Optical imaging device for microscope

Abstract

To provide an optical imaging device for a microscope capable of precisely combining images generated in accordance with different imaging modes.SOLUTION: An optical imaging device comprises: a first optical system 108 for forming a first optical image corresponding to a first region of a sample 104 in accordance with a first imaging mode; a second optical system 112 for forming a second optical image corresponding to a second region of the sample in accordance with a second imaging mode; a memory for storing first distortion correction data, second distortion correction data, and transformation data; and a processor 124. The processor processes first image data representing the first optical image on the basis of the first distortion correction data, processes second image data representing the second optical image on the basis of the second distortion correction data, and combines the first and second distortion corrected image data on the basis of the transformation data for generating combined image data representing a combined image corresponding to a target region of an object.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to an optical imaging device for a microscope. Further, the present invention relates to a method for imaging a sample using a microscope, and a method for calibrating an optical imaging device for a microscope.

Background Art

[0002] In the field of microscopy, multiple different imaging modes are used to generate an optical image of a sample. Each of these imaging modes has advantages and disadvantages in terms of image quality, spatial resolution, imaging speed, exposure amount, etc. For example, in confocal imaging, higher spatial resolution is possible, but there is a drawback that the image acquisition time for a sample that requires point-by-point scanning for image generation becomes relatively long. In contrast, the wide-field imaging mode is advantageous in terms of the time required for image acquisition. However, the resolution of the image may be significantly reduced.

[0003] Therefore, when imaging the same sample, it is necessary to combine different modes. However, since different imaging modes may use different optical paths, it is a problem to match two images generated in different modes and to be able to display a properly combined image on a monitor, for example. When the user manually registers or aligns the images, a similar image structure is required to enable the registration. This is a cumbersome task, and accurate image fusion is often impossible.

[0004] In such a background, reference is made to European Patent No. 2322969, which discloses a microscope including a plurality of observation optical systems capable of acquiring images of the same sample in different modes. This microscope enables full utilization of the individual fields of view of the plurality of observation optical systems, thereby improving work efficiency. However, the aforementioned specification is not involved in automatically merging the images generated by the application of different imaging modes into a combined image.

Summary of the Invention

Problems to be Solved by the Invention

[0005] An object of the present invention is to provide an optical imaging device and method capable of accurately combining images generated according to different imaging modes. Further, this object is to provide a method for calibrating an optical imaging device for a microscope so that the optical imaging device can accurately combine images.

Means for Solving the Problems

[0006] The above problems are solved by the subject matter of the independent claims. Preferred embodiments are defined in the dependent claims and the following description.

[0007] According to one embodiment, an optical imaging device for a microscope includes a first optical system configured to form a first optical image corresponding to a first region of a sample according to a first imaging mode, and a second optical system configured to form a second optical image corresponding to a second region of the sample, wherein the first and second regions spatially coincide in a target region of the sample, and the first and second imaging modes are different from each other. The optical imaging device further includes a memory storing first distortion correction data suitable for correcting a first optical distortion caused by the first optical system in the first optical image, second distortion correction data suitable for correcting a second optical distortion caused by the second optical system in the second optical image, and conversion data suitable for correcting a positional shift between the first and second optical images. The optical imaging device further includes a processor configured to process first image data representing the first optical image based on the first distortion correction data to generate first distortion-corrected image data. This processor is further configured to process second image data representing the second optical image based on the second distortion correction data to generate second distortion-corrected image data. This processor is configured to combine the first and second distortion-corrected image data based on the conversion data to generate combined image data representing a combined image corresponding to the target region of the object.

[0008] The optical imaging device includes two optical systems, where each of these optical systems may have an image sensor adapted to a specific imaging mode. Each optical system is considered to potentially cause optical distortion in the optical image generated by this optical system. These first and second optical systems can use different optical paths for imaging such that the optical distortion induced by the first and second optical systems can be independent of each other. Thus, the first and second distortion correction data are not correlated with each other. The distortion correction data may be determined and stored independently for each optical system during assembly. Typically, the optical distortion induced by each optical system represents aberrations that cause blurring or distortion of the image such that proper alignment of the image is adversely affected. This becomes even more realistic since each optical distortion can be significantly different from each other for different imaging modes. Any such adverse effects can also be avoided by storing the first and second distortion correction data that are automatically considered when merging the first and second optical images into a combined image.

[0009] The memory of the optical imaging device further stores conversion data suitable for correcting the misalignment of positions between the first and second optical images. In contrast to the first and second distortion correction data that can be determined independently for each optical system, the conversion data represents data that takes into account the positional relationship between both optical systems, particularly the positional relationship between the optical systems that may cause misalignment of positions between the optical images.

[0010] The first and second optical systems are each used to image first and second regions of a sample, where these regions are spatially coincident in a target region of the sample. The first and second regions of the sample can be spatially coincident in different ways. For example, the first and second sample regions may be identical to each other, and thus the target region itself is likewise identical to each region. Alternatively, one of the sample regions may be completely contained within the other region, and thus the target region is formed by the contained sample region. Further alternatively, the first and second regions may partially overlap. In such a case, the target region is formed by an overlapping region common to both sample regions.

[0011] The optical imaging device may include a display unit on which a combined image corresponding to the imaged target region of the sample is displayed. Accordingly, the user can observe the target region of the sample based on the composite image that benefits from the advantages of both imaging modes.

[0012] Preferably, the first imaging mode is a wide-field mode and the second imaging mode is a confocal imaging mode. The composite image based on these fundamentally different imaging modes provides the user with image information far exceeding the reference image information.

[0013] In a preferred embodiment, the conversion data represents a positional shift between a first optical reference image formed by the first optical system according to the first imaging mode and a second optical reference image formed by the second optical system according to the second imaging mode. The optical reference images may be generated using a reference object adapted to be appropriately imaged in both imaging modes. Alternatively, live images generated during the actual imaging process may be used as the reference images.

[0014] Preferably, a calibration mode is provided in which the processor is configured to generate conversion data and store the conversion data in a memory before forming the first and second optical images. In this calibration mode, the processor may further be configured to generate first and second distortion correction data and store these data in the memory. Preferably, the calibration mode is applied to the manufacturing and assembly processes, so that the distortion correction data and the conversion data are already stored in the finished product. Therefore, the user does not need to worry about any calibration. Rather, the user can fully concentrate on experiments including sample preparation, adjustment of imaging parameters, etc.

[0015] The processor may be configured to determine correlation data representing the correlation between the first and second optical reference images and generate conversion data based on the correlation data. For example, an algorithm for determining a correlation coefficient based on the identification of structural features in the image may be applied. Based on this information, an iterative optimization procedure may be used to determine the required coordinate transformation.

[0016] The positional shift represented by the conversion data may include translation, rotation, scaling, shear, mirroring, and / or distortion.

[0017] In a preferred embodiment, the processor is configured to update the conversion data and store the updated conversion data in the memory. By updating the conversion data, the user can be made to be able to cope with changes that occur during the experiment. For example, changes due to drift, structural changes, dynamic processes of the sample, etc. can be corrected by re-determining the conversion stored in the memory.

[0018] As an example, the processor may be configured to cause a first optical system to generate a series of first optical images according to a first imaging mode, and a second optical system to generate a series of second optical images according to a second imaging mode. To re-determine the transformation, the processor may further be configured to determine a first tracking mark within one of the first optical images and a second tracking mark within one of the second optical images. In such a case, the processor is configured to perform tracking of the first tracking mark and the second tracking mark and update the transformation data based on the tracking. By automatically defining so-called landmarks in the form of the aforementioned tracking marks in the initial image and tracking these landmarks over time, it is possible to re-calibrate the transformation without reusing the calibration reference in the form of the reference object.

[0019] The processor may further be configured to combine the first and second distortion-corrected image data based on the transformation data such that one of the first and second optical images is mapped to a reference system defined by the other of the first and second optical images, or such that both optical images are mapped to a common reference system. In other words, a coordinate transformation is applied thereby to transform the coordinate system of one optical image to the coordinate system of the other image, or to transform the coordinate systems of both optical images to a new common coordinate system.

[0020] The first and second optical systems may be installed in a fixed positional relationship with respect to each other. Preferably, the two different imaging modes are adapted to be as closely as possible to each other by physical alignment of the respective optical elements. Such alignment allows minimizing the required transformation of the image points. Thus, interference caused by interpolation, rotation, etc. can be significantly reduced.

[0021] According to another aspect, a method for imaging a sample using a microscope is provided. This imaging method includes the following steps. That is, forming a first optical image corresponding to a first region of the sample according to a first imaging mode using a first optical system; forming a second optical image corresponding to a second region of the sample using a second optical system, wherein the first and second regions spatially coincide in the target region of the sample, and the first and second imaging modes are different from each other; obtaining first distortion correction data suitable for correcting a first optical distortion caused by the first optical system in the first optical image; obtaining second distortion correction data suitable for correcting a second optical distortion caused by the second optical system in the second optical image; obtaining conversion data suitable for correcting a positional shift between the first and second optical images; processing first image data representing the first optical image based on the first distortion correction data to generate first distortion-corrected image data; processing second image data representing the second optical image based on the second distortion correction data to generate second distortion-corrected image data; and combining the first and second distortion-corrected image data based on the conversion data to generate combined image data representing a combined image corresponding to the target region of the object.

[0022] According to another aspect, a method for calibrating an optical imaging device for a microscope is provided, the optical imaging device including a first optical system configured to form a first optical image corresponding to a first region of a sample according to a first imaging mode, a second optical system configured to form a second optical image corresponding to a second region of the sample, wherein the first and second regions spatially coincide in a target region of the sample and the first and second imaging modes are different from each other, a processor, and a memory. The method includes the following calibration steps. That is, obtaining first distortion correction data suitable for correcting a first optical distortion caused by the first optical system in the first optical image, obtaining second distortion correction data suitable for correcting a second optical distortion caused by the second optical system in the optical image, obtaining conversion data suitable for correcting a positional shift between the first and second optical images, and storing the first distortion correction data, the second distortion correction data, and the conversion data in the memory so as to be accessible by the processor.

[0023] This calibration method may be performed during the manufacturing and assembly process, so that the assembled product can be provided to the user with the required calibration data already stored therein.

[0024] According to a preferred embodiment, a first optical reference image of a reference object is formed using the first optical system according to the first imaging mode. A second optical reference image of the reference object is formed using the second optical system according to the second imaging mode. The conversion data is determined based on the positional shift between the first and second reference images. By using a single calibration reference in the above-described reference object form, the conversion data can be determined in a simple and reproducible manner. This calibration reference can also be used to determine the first and second distortion correction data.

[0025] By way of mere example, the reference object may include a grid formed by a plurality of spots. This grid is adapted to the first and second optical systems such that each optical system can image at least two of the plurality of spots over the magnification range available to it. Such a grid is used to ensure that the calibration reference contains sufficient structural information to achieve the required accuracy. In particular, the grid is formed to represent the same structure in both imaging modes. Further, the grid may be imaged in both transmitted light and fluorescence.

[0026] Hereinafter, specific embodiments will be described based on the drawings.

Brief Description of the Drawings

[0027]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Modes for Carrying Out the Invention

[0028] FIG. 1 shows a microscope 100 that includes an optical imaging device 102 configured to image a sample 104 on a microscope stage 106 according to different imaging modes. For example, this optical imaging device 102 can serve to image the sample 104 in a wide-field imaging mode and a confocal imaging mode. Needless to say, these imaging modes should be understood as examples only. Any other mode may be applied as long as these modes are adapted to provide image information that can be well combined for the generation of a composite image.

[0029] The optical imaging device 102 includes a first optical system 108 configured to form a first optical image corresponding to a first region 210 (see FIG. 2) of the sample 104 according to a first imaging mode, which is a wide-field imaging mode in this embodiment. Similarly, the optical imaging device 102 includes a second optical system 112 configured to form a second optical image corresponding to a second region 214 (see FIG. 2) of the sample 104 according to a second imaging mode, which is a confocal imaging mode in this embodiment. As schematically shown in FIG. 1, the first and second optical systems 108, 112 use different optical paths 116 and 118, respectively. Along these optical paths 116, 118, the detection light emitted from the sample 104 propagates through the optical systems 108, 112 and towards the image sensors 120, 122 respectively coupled to the optical systems 108, 112. The first image sensor 120 assigned to the first optical system 108 may be formed by a camera suitable for wide-field imaging. The second image sensor 122 assigned to the second optical system 112 may be formed by a sensor suitable for confocal imaging, such as a point detector.

[0030] The optical imaging device 102 further includes a processor 124 that can play a role in controlling the overall operation of the optical imaging device 102. In particular, this processor 124 is configured to process first image data representing a first optical image and second image data representing a second optical image, where the first and second optical images are respectively generated on image sensors 120, 122 using first and second optical systems 108, 112. For this purpose, the processor 124 is connected to the optical systems 108, 112 and the image sensors 120, 122 via control lines 126, 128, 130, 132.

[0031] The optical imaging device further includes a memory 134 connected to the processor 124 via a control line 136. Further, a display unit 138 connected to the processor 124 via a control line 140 may be provided.

[0032] Furthermore, a beam splitter or any other suitable light deflector 142 may be included in the optical imaging device 102 to provide different optical paths 116, 118 towards the first and second optical systems 108, 112.

[0033] As described above, the optical imaging device 102 can operate in a wide-field imaging mode and a confocal imaging mode to image the first region 210 and the second region 214 of the sample 104. The first and second regions 210, 214 spatially coincide in the target region of the sample 104 indicated by the hatched region 242 in FIG. 2. This spatial coincidence may be realized in different ways. For example, the second region 214 assigned to the confocal imaging mode may be completely contained within the first region, as shown in FIG. 2(a). Since this target region 242 is formed by the overlap of the first and second regions 210, 214, this target region 242 is the same as the second region 214 in the example shown in FIG. 2(a). Further, FIG. 2(b) shows an example in which the first and second regions 210, 214 are the same, and thus the target region 242 is the same as each region. In the example shown in FIG. 2(c), the first and second regions 210, 214 are partially overlapped, and thus the target region 242 is formed by the overlapping region common to both regions 210, 214. Needless to say, the spatial coincidence between the first region and the second regions 210, 214 is not limited to the examples shown in FIG. 2.

[0034] In the embodiment shown in FIG. 1, the memory 134 is provided for storing first distortion correction data and second distortion correction data. The first distortion correction data is suitable for correcting first optical distortion, such as optical aberration, caused by the first optical system 108 when generating a first optical image of the first region 210 of the sample 104 according to the wide-field imaging mode. Similarly, the second distortion correction data serves to correct second optical distortion caused by the second optical system 112 when generating a second optical image of the second region 214 of the sample 104 according to the confocal imaging mode. Since the first and second optical systems 108, 112 use different optical paths 116, 118 for imaging the respective regions 210, 214, the optical distortions induced by the first and second optical systems 108, 112 are independent of each other. Therefore, the first and second distortion correction data can be determined independently and stored in the memory 134.

[0035] Furthermore, the memory 134 stores conversion data that can be used to correct the positional deviation between the first and second optical images respectively created in the wide-field imaging mode and the confocal imaging mode. The first and second distortion correction data may be assigned independently to each optical system 108, 112, but the conversion data stored in the memory 134 reflects the positional relationship between the optical systems 108, 112.

[0036] Processor 124 utilizes the first and second distortion correction data, as well as the conversion data stored in memory 134, for providing a combined image corresponding to the target region 242 of target 104. This combined image provides image information obtained from both wide-field imaging and confocal imaging. To create the combined image, the processor processes the first image data representing the first optical image based on the first distortion correction data to generate the first distortion-corrected image data. Similarly, processor 124 processes the second image data representing the second optical image based on the second distortion correction data to generate the second distortion-corrected image data. Then, based on the conversion data, the processor combines the first and second distortion-corrected image data to generate combined image data representing the combined image to be displayed on display unit 138.

[0037] Optical imaging device 102 may provide a calibration mode in which processor 124 generates conversion data and stores these data in memory 134. This calibration mode is preferably applied to the manufacturing and assembly process of optical imaging device 102, and thus can be automatically used at a later stage when the user operates microscope 100 for imaging sample 104. The first and second distortion correction data, which are independently assigned to each optical system 108, 112, may also be generated by processor 124 in the calibration mode.

[0038] A reference object may be used to calibrate optical imaging system 102. By way of mere example, such a reference object may be formed by grid 350 as shown in FIG. 3.

[0039] The grid 350 includes a plurality of spots 352 provided in a rectangular array. This grid 350 is adapted to the first and second optical systems 108, 112, so that both optical systems 108, 112 can image at least two of the plurality of spots 352 over the available magnification range of the microscope 100 despite the fact that the optical systems 108, 112 apply different imaging modes.

[0040] For calibration purposes, the first optical system 108 generates a first optical reference image of the grid 350 in a wide-field imaging mode. Accordingly, the second optical system 112 generates a second optical reference image of the grid 350 in a confocal imaging mode. Subsequently, the processor 124 generates first and second distortion correction data and stores these data in the memory 134. Further, the processor determines the positional shift between the first and second reference images representing the grid 350. Based on this shift, the processor 124 generates transformation data and stores these data in the memory 134.

[0041] The use of a reference object as shown in FIG. 3 for calibration of the optical imaging device 102 should be understood as merely illustrative. Thus, the transformation data may be generated based on live images created when the user operates the microscope 100 for imaging of the sample 104. Further, the live images may be used to update the initial transformation data stored in the memory 134 for correction of drifts, structural changes, dynamic processes of the sample, etc. occurring during the experiment. For example, the processor 124 can cause each of the optical systems 108, 112 to generate a series of optical images according to each imaging mode. For each sequence, the processor determines tracking marks 452 as shown in FIG. 4 within this series of initial images and tracks the tracking marks 452 over time, i.e., over a plurality of images generated subsequent to the initial image. By using the tracking marks 452, the processor 124 can generate updated transformation data for recalibrating the transformation without the need to use a reference object as shown in FIG. 3.

[0042] To generate the conversion data, the processor 124 may be configured to determine correlation data representing the correlation between the reference images. As described above, an image representing the reference object or a live image may be used.

[0043] The flowchart of FIG. 5 shows a method for calibrating the optical imaging system 102 according to one embodiment.

[0044] In step S2, the first optical system 108 images the reference object as shown in FIG. 3 to form a first reference image. The image data representing the first reference image may be stored in an image memory (not shown in the figure). In step S4, the processor 124 determines the first distortion correction data by comparing, for example, the digital data representing the first reference image with nominal data predefined to correspond to an ideal reference image, i.e., an image generated without any optical distortion. In step S6, the processor 124 stores the first distortion correction data in the memory 134.

[0045] In step S8, the second optical system 112 forms a second reference image, and the processor 124 stores the corresponding image data in the image memory. In step S10, the processor 124 determines the second distortion correction data. In step S12, the processor 124 stores this second distortion correction data in the memory 134. Steps S8, S10, and S12 for generating and storing the second distortion correction data are performed in the same manner as steps S2, S4, and S6 related to the first distortion correction data.

[0046] In step S14, the processor 124 determines the conversion data based on the first and second reference images as described above. Finally, in step S16, the processor 124 stores these conversion data in the memory 134.

[0047] The flowchart shown in FIG. 6 shows a method for imaging the sample 104 according to one embodiment.

[0048] In step S12, the first optical system 108 forms a first optical image of the first target region 210 according to the wide-field imaging mode. Accordingly, in step 14, the second optical system 112 forms a second optical image of the second region 214 according to the confocal imaging mode. As described above with reference to FIG. 2, the first and second regions of the sample 104 spatially coincide in the target region 242. The first and second image data representing the first and second optical images are stored in the image memory in steps 12 and 14.

[0049] After the first and second images are generated, the processor 124 reads the first distortion correction data from the memory 134 in step S16. Similarly, in step S18, the processor 124 reads the second distortion correction data from the memory 134. In step S20, the processor 124 reads the conversion data from the memory 134.

[0050] In step S22, the processor 124 processes the first image data representing the first optical image based on the first distortion correction data to create the first distortion-corrected image data. Accordingly, in step 24, the processor 124 processes the second image data representing the second optical image based on the second distortion correction data to generate the second distortion-corrected image data. The first and second distortion-corrected image data are stored in the image memory.

[0051] Finally, in step S26, the processor 124 combines the first and second distortion-corrected image data using the conversion data read from the memory 134. Thus, combined image data representing the combined image is generated. The combined image corresponding to the target region 242 of the sample 104 may be displayed on the display unit 138. Further, the combined data may be stored in an image memory not shown in the figure.

[0052] Figure 7 schematically shows a specific transformation executed by processor 124 to combine the first and second optical images. According to the example of Figure 7, processor 124 uses two mapping operations T’ and T’’ to transform a first coordinate system KS1 assigned to the first image and a second coordinate system KS2 assigned to the second image into a third coordinate system KS3. The third coordinate system KS3 represents a common reference system assigned to the combined image formed from the first and second images.

[0053] Needless to say, the transformation shown in Figure 7 is merely exemplary. Thus, processor 124 can combine the first and second distortion-corrected image data based on the transformation data such that the first coordinate system KS1 is mapped to the second coordinate system KS2, or such that the second coordinate system KS2 is mapped to the first coordinate system KS1.

[0054] Although some aspects have been described in the context of an apparatus, it is clear that these aspects also represent a description of corresponding methods, where a block or device corresponds to a step or a feature of a step. Similarly, aspects described in the context of a step also represent a description of corresponding blocks or items or features of a corresponding apparatus. Some or all of the steps may be performed by, for example, a hardware device (or using a hardware device) such as a processor, a microprocessor, a programmable computer, or an electronic circuit. In some embodiments, any one or more of the extremely important steps may be performed by such a device.

[0055] Depending on certain implementation requirements, embodiments of the present invention can be implemented in hardware or software. This implementation is executable by a non-transitory recording medium, which is a digital recording medium, such as a floppy disk, DVD, Blu-ray, CD, ROM, PROM, EPROM, EEPROM, or FLASH memory, that stores electronically readable control signals that cooperate (or are capable of cooperating) with a programmable computer system to implement each method. Thus, the digital recording medium may be computer-readable.

[0056] Some embodiments of the present invention include a data carrier having electronically readable control signals that can cooperate with a programmable computer system so that any of the methods described herein are implemented.

[0057] Generally, embodiments of the present invention can be implemented as a computer program product comprising program code, which operates to implement any of the methods when the computer program product is executed on a computer. This program code may be stored, for example, on a machine-readable carrier.

[0058] Another embodiment includes a computer program stored on a machine-readable carrier for implementing any of the methods described herein.

[0059] Thus, in other words, embodiments of the present invention are computer programs having program code for implementing any of the methods described herein when the computer program is executed on a computer.

[0060] Accordingly, another embodiment of the present invention is a recording medium (or data carrier or computer-readable medium) containing a computer program stored therein for implementing any of the methods described herein when executed by a processor. The data carrier, digital recording medium or recording medium is typically tangible and / or non-transitory. Another embodiment of the present invention is an apparatus as described herein comprising a processor and a recording medium.

[0061] Accordingly, another embodiment of the present invention is a data stream or signal sequence representing a computer program for implementing any of the methods described herein. The data stream or signal sequence may be configured to be transferred, for example, via a data communication connection such as the Internet.

[0062] Another embodiment includes a processing means, for example, a computer or programmable logic device configured or adapted to implement any of the methods described herein.

[0063] Another embodiment includes a computer having an installed computer program for implementing any of the methods described herein.

[0064] Another embodiment of the present invention includes an apparatus or system configured to transfer (e.g., electronically or optically) a computer program for implementing any of the methods described herein to a receiver. The receiver may be, for example, a computer, a mobile device, a storage device, etc. The apparatus or system may include, for example, a file server for transferring the computer program to the receiver.

[0065] In some embodiments, a programmable logic device (e.g., a field programmable gate array) may be used to execute some or all of the functions of the methods described herein. In some embodiments, a field programmable gate array may cooperate with a microprocessor to implement any of the methods described herein. Generally, and advantageously, the methods may be implemented by any hardware device.

Explanation of Signs

[0066] 100 Microscope 102 Optical imaging device 104 Sample 106 Microscope stage 108 First optical system 112 Second optical system 116 Optical path 118 Optical path 120 Image sensor 122 Image sensor 124 Processor 126 Control line 128 Control line 130 Control line 132 Control line 134 Memory 136 Control line 138 Display unit 140 Control line 142 Optical deflector 210 First region 214 Second region 242 Target region 350 Grid 352 Spot 452 Tracking mark KS1 First coordinate system KS2 Second coordinate system KS3 Third coordinate system T Transformation operation T’ Transformation operation

Claims

1. An optical imaging device (102) for a microscope (100), wherein the optical imaging device (102) comprises: A first optical system (108) configured to form a first optical image corresponding to a first region (210) of a sample (104) according to a first imaging mode; A second optical system (112) configured to form a second optical image corresponding to a second region (214) of the sample (104) according to a second imaging mode, wherein the first region (210) and the second region (214) spatially coincide in a target region (242) of the sample (104), and the first and second imaging modes are different from each other; A memory (134) storing first distortion correction data suitable for correcting a first optical distortion caused by the first optical system (108) in the first optical image, second distortion correction data suitable for correcting a second optical distortion caused by the second optical system (112) in the second optical image, and conversion data suitable for correcting a positional shift between the first and second optical images; A processor (124); Including, The processor (124) is configured to: Process first image data representing the first optical image based on the first distortion correction data to generate first distortion-corrected image data; Process second image data representing the second optical image based on the second distortion correction data to generate second distortion-corrected image data; Combine the first and second distortion-corrected image data based on the conversion data to generate combined image data representing a combined image corresponding to the target region (242) of the sample (104); The first region (210) and the second region (214) are identical; Optical imaging device (102).

2. The first imaging mode is a wide-field imaging mode; The second imaging mode is a confocal imaging mode; The optical imaging device (102) according to Claim 1.

3. The conversion data represents a positional shift between a first optical reference image formed by the first optical system (108) according to the first imaging mode and a second optical reference image formed by the second optical system (112) according to the second imaging mode. The optical imaging device (102) according to claim 1 or 2.

4. A calibration mode is provided in which the processor (124) is configured to generate the conversion data and store the conversion data in the memory (134) before forming the first and second optical images. The optical imaging device (102) according to any one of claims 1 to 3.

5. The processor (124) is configured to determine correlation data representing the correlation between the first and second optical reference images and generate the conversion data based on the correlation data. The optical imaging device (102) according to claim 3 or 4.

6. The positional shift represented by the conversion data includes translation, rotation, scaling, shearing, mirroring, and / or distortion. The optical imaging device (102) according to any one of claims 1 to 5.

7. The processor (124) is configured to update the conversion data and store the updated conversion data in the memory (134). The optical imaging device (102) according to any one of claims 1 to 6.

8. The processor (124) is configured to cause the first optical system (108) to generate a sequence of first optical images according to the first imaging mode, and cause the second optical system (112) to generate a sequence of second optical images according to the second imaging mode. The processor (124) is configured to determine a first tracking mark (452) within one of the first optical images and determine a second tracking mark within one of the second optical images. The processor (124) is configured to perform tracking of the first tracking mark and the second tracking mark (452) and update the conversion data based on the tracking. The optical imaging device (102) according to claim 7.

9. The processor (124) is configured to combine the first and second distortion-corrected image data based on the conversion data such that one of the first and second optical images is mapped to a reference system (KS1, KS2) defined by the other of the first and second optical images, or such that both optical images are mapped to a common reference system (KS3). The optical imaging device (102) according to any one of claims 1 to 8.

10. The first and second optical systems (108, 112) are installed in a fixed positional relationship with respect to each other. The optical imaging device (102) according to any one of claims 1 to 9.

11. A method for imaging a sample (104) using a microscope (100), the method comprising: Forming a first optical image corresponding to a first region (210) of the sample (104) according to a first imaging mode using a first optical system (108); Forming a second optical image corresponding to a second region (214) of the sample (104) according to a second imaging mode using a second optical system (112), wherein the first region (210) and the second region (214) spatially coincide in a target region (242) of the sample (104), and the first and second imaging modes are different from each other; Obtaining first distortion correction data suitable for correcting a first optical distortion caused by the first optical system (108) in the first optical image; Obtaining second distortion correction data suitable for correcting a second optical distortion caused by the second optical system (112) in the second optical image; Obtaining conversion data suitable for correcting a positional shift between the first and second optical images; Processing first image data representing the first optical image based on the first distortion correction data to generate first distortion-corrected image data; Processing second image data representing the second optical image based on the second distortion correction data to generate second distortion-corrected image data; Combining the first and second distortion-corrected image data based on the conversion data to generate combined image data representing a combined image corresponding to the target region (242) of the sample (104); comprising The first region (210) and the second region (214) are identical. Method.

12. A method for calibrating an optical imaging device (102) for a microscope (100), the optical imaging device (102) comprising: A first optical system (108) configured to form a first optical image corresponding to a first region (210) of a sample (104) according to a first imaging mode; A second optical system (112) configured to form a second optical image corresponding to a second region (214) of the sample (104) according to a second imaging mode, wherein the first region (210) and the second region (214) spatially coincide in a target region (242) of the sample (104), and the first and second imaging modes are different from each other, the second optical system (112); A processor (124); A memory (134); Including; The method includes the following calibration steps, namely: Obtaining first distortion correction data suitable for correcting a first optical distortion caused by the first optical system (108) in the first optical image; Obtaining second distortion correction data suitable for correcting a second optical distortion caused by the second optical system (112) in the second optical image; Obtaining conversion data suitable for correcting a positional shift between the first and second optical images; Storing the first distortion correction data, the second distortion correction data, and the conversion data in the memory (134) so as to be accessible by the processor (124); Including; The first region (210) and the second region (214) are the same. Method.

13. A first optical reference image of a reference object (350) is formed using the first optical system (108) according to the first imaging mode, A second optical reference image of the reference object is formed using the second optical system (112) according to the second imaging mode, The conversion data is determined based on a positional shift between the first and second reference images. The method according to claim 12.

14. The reference object includes a grid (350) formed by a plurality of spots (352), and the grid (350) is adapted to the first and second optical systems (108, 112) such that at least two of the plurality of spots (352) can be imaged over a range of magnifications available to each optical system (108, 112). The method according to claim 13.

15. A computer program comprising program code for performing the method according to any one of claims 11 to 14 when the computer program is executed on a processor.

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