Microscopic imaging method

Abstract

The present disclosure relates to a method for preparing microscopic imaging, comprising: acquiring a plurality of images by a microscopic system and annotating each of the plurality of images with respective metadata relating to the microscopic system; and grouping the plurality of images, so that at least part of the images sharing the metadata in the plurality of images are grouped in the same image group.

Description

Technical Field

[0001] The present invention relates to a method for preparing microscopic images, a microscopic imaging method, a microscopic system, and the use of said system. Background Technology

[0002] Typically, when preparing for microscopic imaging, users capture microscope images with different hardware settings and compare the images to find the appropriate hardware settings for further use.

[0003] However, comparing the results of different settings is both time-consuming and error-prone, especially when settings need to be manually managed. For example, recovering previous settings from existing image data in order to capture more images with the same settings is very time-consuming. After preparation, the actual microscopic imaging is performed based on appropriate hardware settings selected according to the captured images.

[0004] Therefore, when preparing or planning microscopic imaging and / or during image processing, there is a need for more reliable, more ideal fail-safe and / or timely processing of microscope images. Summary of the Invention

[0005] The present invention solves this problem and relates to a method for preparing microscopic images. Claim 1 defines a method for preparing microscopic images, comprising (1) acquiring a plurality of images using a microscopic system and labeling each of the plurality of images with corresponding metadata related to the microscopic system, and (2) grouping the plurality of images such that images that at least partially share metadata are grouped into the same image group.

[0006] Metadata and images are interconnected, specifically in that images are grouped based on metadata, for example, with a one-to-one correspondence between groups and metadata. Grouping can refer to assigning images to groups (more specifically, assigning each image to a single group), such that images sharing the same metadata are grouped together. Therefore, a group can represent specific metadata and include multiple images. Grouping can also refer to placing images that at least share specific metadata into the same subcategory. Image groups can be presented visually or non-visually.

[0007] In this regard, images within the same image group share at least partially identical, and optionally completely identical, metadata. In other words, a group of images can be characterized by the fact that the contained images share at least partially identical, and optionally completely identical, metadata. Metadata can serve as the basis for defining different groups, each representing specific metadata. The metadata does not need to be completely different between different groups, and there can be overlap between the metadata characterizing different groups. Overlapping metadata between groups can be, for example, related to the microscope's hardware components (e.g., light source and / or detector) or other hardware settings, see below.

[0008] Multiple groups can be provided, for example, at least three groups, or at least five groups. The number of groups can vary, for example, depending on the complexity of the metadata.

[0009] Metadata can indicate microscope settings, such as microscope hardware settings, including specific settings like the intensity of excitation light applied when capturing an image. Therefore, when a group is characterized by a specific excitation light intensity, the excitation light intensity at the time of image acquisition can determine the group to which the image belongs.

[0010] Other parameters may include the wavelength of the light source, detection bands, lifetime settings, and the number of scans allocated to different detection bands during the acquisition of an image.

[0011] For fluorescence microscopy, specific exemplary hardware settings may include, for example, detection bands (e.g., including beam path (a list of integer values), start and / or end of image acquisition, ... markers and names), laser lines (e.g., including wavelength, beam path, index, intensity, relative intensity, name, abbreviation), and / or markers (dye, observed brightness, characteristic color, name, and optional other user-specific information, such as lab ID, date, comments, etc.). Accordingly, markers / dyes are, for example, part of the hardware settings.

[0012] If all the parameters of the hardware settings correspond to a specific (coloring) group, then the image belongs to that (coloring) group.

[0013] Images can be associated with the same sample, such that all groups are also associated with the same sample to be imaged by microscopy.

[0014] This invention provides various improved embodiments. In some embodiments, grouping images according to metadata can reduce errors compared to ungrouped images. In particular, metadata-based grouping helps to obtain images with the required metadata because grouping "guarantees" the exclusion of images that do not have specific metadata. Furthermore, it allows for rapid switching between different metadata (i.e., different groups) and corresponding images, as this can be achieved by switching between groups. By distinguishing between a first group and a second group, which define an "umbrella" for each image in each group, comparisons between images in the first group and images in the second group (i.e., images from different groups) can be simplified.

[0015] Alternatively or additionally, the present invention can simplify the comparison between different metadata and corresponding images (e.g., based on the hardware settings of the corresponding captured images). For example, since images with shared metadata are assigned to different groups, it can help to quickly compare images characterized by different metadata, which can help users “find” images with different metadata.

[0016] Alternatively or additionally, this can allow for improved processing of the captured images. For example, processing based on the same metadata (e.g., the same hardware setup) and corresponding visibility and visual presentation can be performed based on the provided set. This invention can support the creation and evaluation of microscope settings (e.g., hardware settings) for subsequent use based on metadata.

[0017] Otherwise, without the method of this invention, users might have to remember the settings used when acquiring images. Specifically, it might be impossible to identify (e.g., see) whether two images were captured using the same settings. If the settings were to be reused, they would need to be explicitly saved or re-imported from existing image data. Managing these settings would require user intervention and thus manual operation. Clearly, this is prone to errors when dealing with large amounts of sample data.

[0018] Optionally, the method includes adjusting the microscopy system (to a microscope setup configuration) based on metadata of the images in the group, and using said metadata (here, the microscope setup configuration) to optionally capture / acquire more images for the group. This facilitates capturing (i.e., acquiring) more images using the same hardware settings for the group. Specifically, after preparation, the actual microscopic imaging is performed based on appropriate metadata (e.g., hardware settings) characteristics of a group, which have been selected based on the captured images. In particular, a group can be selected based on the images assigned to that group to obtain, for example, optimal image quality.

[0019] Alternatively, the metadata can be loaded before adjusting the microscope system to the microscope setup configuration based on the metadata. This allows the microscope to be adapted to its configuration, especially automatically. The microscope setup configuration can involve the focus, light source brightness, and the sensitivity of sensors (scintillators, cameras, semiconductor sensors such as CCDs).

[0020] Optionally, the microscope settings configuration can be adjusted by the user to a user-defined microscope settings configuration. In particular, the user can change and further adjust the microscope settings configuration, for example, for an automatically suggested configuration.

[0021] Optionally, the microscope settings configuration and / or user-adjusted microscope settings configuration are adjustments based on microscope feedback, which may be provided by an optimization algorithm running on the microscope system, and more preferably used to optimize image quality, signal-to-noise ratio, and / or pixel saturation. Microscope feedback may relate to parameters such as light intensity, focus sharpness, sample contrast, temperature fluctuations, and / or stage drift. At least some or all of these parameters may affect image quality and can be used to automatically adjust settings for optimal imaging performance.

[0022] Optionally, the method also includes adding image groups to existing groups if images cannot be grouped into existing groups. For example, when the microscope's hardware settings are changed, another group is added to represent the group characterized by metadata reflecting the changed hardware settings.

[0023] Optionally, the method further includes: selecting a group (optionally, selecting images in the group) that needs to indicate the remaining images in the group. For example, selecting a group or selecting images in a group results in indicating the chronological order of the captured images.

[0024] Optionally, the method also includes selecting a group that indicates the images in that group. For example, and optionally, selecting a group could mean displaying the image history of captured images group by group.

[0025] Optionally, the method also includes excluding images from a particular group from the remaining groups. For example, images that do not belong to the corresponding group can be automatically excluded, for instance, by not visually displaying the excluded images. This helps reduce errors, especially when only images sharing common metadata (such as hardware settings) are available.

[0026] Optionally, the method also includes capturing multiple images, including imaging samples stained with different fluorophores. This represents a preferred embodiment and application of the method of the present invention. In other words, the method can be performed on multicolor / multiplex microscopy and / or fluorescence microscopy. In this respect, the images can be characterized by the fluorophores stained on the sample. A group can include images of samples stained with different fluorophores. Thus, a group can be characterized by images indicating different staining agents / dyes on the sample but indicating the same hardware setup. For example, 15 different fluorophores can be used. This can be referred to as a staining group.

[0027] One or more hardware settings can be grouped into a staining group. Each staining group has the same hardware settings.

[0028] Optionally, the method includes storing metadata in memory (cells), such as a storage medium. This allows for the rapid retrieval of different metadata when desired.

[0029] Optionally, the method includes reading metadata of the image and optionally storing the metadata in, for example, a memory. Reading the metadata of the image may optionally include reading metadata of previously acquired or imported images.

[0030] The present invention also relates to a microscopic imaging method, including the method for preparing microscopic images according to the present invention, which further includes processing images in groups. This may involve post-processing images based on groups. This facilitates including only images that share specific metadata for (post)processing. Thus, groups created during microscope preparation or planning can be associated with subsequent processing because the images are processed in groups.

[0031] In another alternative embodiment, images within a group can be processed based on metadata associated with the group. For example, hardware settings may be related to addressing spectral mixing issues during post-processing. Thus, shared hardware settings within a group can form the basis for processing images within that group.

[0032] In another alternative embodiment, image processing of the group may involve corrections based on metadata associated with the group, such as spectral correction. For example, when processing images from a fluorescence microscope, signal crosstalk can be reduced. Such correction may include demixing the image using a linear matrix based on hardware settings. One example is incorporating the fluorescence lifetime characteristics of the fluorophore. Another example is correction based on fluorescence lifetime and natural background (intrinsic fluorescence). This may involve using a time gate to select the detector signal based on photon arrival time, or separating the fluorescence component through FLIM (fluorescence lifetime imaging microscopy) attenuation fitting. Correction may also include denoising and deconvolution, which may be implemented in successive steps or combined with spectral or lifetime separation. Additionally or alternatively, a trained neural network may perform corrections using spectral and / or lifetime information from the acquired images. Metadata associated with the group may include parameters for these processing steps, such as a neural network model.

[0033] Optionally, the method includes acquiring images based on metadata and saving the metadata on which the image acquisition is based as image annotations and / or storing it in memory. This allows images to be acquired without confirmation or even knowledge of the microscope settings (i.e., microscope setting configuration) at the time of acquisition, because the metadata is stored in the image and / or memory, and thus makes the metadata available after image acquisition.

[0034] Optionally, the method includes retrieving metadata from memory and modifying the retrieved metadata, as well as creating training data based on the modified retrieved metadata to obtain improved metadata. This allows for training on the training data to obtain optimized metadata.

[0035] The present invention also relates to a microscopy system having a microscope and a computer, wherein the system is configured to perform the steps of the method of the present invention. Optionally, the computer includes a processor, which is further optionally configured to perform at least some of the steps of the present invention.

[0036] Optionally, the system is configured for use in fluorescence microscopy. In other words, the system can be configured for use in multicolor / multiplexing microscopy.

[0037] Optionally, the system or computer includes a memory for storing metadata.

[0038] Optionally, the system also includes a display device, such as a monitor or another graphical user interface, optionally for visual output, and more preferably for outputting grouping results.

[0039] Optionally, the system includes a user interface for initiating instructions on metadata, optionally via buttons included in the user interface. For example, pressing the button can enable a history function (e.g., to display all captured images) and / or to obtain metadata.

[0040] Optionally, the user interface can be configured to select options for adjusting metadata.

[0041] The present invention also relates to a fluorescence microscope, wherein the system is used to image samples stained with fluorophores under the microscope's hardware settings and to group samples stained with different fluorophores under the same hardware settings into the same group. This is particularly helpful in avoiding errors when processing images in multicolor / multiplexing microscopes.

[0042] The following describes detailed embodiments of the invention, as well as further related advantages and features, wherein these examples should not be considered as limiting the invention.

[0043] The term “and / or” as used in this document includes any and all combinations of one or more of the associated listed items and may be abbreviated to “ / ”.

[0044] Although some aspects are described in the context of an apparatus, they clearly also represent a description of the corresponding method, where a box or device corresponds to a method step or a feature of a method step. Similarly, aspects described in the context of method steps also represent a description of a corresponding box, item, or feature of the corresponding apparatus.

[0045] This method involves at least some steps performed by a computer, such as grouping multiple images. At least this step can be considered a computer-implemented step.

[0046] More generally, some or all of the method steps can be performed by (or using) hardware devices (such as processors, microprocessors, programmable computers, or electronic circuits). In some embodiments, one or more of the most important method steps can be performed by such devices.

[0047] Depending on specific implementation requirements, embodiments of the present invention can be implemented in hardware or software. This implementation can be achieved using non-transient storage media (e.g., digital storage media such as floppy disks, DVDs, Blu-ray discs, CDs, ROMs, PROMs, EPROMs, EEPROMs, or flash memory) that store electronically readable control signals that cooperate (or are capable of cooperating with) a programmable computer system to execute corresponding methods. Therefore, the digital storage media can be computer-readable. Attached Figure Description

[0048] Figure 1A microscopic system according to an embodiment of the present invention is illustrated schematically.

[0049] Figure 2 The structure of the microscopy system according to an embodiment of the present invention is illustrated schematically.

[0050] Figure 3(a) schematically illustrates the graphical user interface of the microscopy system according to an embodiment of the present invention.

[0051] Figure 3(b) schematically illustrates the graphical user interface of the microscopy system according to an embodiment of the present invention. Detailed Implementation

[0052] Figure 1 A microscopy system 100, including a computer (system) 102 and a microscope 101, is schematically shown. In this embodiment, the system 100 is configured for fluorescence microscopy and may also be a system for multicolor / multiplexing microscopy.

[0053] Figure 2 The structure 200, including metadata (here, microscope setup 201) and correspondingly grouped images 202, is schematically illustrated. The metadata 201 and image groups 202 are interconnected because the images are grouped based on the metadata 201. More generally, each metadata 202 is associated with a single image group 202.

[0054] Here, metadata 201 can be related to microscope settings. Specifically, metadata 201 can be related to multiple (here, four) different microscope (e.g., hardware) settings 203, 203a, 203b, and 203c, such as... Figure 2 As instructed. Furthermore, Figure 2 Four distinct image groups 202 are shown: a first group consisting of at least images 204, 204', and 204'' characterized by common metadata 203; a second group consisting of at least images 204a, 204a', and 204a'' characterized by common metadata 203a; a third group consisting of at least images 204b, 204b', and 204b'' characterized by common metadata 203b; and a fourth group consisting of at least images 204c, 204c', and 204c'' characterized by common metadata 203c. More or fewer images per group are possible.

[0055] Microscope settings 203, 203a, 203b, and 203c may share common parameters, such as those related to the light source and detector, but differ in specific parameters, such as the intensity used when acquiring images. Therefore, in this example, the first to fourth image groups 202 indicate different light intensities.

[0056] Accordingly, if the user selects image 204 from the first group defined by hardware setting 203, the user will receive instructions for the remaining images 204', 204'' in the first group. This can help the user quickly assess the suitability of hardware setting 203, for example, by taking all images 204, 204', and 204'' into account.

[0057] Alternatively, if the user selects the first group defined by hardware setting 203, the user will receive instructions for all images 204, 204', and 204'' in the first group. This can help the user quickly assess the suitability of hardware setting 203, for example, by taking all images 204, 204', and 204'' into account.

[0058] The grouping based on metadata 201 helps users process various images. For example, when a user analyzes a second group, defined by setting 203a and the corresponding images 204a, 204a', and 204a'', it helps the user make an evaluation because all images 204, 204', and 204'' in the first group are excluded.

[0059] Structure 200 can be created by acquiring images using the microscope system 100 and labeling each of the multiple images 204, ..., 204a, ..., 204b, ..., 204c, ... with corresponding metadata 203, 203a, 203b, 203c related to the microscope system 100. Subsequently, the multiple images are grouped such that images 204, 204', 204'' from the multiple images sharing the hardware setup 203 are grouped into the same image group. This operation continues until all images are grouped according to metadata 201. If an image cannot be grouped into an existing group, an image group can be added to the existing group.

[0060] Figures 3(a) and 3(b) schematically illustrate metadata and images associated with fluorescence microscopy, and more specifically, show stained samples displayed on the graphical user interface 300. Thus, Figures 3(a) and 3(b) illustrate a specific application of the invention for spectral mixing of fluorophores A, B, C, and D in fluorescence microscopy. In this case, multiple images are captured by imaging samples stained with different fluorophores A through D. Accordingly, samples can be stained with different fluorophores and imaged under the same or different hardware settings of microscope 101. Samples stained with different fluorophores under the same hardware settings are displayed in the graphical user interface 300.

[0061] To prepare the sample / sample, the staining protocol, antibodies, and other parameters were defined.

[0062] A set of hardware settings is created based on a list of markers (selected by the user) and an experimental execution speed (selected by the user). For example, a higher speed means fewer differential settings, and therefore higher crosstalk during image acquisition. Furthermore, staining groups representing groups associated with the dyes used are determined. These staining groups are generated by a computer using an algorithm to determine the appropriate (optimized) distribution of the desired settings.

[0063] Each staining group has one or more hardware settings. Acquired images are labeled with the hardware settings used for image acquisition (i.e., metadata). Images are assigned (i.e., grouped) to staining groups based on their hardware settings, which are the same for images corresponding to the same hardware settings.

[0064] In the example, metadata is loaded before adjusting the microscope system 100 to the microscope settings configuration based on metadata. Microscope settings configuration can relate to focal point, light source brightness, sensor (scintillator, camera, semiconductor sensor such as CCD, etc.) or the sensitivity of other entities. Users can adjust the microscope settings configuration, for example, by changing and further refining it.

[0065] Microscope settings configuration and / or user-adjusted microscope settings configuration are adjusted based on microscope feedback, which may optionally be provided by an optimization algorithm running on the microscope system 100, and more preferably used to optimize image quality, signal-to-noise ratio, and / or pixel saturation. Such feedback may be related to one or more parameters, such as light intensity, focus sharpness, sample contrast, temperature fluctuations, and / or stage drift.

[0066] More specifically, Figures 3(a) and 3(b) schematically illustrate the graphical user interface 300 used to present the groupings. All images 204, ..., 204'''' acquired using the same specific hardware settings 203 are summarized in the overview. This can be filtered according to various criteria.

[0067] Specifically, the light source and detector are the same in Figures 3(a) and 3(b). Other acquisition settings unrelated to processing may differ.

[0068] The hardware setup contains all the information needed to record a sample stained with a given set of fluorophores (e.g., 15). Each fluorophore is recorded by at least one detection band (channel) and excited with light of a defined wavelength. Each detection band will produce a pixel image; therefore, the “recorded image” is a set of pixel images. Since the microscope has a maximum of 5 detection units, it is impossible to record all 15 fluorophores in a single scan, so multiple scans are necessary. These hardware setups can be used to record samples stained with only a subset of fluorophores, such as one of the 15 fluorophores. The resulting “recorded image” still consists of 15 pixel images (channels), with a few (ideally one) including the signal. In Figure 3, each row corresponds to a sample stained only with dyes A, B, C… but all samples are still recorded with the same hardware setup used to process all 15 dyes (A, B, C, … “15”).

[0069] Metadata 201 is stored in memory (cell), such as storage medium 104. Metadata 201 of an image (optionally a previously acquired or imported image) is read and stored in memory 104.

[0070] Images can be acquired based on metadata 201, and the metadata used for image acquisition is saved / stored as image annotations and / or stored in memory 104. This allows for image acquisition without confirmation or even knowledge of the microscope settings (i.e., microscope setting configuration), because metadata 201 is stored in the image and / or memory, and thus makes the metadata available after image acquisition.

[0071] Metadata 201 can be retrieved from memory 104 and the retrieved metadata can be modified, therefore the system or computer includes memory for storing metadata.

[0072] System 100 or computer system 102 includes a memory 104 for storing metadata and a processor 103 for performing at least some steps of the method of the present invention.

[0073] System 100 includes a display device, such as a monitor or another graphical user interface 300, for visual output of groups, such as image group 202 and metadata 201.

[0074] The system includes a user interface 300 for initiating an instruction to metadata 201, optionally via a button 302 included in the user interface 300. For example, button 302 can be pressed to perform a history function (e.g., to display all acquired images) and / or to acquire metadata 201.

[0075] The user interface 300 is configured to select options for adjusting metadata 201 (such as hardware settings).

[0076] System 100 allows the creation of training data based on modified retrieved metadata to obtain improved metadata. This allows training on the training data to obtain optimized metadata.

[0077] The history list 301, as shown in Figure 3(b), allows switching between all hardware settings 203a, b, and c where image data is available, thus providing a method for quickly evaluating hardware settings 203a, b, and c. In the example shown in Figure 3(b), setting 203 has been selected, while settings 203a, b, and c are not. Accordingly, the graphical user interface 300 displays images 204, 204', 204'', ..., at different intensities for each fluorophore A through D, representing only the image group associated with setting 203, as indicated by the register for each fluorophore. These four intensities belong to one (i.e., the same) hardware setting. Changing one of the intensities implies a different hardware setting.

[0078] In the underlying data model, images are grouped according to metadata 201 (such as hardware settings used to capture images). Therefore, the user interface 300 displays the currently acquired images and helps simplify the processing / management of the captured images.

[0079] All images matching the current hardware settings (e.g., 203) in microscope 101 are displayed on interface 300 in an overview graphical format. All displayed images are labeled with the same hardware settings and correspond to the hardware settings used for the current image acquisition. If the microscope's hardware settings change, images labeled with the changed settings are checked, and if so, these images labeled with the changed hardware settings are displayed on the user interface. Images labeled with previous hardware settings are not displayed on the user interface (but are hidden).

[0080] By selecting an image, a specific image can be viewed and evaluated in the viewer. The history list 301 provides an overview of all existing hardware settings 203, 203a, 203b, 203c, and the image group 202 captured using these hardware settings. Entries may contain information such as the image acquisition time (e.g., date) and / or the capture time of the first and last images in the group. By selecting an entry in the history dialog box / list 301, the hardware settings 203 used can be set in the microscope 101, and the corresponding images 204, 204', 204'' will be displayed in the overview. This allows users to quickly switch between different settings and more easily capture new images using existing hardware settings. For example, the microscope 101 can be adjusted based on the metadata 201 of the images in the image group 202, and more images can be acquired for that image group using the metadata.

[0081] The number or number of staining groups displayed on the graphical interface 300 can depend on the unmixing strategy. For example, a staining group can be defined for each marker. Alternatively, similar markers can be grouped into one staining group. Further alternatively, only a single staining group including all markers can be defined.

[0082] This also makes it easier to use associated images in image group 202 to evaluate the corresponding metadata 201.

[0083] After image preparation / planning and acquisition, the images are processed in groups. More specifically, images in image group 203 are processed based on metadata 201 associated with the group. Since the group can reflect fluorescence imaging, image processing includes reducing fluorescence-related problems. More specifically, image processing within a group includes correction based on metadata 201 associated with that group 202, optionally spectral correction. Referring to Figures 3(a) and 3(b) and fluorescence microscopy, correction may include a linear matrix for unmixing images based on image group 202 (i.e., based on metadata 201) in conjunction with fluorophores. Here, relying on image group 202 is particularly helpful in avoiding errors in processing, such as including “erroneous” images in processing steps.

[0084] Another example is correction based on the fluorescence lifetime characteristics of a fluorophore, more specifically, correction based on fluorescence lifetime and natural background (intrinsic fluorescence). This could involve using time-gating to select detector signals based on photon arrival time, or separating the fluorescence components through FLIM (fluorescence lifetime imaging microscopy) attenuation fitting. Alternatively or additionally, correction could include denoising and deconvolution, which could be implemented in successive steps or combined with spectral or lifetime separation. Additionally or alternatively, a trained neural network could perform correction using spectral and / or lifetime information from the acquired images. Metadata associated with the group could include parameters for these processing steps, such as the neural network model.

[0085] Some embodiments relate to a microscope 101, which includes a reference Figure 1 In the described system. Alternatively, microscope 101 may be used as a reference. Figure 1 Part of the system 100 described, or connected to the system 100. Figure 1 A system 100 for performing the methods described herein is schematically illustrated. The system 100 includes a microscope 101 and a computer system 102. The microscope 101 is configured to capture images and is connected to the computer system 102. The computer system 102 is configured to perform at least a portion of the methods described herein. The computer system 102 and the microscope 101 may exist as separate entities or may be integrated into the same housing. The computer system 102 may be part of the central processing system of the microscope 101, and / or as part of a sub-component of the microscope 101 (such as a sensor, actuator, camera, or illumination unit, etc.).

[0086] Computer system 102 may be a local computer device (e.g., a personal computer, laptop, tablet, or mobile phone) having one or more processors and one or more storage devices; or it may be a distributed computer system (e.g., a cloud computing system having one or more processors and one or more storage devices distributed across various locations (e.g., at local clients and / or one or more remote server clusters and / or data centers)). Computer system 102 may include any circuitry or combination of circuitry. In one embodiment, computer system 102 may include one or more processors, which may be of any type. Here, "processor" may mean any type of computing circuitry, such as, but not limited to, a microprocessor for a microscope or microscope component (e.g., a camera), a microcontroller, a complex instruction set computing (CISC) microprocessor, a reduced instruction set computing (RISC) microprocessor, a very long instruction word (VLIW) microprocessor, a graphics processor, a digital signal processor (DSP), a multi-core processor, a field-programmable gate array (FPGA), or any other type of processor or processing circuitry. Other types of circuitry that may be included in computer system 102 may be custom circuitry, application-specific integrated circuits (ASICs), etc., such as one or more circuits (e.g., communication circuitry) used in wireless devices like mobile phones, tablets, laptops, two-way radios, and similar electronic systems. Computer system 102 may include one or more storage devices, which may include one or more storage elements suitable for a particular application, such as main memory in the form of random access memory (RAM), one or more hard disk drives, and / or one or more drives for processing removable media such as optical discs (CDs), flash memory cards, digital video discs (DVDs), etc. Computer system 102 may also include a display device, one or more speakers, and a keyboard and / or controller, which may include a mouse, trackball, touchscreen, voice recognition device, or any other device that allows a system user to input information into and receive information from computer system 102.

[0087] The present invention will be described in detail with reference to the embodiments shown in the accompanying drawings. Various variations and alternatives will be apparent to those skilled in the art based on the summary of the invention. All such variations and alternatives are part of the invention as long as they are covered by the appended claims.

[0088] List of reference numerals 100 Microscopic System 101 Microscope 102 Computer System 103 processor 104 Memory 200 Structure 201 metadata 202 Image Groups Hardware settings for models 203, 203a, 203b, and 203c Images 204 / ' / '', 204a / ' / '', 204b / ' / '', 204c / ' / '' 300 Graphical User Interface 301 History List Button 302.

Claims

1. A method for preparing a microscopic image, comprising: - Acquire multiple images using a microscope system (100), and annotate each of the multiple images with corresponding metadata (201) related to the microscope system (100), and - Group the plurality of images such that images that at least partially share metadata (201) are grouped into the same image group (202).

2. The method according to claim 1, further comprising: - Adjust the microscopy system (100) to the microscope settings configuration based on the metadata (201) of the images in the group (202), and use the microscope settings configuration to acquire further images for the optional group.

3. The method according to claim 2, wherein, Before adjusting the microscope system to the microscope settings configuration based on the metadata, load the metadata.

4. The method according to claim 2 or 3, wherein, The microscope settings are configured by the user to a user-defined microscope settings configuration.

5. The method according to claim 4, wherein, The microscope settings configuration is based on microscope feedback adjustment and may be provided by an optimization algorithm running on the microscope system, and more preferably for optimizing image quality, signal-to-noise ratio, and / or pixel saturation.

6. The method according to any one of the preceding claims further comprises: - If the image cannot be grouped into an existing group (202), add the image group to the existing group.

7. The method according to any one of the preceding claims further comprises: - When selecting an image in group (202), it is necessary to indicate the remaining images in the group.

8. The method according to any one of the preceding claims further comprises: - When selecting a group (202), you need to indicate the image in the group.

9. The method according to any one of the preceding claims further comprises: -Exclude the images in group (202) from the remaining groups.

10. The method according to any one of the preceding claims, wherein, - Acquire multiple images, including imaging samples stained with different fluorophores.

11. The method according to any one of the preceding claims, further comprising: - Store metadata in memory.

12. The method according to any one of the preceding claims further comprises: - Read the metadata of the image, optionally the metadata of previously acquired or imported images, and optionally store the metadata.

13. A microscopic imaging method, comprising the method of any one of the preceding claims, further comprising: - Process images by group.

14. The method of claim 13, further comprising: Images in a group are processed based on metadata associated with the group.

15. The method according to claims 13 and 14, further comprising: - The images in the processing group include correction based on metadata associated with the group, optionally spectral correction.

16. The method according to any one of claims 13 to 15, comprising: - Acquire images based on metadata, and save the metadata used for image acquisition as annotations for the images and / or store them in memory.

17. The method according to any one of claims 13 to 16, comprising: - Retrieve the metadata from the memory and modify the retrieved metadata, and create training data based on the modified retrieved metadata to obtain improved metadata.

18. A microscopic system (100) comprising a computer (102) and a microscope (101), the system optionally including a processor and configured to perform the steps of the method according to any one of the preceding claims.

19. The system according to claim 18, wherein, The system (100) is configured for use in fluorescence microscopy.

20. The system of claim 18 or 19 further includes a memory for storing metadata.

21. The system according to any one of claims 18 to 20 further includes a display device optionally used for visual output, and more preferably for grouped visual output.

22. The system according to any one of claims 18 to 21 further includes a user interface (300) for initiating instructions on metadata, optionally via buttons included in the user interface.

23. The system of claim 22, wherein the user interface (300) is configured to select options for adjusting metadata.

24. Use of the system (100) according to any one of claims 18 to 23 for fluorescence microscopy, wherein, The applications include imaging samples stained with fluorophores under microscope hardware settings, and grouping samples stained with different fluorophores under the same hardware settings into the same group.

Images