 1 to 4, wherein the graphical user interface (130) is set up in such a way that a rectangular travel range (134) is first defined as the travel range when a first and a second point (A, B) in a common plane are entered in the coordinate system (132) displayed in the graphical user interface (130), and then a cuboid travel volume (133) is defined as the travel range when a third point (C) outside the plane passing through the first and second point (A, B) is entered in the same coordinate system (132) displayed in the graphical user interface (130). [6] Microscope system according to claim 5, wherein the graphical user interface (130) is configured such that the input first, second and / or third point (A, B, C) form edge points of the cuboid travel volume (133). [7] Microscope system according to one of claims 1 to 4, wherein the graphical user interface (130) is set up in such a way that a cuboidal travel volume (133'') is defined as the travel range when a first and a second point (A, C) are entered in the coordinate system (132) displayed in the graphical user interface (130), wherein the definition of the cuboidal travel volume is carried out in such a way that the first and / or the second point form edge points of the cuboidal travel volume or form the starting point and the end point of a spatial diagonal of the cuboidal travel volume. [8] Microscope system according to one of the preceding claims, wherein the graphical user interface (130) is configured such that the defined travel volume (133) is displayed in the coordinate system. [9] Microscope system according to claim 8, wherein the graphical user interface (130) is configured such that a surface or spatial grid (136) of the approach points corresponding to the partial areas to be imaged is displayed in the coordinate system in which the travel volume (133''') is contained. [10] Microscope system according to one of the preceding claims, wherein the graphical user interface (130) is configured such that a maximum possible travel range (135) is displayed in the coordinate system. [11] Microscope system according to one of the preceding claims, wherein the graphical user interface (130) is configured such that already imaged sample or partial areas are displayed before or during the definition of the travel volume. [12] Microscope system according to one of the preceding claims, wherein the graphical user interface (130) is configured such that, after a previously defined travel volume (133), a modified travel volume (133') is redefined by entering at least one further point (D). [13] Microscope system according to claim 12, wherein the graphical user interface (130) is configured such that the at least one further input point (D) forms a new edge point of the changed travel volume (133'). [14] Microscope system according to one of the preceding claims, wherein the partial regions moved into the observation beam path (118) are each arranged in a focus of the image generation device (110). [15] Method for imaging at least one region of a sample by means of a microscope system (100), wherein a three-dimensional partial area of ​​the sample area to be imaged, located in an observation beam path (118) of an image generation device (110) of the microscope system, is imaged by means of the image generation device, the three-dimensional partial area to be imaged is moved into the observation beam path of the image generation device by means of a moving device (120) of the microscope system, a three-dimensional travel range (133) corresponding to the sample area to be imaged is defined before the actual imaging by at least one user input in a graphical user interface (130) of the microscope system, wherein the graphical user interface displays a three-dimensional coordinate system (132) for this purpose, wherein the selection of at least one point (A, B, C) is carried out by user input in the three-dimensional coordinate system (132) and after selection of this at least one point (A, B, C, D) in the displayed coordinate system (132), the three-dimensional travel range (133) is defined in the form of a travel volume, and wherein the movement device is controlled by a control device (140) of the microscope system depending on the defined movement volume such that a predetermined number of partial areas are moved to within the defined movement volume and imaged by the image generation device. [16] Method according to claim 15, wherein at least one point (A, B, C, D) to be input is selected such that it lies on or in the sample area to be imaged. [17] Method according to claim 15 or 16, wherein the travel volume is defined by the graphical user interface by entering at least three points (A, B, C), the three points (A, B, C) being used as boundary points of the travel volume. [18] Method according to one of claims 15 to 17, wherein a Cartesian xyz coordinate system is used as the displayed coordinate system. [19] Method according to one of claims 15 to 18, wherein the graphical user interface first defines a rectangular travel range as the travel range when a first and a second point (A, B) are entered in a common plane in the coordinate system displayed by the graphical user interface, and then a cuboid travel volume is defined as the travel range when a third point (C) outside the plane passing through the first and second point (A, B) is entered in the same coordinate system displayed in the graphical user interface. [20] Method according to claim 19, wherein the graphical user interface assigns the input first, second and / or third point (A, B, C) as edge points of the cuboid travel volume. [21] Method according to one of claims 15 to 18, wherein a cuboidal travel volume is defined as the travel area with the graphical user interface when a first and a second point (A, C) are entered in the displayed coordinate system, wherein the first and / or the second point are used as edge points of the cuboidal travel volume or form the starting point and the end point of a spatial diagonal of the cuboidal travel volume. [22] Method according to one of claims 15 to 21, wherein the graphical user interface displays the defined travel volume in the coordinate system. [23] Method according to claim 22, wherein the graphical user interface displays a surface or spatial grid of the approach points corresponding to the partial areas to be imaged in the coordinate system in which the travel volume (133''') is contained. [24] Method according to one of claims 15 to 23, wherein the graphical user interface displays the maximum possible travel volume in the coordinate system. [25] Method according to one of claims 15 to 24, wherein the graphical user interface displays sample or partial areas already imaged before or during the definition of the travel volume in the coordinate system. [26] Method according to one of claims 15 to 25, wherein, after definition of a first travel volume (133), a modified travel volume (133') is newly defined by input of at least one further point (D) using the graphical user interface. [27] Method according to claim 26, wherein the graphical user interface detects the at least one further input point (D) as a new edge point of the changed travel volume (133'). [28] Method according to one of claims 15 to 27, wherein the partial regions moved into the observation beam path (118) are each arranged in a focus of the image generating device (110). [29] Computer program with program code, wherein when executed on a computing unit of a microscope system (100) according to one of claims 1 to 14, a method according to one of claims 15 to 28 is carried out. [30] Computer program product having a computer program with program code stored thereon, during the execution of which on a computing unit of a microscope system (100) according to one of claims 1 to 14 a method according to one of claims 15 to 28 is carried out.

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