Beam image shift data acquisition method based on camera continuous exposure recording

Abstract

This application relates to the field of cryo-electron microscopy and discloses a method for acquiring beam image migration data based on continuous exposure recording by a camera. The method includes: determining the total exposure time for continuously capturing multiple target points in a single exposure; controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time; during the continuous capture of multiple target points in a single exposure, applying beam-image migration technology to switch the target points, and initiating electron beam blocking during the switching process; acquiring a multi-point, multi-frame file containing image information of multiple target points and electron beam blocking information during the switching process; and extracting imaging data of each target point from the multi-point, multi-frame file based on the position of the electron beam blocking information. This application improves data acquisition efficiency.

Description

Technical Field

[0001] This disclosure relates to the field of cryo-electron microscopy, and more specifically, to a method for acquiring beam image migration data based on continuous exposure recordings from a camera. Background Technology

[0002] In the field of cryo-electron microscopy, direct electron probe cameras (such as the K3 or Falcon4i) have become the mainstream data acquisition equipment for single particle analysis (SPA) and cryo-electron tomography (Cryo-ET).

[0003] Currently, cryo-electron microscopy (cryo-EM) utilizes beam image migration to acquire single-particle data and electron tomography sequence tilt data. This method significantly reduces the time required to move the sample stage during data acquisition, greatly improving data acquisition efficiency. However, direct electron detectors (DEMs) require data acquisition for each target point separately, meaning each target point needs exposure. After each exposure, sensor readout time (SRT), data buffering, and hardware synchronization operations are required, resulting in an actual recording time (ART) that is significantly longer than the set exposure time (SET), thus severely reducing data acquisition efficiency. In summary, in beam image migration data acquisition, an ART greater than the set exposure time becomes a key issue limiting data acquisition efficiency. Summary of the Invention

[0004] In view of the above situation, this application provides a method for acquiring beam image offset data based on continuous exposure recording of a camera, which aims to solve the above problems or at least partially solve the above problems.

[0005] In a first aspect, embodiments of this application provide a method for acquiring beam image migration data based on continuous exposure recordings from a camera, the method comprising:

[0006] The total exposure time for capturing multiple target points in a single continuous exposure is determined; the camera is controlled to capture multiple target points in a single continuous exposure based on the total exposure time; during the process of the camera capturing multiple target points in a single continuous exposure, beam-image shift technology is applied to switch the target points, and beam occlusion is initiated during the switching process; a multi-point multi-frame file containing image information of multiple target points and beam occlusion information during the switching process is acquired, and imaging data of each target point is extracted from the multi-point multi-frame file based on the position of the beam occlusion information.

[0007] In some embodiments, determining the total exposure time for capturing multiple target points in a single continuous exposure includes: determining the total exposure time for capturing multiple target points in a single continuous exposure based on the exposure time of a single target point, the beam obstruction time when switching target points, and the number of target points.

[0008] In some embodiments, before controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time, the method further includes: determining the slope of the linear fitting curve corresponding to the camera capturing the target points at different frame rates, wherein the linear fitting curve is a linear fitting curve of the set exposure time and the actual exposure time; selecting the frame rate range corresponding to the target slope within a preset slope range; and controlling the target camera's target frame rate to be within the frame rate range.

[0009] In some embodiments, controlling the target frame rate of the target camera to be within the frame rate range includes: determining the base exposure time of a single target point based on the total dose and the single-point dose of a single target point; and controlling the target frame rate of the target camera to be within the frame rate range based on the base exposure time and camera parameters.

[0010] In some embodiments, extracting imaging data for each target point from the multi-point, multi-frame file based on the position of the beam occlusion information includes:

[0011] Determine the illumination intensity of each frame in the multi-point, multi-frame file; identify frames below the light intensity threshold as beam occlusion frames; using beam occlusion frames as separators, combine consecutive frames above the light intensity threshold into one image file, and output the target images of multiple target points respectively.

[0012] In some embodiments, for cryo-electron computed tomography (cryo-electron computed tomography) data, each tilt angle corresponds to a multi-point, multi-frame file; the method further includes generating a data file containing an image name, tilt angle, and exposure parameters for each multi-point, multi-frame file.

[0013] In some embodiments, when acquiring data via cryo-electron tomography, the method further includes: controlling the tilt of the sample stage during a camera delay time, such that the sample stage movement and the camera delay time are executed in parallel.

[0014] Secondly, embodiments of this application also provide a beam image migration data acquisition device based on continuous exposure recording of a camera. The device includes: a calculation module for determining the total exposure time for continuously capturing multiple target points in a single exposure; a control module for controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time; during the process of continuously capturing multiple target points in a single exposure, applying beam-image migration technology to switch the target points, and initiating beam occlusion during the switching process; an acquisition module for acquiring a multi-point multi-frame file containing image information of multiple target points and beam occlusion information during the switching process; and a processing module for extracting imaging data of each target point from the multi-point multi-frame file based on the position of the beam occlusion information.

[0015] In some embodiments, the calculation module is specifically used to determine the total exposure time for a single exposure to continuously capture multiple target points based on the exposure time of a single target point, the beam occlusion time when switching target points, and the number of target points.

[0016] In some embodiments, before controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time, the calculation module is further configured to determine the slope of the linear fitting curve corresponding to the camera capturing the target points at different frame rates, wherein the linear fitting curve is a linear fitting curve of the set exposure time and the actual exposure time; select the frame rate range corresponding to the target slope within the preset slope range; and control the target frame rate of the target camera to be within the frame rate range.

[0017] In some embodiments, the calculation module is specifically used to determine the base exposure time of a single target point based on the total dose and the single-point dose of a single target point; and to control the target frame rate of the target camera to be within the frame rate range based on the base exposure time and camera parameters.

[0018] In some embodiments, the processing module is configured to determine the illumination intensity of each frame in a multi-frame file; identify frames below a light intensity threshold as beam occlusion frames; and combine consecutive frames above the light intensity threshold into an image file, using beam occlusion frames as separators, and output target images of multiple target points respectively.

[0019] In some embodiments, for cryo-electron computed tomography (cryo-electron computed tomography) data, each tilt angle corresponds to a multi-point, multi-frame file; the processing module is further configured to generate a data file containing image name, tilt angle, and exposure condition parameters for each multi-point, multi-frame file.

[0020] In some embodiments, when acquiring data via cryo-electron computed tomography, the control module is further configured to control the tilt of the sample stage during the camera delay time, so that the sample stage movement and the camera delay time are executed in parallel.

[0021] Thirdly, embodiments of this application also provide an electronic device, including: a processor; and a memory arranged to store computer-executable instructions, which, when executed, cause the processor to perform the steps described in the first aspect.

[0022] Fourthly, embodiments of this application also provide a computer-readable storage medium that stores one or more programs, which, when executed by an electronic device including multiple applications, cause the electronic device to perform the steps described in the first aspect.

[0023] The at least one technical solution adopted in this application embodiment can achieve the following beneficial effects: by determining the total exposure time for continuously capturing multiple target points in a single exposure, the camera is controlled to continuously capture multiple target points in a single exposure based on the total exposure time. During the capture of multiple target points, when switching the target point being captured using beam-image shift technology, the camera is controlled to initiate beam occlusion, acquiring multi-frame files containing image information of multiple target points and beam occlusion information when switching target points. Through the beam-image shift data acquisition method recorded by continuous camera exposure, multiple acquisitions at multiple points are integrated into a single continuous exposure, requiring only one camera delay time cost, avoiding the camera delay time present during each single-point capture and significantly improving data acquisition efficiency. Attached Figure Description

[0024] The accompanying drawings, which are included to provide a further understanding of this application and form part of this application, illustrate exemplary embodiments and are used to explain this application, but do not constitute an undue limitation of this application. In the drawings:

[0025] Figure 1 A flowchart illustrating the beam image offset data acquisition method for continuous exposure recording of a camera provided in this application embodiment is shown.

[0026] Figure 2 This illustration shows a schematic diagram of a multi-frame file output from a single continuous exposure provided in an embodiment of this application;

[0027] Figure 3 This illustration shows a comparison diagram of total exposure time provided in an embodiment of this application;

[0028] Figure 4 A flowchart illustrating the single-point exposure time calibration method provided in an embodiment of this application is shown;

[0029] Figure 5 A schematic diagram of the linear fitting curve provided in an embodiment of this application is shown;

[0030] Figure 6 This illustration shows a schematic diagram of the multi-frame file processing procedure provided in an embodiment of this application;

[0031] Figure 7 This invention provides a structural diagram of a beam image offset data acquisition device for continuous exposure recording of a camera, according to an embodiment of this application.

[0032] Figure 8 A schematic diagram of the structure of an electronic device provided in an embodiment of this application is shown. Detailed Implementation

[0033] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions of this application will be clearly and completely described below in conjunction with specific embodiments and corresponding drawings. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of them. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0034] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this application are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such use can be interchanged where appropriate so that the embodiments of this application described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the term "comprising" and its variations should be interpreted as open-ended terms meaning "including but not limited to."

[0035] Cryo-ET data acquisition requires collecting low-dose images of the same target region at different consecutive tilt angles. Repeated stage tilting and stabilization consumes a significant portion of the data acquisition time. In recent years, to improve Cryo-ET data acquisition throughput, some studies have employed beam image shift (BIS) technology, which allows for the simultaneous acquisition of multiple target points at the same tilt angle during a single continuous tilt, thus improving data acquisition efficiency by saving the time spent on repeated stage tilting. Nevertheless, the acquisition speed of traditional Cryo-ET combined with BIS (approximately 300 tomographic images per day) is still far lower than that of Single Particle Analysis (SPA) methods (approximately 300-400 micrographs per hour).

[0036] When using direct electron detection (DDD) cameras such as the K3 or Falcon4i for data acquisition, the actual recording time (ART) is longer than the set exposure time (SET) due to the camera delay time (CDT) after each exposure. The camera delay is primarily caused by sensor readout time (SRT), which allows the camera's sensor to transfer image data from the sensor chip to a computer for storage or processing after exposure. SRT depends on the camera type, resolution, and frame rate. For example, a K3 camera might require several milliseconds to read a frame. However, actual testing shows that the linear fit between SET and CDT has an intercept term (approximately 2 seconds). This intercept term represents other factors affecting CDT, such as data buffer time and latency between control software and hardware. The CDT between each exposure significantly reduces data acquisition efficiency.

[0037] Based on this, the present invention proposes a beam image migration data acquisition method for continuous camera exposure recording. By combining single uninterrupted continuous exposure with beam-image migration technology, multi-point acquisition is integrated into a single exposure. Only one camera delay time cost needs to be paid, avoiding the camera delay time that exists when the camera is exposed after each single-point shooting, and greatly improving the data acquisition efficiency.

[0038] The present application will now be described in detail through specific embodiments.

[0039] Figure 1 This paper illustrates a flowchart of a method for acquiring beam image shift data from continuous exposure recordings using a camera, as provided in an embodiment of this application. Figure 1 It can be seen that this application includes at least steps S101-S105:

[0040] Step S101: Determine the total exposure time for a single exposure to continuously capture multiple target points.

[0041] Step S102: Control the camera to continuously capture multiple target points in a single exposure based on the total exposure time.

[0042] The camera can be a direct electronic detection camera such as K3 or Falcon4i, or other cameras with the same camera exposure method as described in the background of this application. The embodiments of this application are not specifically limited here.

[0043] Step S103: During the process of continuously shooting multiple target points in a single exposure of the camera, the beam-image shift technology is applied to switch the shooting target points, and beam occlusion is activated during the switching process.

[0044] In this context, beam obstruction can be understood as a black frame, meaning that the camera cannot capture images of the switching process while the target point is being switched.

[0045] Step S104: Obtain a multi-point, multi-frame file containing image information of multiple target points and beam occlusion information during the switching process.

[0046] like Figure 2 As shown, the obtained file includes image frames of three target points, as well as beam occlusion frames when switching target points.

[0047] Step S105: Based on the position of the beam occlusion information, extract the imaging data of each target point from the multi-point multi-frame file.

[0048] from Figure 1 As can be seen from the method shown, this application combines single continuous exposure with beam-image shift technology to switch between target points, integrating multi-point acquisition into a single exposure, requiring only one camera delay time cost, such as... Figure 3 As shown, in the prior art, each single-point data acquisition is followed by a CDT after SET. However, the beam image offset data acquisition method based on continuous camera exposure recording provided in this application integrates the exposure times of multiple single points together, and the total exposure time of multiple single points corresponds to a CDT. This avoids the camera delay time that exists when the camera is exposed after each single-point shooting, and greatly improves the data acquisition efficiency.

[0049] In some embodiments of this application, in the above method, the total exposure time in step S101 is determined based on the exposure time of a single target point, the beam occlusion time when switching target points, and the number of target points. Specifically, the total exposure time is calculated based on the following formula: Total exposure time = (exposure time of a single target point + beam occlusion time) * number of target points.

[0050] Experiments have shown that the proposed beam image migration data acquisition method based on continuous camera exposure recording achieves the most significant improvement in acquisition efficiency when the exposure time for a single target point is within a preset range. Therefore, it is necessary to calibrate the single-point exposure time of the camera before calculating the total exposure time.

[0051] Specifically, such as Figure 4 As shown, the calibration process for single-point exposure time is as follows:

[0052] Step S201: Determine the slope of the linear fitting curve corresponding to the camera capturing the target point at different frame rates.

[0053] The linear fitting curve is a linear fitting curve between the set exposure time and the actual exposure time. Specifically, multiple exposures are performed with different SETs to obtain the corresponding ARTs for different SETs. The linear fitting curve between the SET and the ART is plotted as f(x) = ax + b, where f(x) is the ART, x is the SET, a represents the slope, and b is the intercept term representing the factors affecting CDT.

[0054] Different frame rates will affect the slope of the linear fit, such as Figure 5 As shown, the linear fitting curves are presented for frame rates of 0.02-0.12 s / frame. At low frame rates, the slope of the linear fitting curve is close to 1. In this case, only the intercept term affects the ART, so the longer the total exposure time, the more significant the efficiency improvement should be. At higher frame rates, since the slope term is not 1, it means that extending the total exposure time increases the CDT beyond just the intercept term. For example, if the slope term is 1, and the SET is 100 s, the ART is only 102 s; but if the slope term is 2, and the SET is 100 s, the ART becomes 202 s. In this case, the improvement of the continuous exposure method is limited.

[0055] Step S202: Select the frame rate range corresponding to the target slope within the preset slope range.

[0056] Step S203: Control the target camera's target frame rate to be within the frame rate range.

[0057] Specifically, the base exposure time for a single target point is determined in advance based on the total dose and the single-point dose of a single target point, and the target frame rate of the target camera is controlled within the frame rate range based on the base exposure time.

[0058] For example, if the total electron dose of a single target point is determined to be 30e-, the target frame number is 30 frames, and the target frame rate is 0.1s / frame, then by adjusting the camera's Dose rate, C2 capture, spot number, and other parameters, the single-point exposure time can be made to 3s.

[0059] In this embodiment of the application, by adjusting the frame rate of the camera, the data acquisition efficiency is significantly improved.

[0060] In some embodiments of this application, step S105 may be specifically implemented as steps S301-S303, such as... Figure 6 As shown, it includes the following steps:

[0061] Step S301: Determine the illumination intensity of each frame image in the multi-point multi-frame file.

[0062] Step S302: Frames with light intensity below the light intensity threshold are identified as beam occlusion frames.

[0063] Step S303: Using the beam occlusion frame as a separator, combine consecutive frames with light intensity above the threshold into one image file, and output the target images of multiple target points respectively.

[0064] In some embodiments, for single-particle analysis type data processing, a Python script is used to split and output the multi-point dataset. The main operations include: setting a light intensity threshold, statistically analyzing the light intensity of each frame, marking frames below the threshold as beam occlusion frames, and using beam occlusion frames as separators to combine consecutive frames above the threshold into a single image file for output; that is, splitting the multi-point data file into single-point data files for output. The script can utilize CPU parallel processing or GPU acceleration and can be combined with MotionCorr2 to output a motion-corrected .mrc file.

[0065] In other embodiments, for cryo-electron computed tomography (cryo-electron computed tomography) data, the cryo-electron computed tomography data is acquired by tilting the sample stage in the target area, collecting a dataset containing a series of different tilt angles. At each tilt angle, a beam image migration data acquisition method based on continuous camera exposure recording can be applied to collect data. Further, a Python script is used to split and output the multi-point dataset. The main operations include: setting a light intensity threshold, statistically analyzing the light intensity of each frame, marking frames below the set threshold as beam occlusion frames, and using beam occlusion frames as separators, combining consecutive frames above the threshold into a single communication file for output, i.e., splitting the multi-point data file into single-point data files for output. In one embodiment, a Python script generates an mdoc file corresponding to the tilt dataset, containing basic data acquisition parameters, image names, and corresponding tilt angles, which can be recognized by subsequent data processing software.

[0066] In addition, to address the long delay caused by the high frame rate slope, during electron tomography data acquisition, it takes time to rotate the sample stage to different angles and then to stabilize after reaching the desired position. Since the software controlling the camera (e.g., SerialEM) is stuck, this embodiment uses an additional Python script to control the tilting of the sample stage during CDT. After CDT, the next set of images can be taken directly, thus saving some of the CDT caused by the high frame rate slope.

[0067] In some embodiments of this application, a continuous data acquisition device is provided, which corresponds one-to-one with the beam image shift data acquisition method based on continuous camera exposure recording in the above embodiments. For example... Figure 7 As shown, the beam image offset data acquisition device based on continuous exposure recording of a camera includes a calculation module 101, a control module 102, an acquisition module 103, and a processing module 104.

[0068] Calculation module 101 is used to determine the total exposure time for a single exposure to continuously capture multiple target points;

[0069] Control module 102 is used to control the camera to continuously capture multiple target points in a single exposure based on the total exposure time; during the process of the camera continuously capturing multiple target points in a single exposure, beam-image shift (BIS) technology is applied to switch the target points to be captured, and beam occlusion is activated during the switching process;

[0070] The acquisition module 103 is used to acquire multi-frame files containing image information of multiple target points and beam occlusion information during the switching process;

[0071] The processing module 104 extracts the imaging data of each target point from the multi-point multi-frame file based on the position of the beam occlusion information.

[0072] In some embodiments of this application, in the above-described apparatus, the calculation module 101 is specifically used to determine the total exposure time for a single exposure to continuously capture multiple target points based on the exposure time of a single target point, the beam obstruction time when switching target points, and the number of target points.

[0073] In some embodiments of this application, in the above-described apparatus, before controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time, the calculation module 101 is further configured to determine the slope of the linear fitting curve corresponding to the camera capturing the target points at different frame rates, wherein the linear fitting curve is a linear fitting curve of the set exposure time and the actual exposure time; select the frame rate range corresponding to the target slope within the preset slope range; and control the target frame rate of the target camera to be within the frame rate range.

[0074] In some embodiments of this application, in the above-described apparatus, the calculation module 101 is specifically used to determine the base exposure time of a single target point based on the total dose and the single-point dose of a single target point; and to control the target frame rate of the target camera to be within the frame rate range based on the base exposure time and camera parameters.

[0075] In some embodiments of this application, in the above-described apparatus, the processing module 104 is specifically used to determine the illumination intensity of each frame image in a multi-point multi-frame file; determine frames below the light intensity threshold as beam occlusion frames; and combine consecutive frames above the light intensity threshold into an image file, using the beam occlusion frames as separators, and output target images of multiple target points respectively.

[0076] In some embodiments of this application, in the above-described apparatus, for cryo-electron tomography data, each tilt angle corresponds to a multi-point multi-frame file; the processing module 104 is also used to generate a data file containing image name, tilt angle and exposure condition parameters for each multi-point multi-frame file.

[0077] In some embodiments of this application, when acquiring data via cryo-electron computed tomography, the control module 102 is also used to control the tilt of the sample stage during the camera delay time, so that the sample stage movement and the camera delay time are executed in parallel.

[0078] It should be noted that any of the aforementioned beam image migration data acquisition devices based on continuous camera exposure records can implement the aforementioned beam image migration data acquisition method based on continuous camera exposure records one by one, which will not be elaborated here.

[0079] Figure 8 A schematic diagram of the structure of an electronic device provided in an embodiment of this application is shown. Figure 8 As shown, at the hardware level, this electronic device includes a processor, and optionally also includes an internal bus, a network interface, and memory. The memory may include main memory, such as high-speed random-access memory (RAM), or it may include non-volatile memory, such as at least one disk drive. Of course, this electronic device may also include other hardware required for other business operations.

[0080] The processor, network interface, and memory can be interconnected via an internal bus, which can be an ISA (Industry Standard Architecture) bus, a PCI (Peripheral Component Interconnect) bus, or an EISA (Extended Industry Standard Architecture) bus, etc. The bus can be divided into address bus, data bus, control bus, etc. For ease of representation, Figure 8 The symbol is represented by a single double-headed arrow, but this does not mean that there is only one bus or one type of bus.

[0081] Memory is used to store programs. Specifically, programs may include program code, which includes computer operation instructions. Memory may include main memory and non-volatile memory, and provides instructions and data to the processor.

[0082] The processor reads the corresponding computer program from non-volatile memory into memory and then runs it, forming a beam image migration data acquisition device based on continuous exposure recording of a camera at the logical level. The processor executes the program stored in memory and specifically performs the aforementioned method.

[0083] The processor may be an integrated circuit chip with signal processing capabilities. In implementation, each step of the above method can be completed by integrated logic circuits in the processor's hardware or by instructions in software form. The processor can be a general-purpose processor, including a Central Processing Unit (CPU), a Network Processor (NP), etc.; it can also be a Digital Signal Processor (DSP), an Application Specific Integrated Circuit (ASIC), a Field-Programmable Gate Array (FPGA), or other programmable logic devices, discrete gate or transistor logic devices, or discrete hardware components. It can implement or execute the methods, steps, and logic block diagrams disclosed in the embodiments of this application. The general-purpose processor can be a microprocessor or any conventional processor. The steps of the methods disclosed in the embodiments of this application can be directly embodied in the execution of a hardware decoding processor, or executed by a combination of hardware and software modules in the decoding processor. The software modules can reside in random access memory, flash memory, read-only memory, programmable read-only memory, electrically erasable programmable memory, registers, or other mature storage media in the art. The storage medium is located in the memory, and the processor reads the information in the memory and, in conjunction with its hardware, completes the steps of the above method.

[0084] This electronic device can execute the beam image migration data acquisition method based on continuous camera exposure recording provided in several embodiments of this application, and is implemented as a beam image migration data acquisition device based on continuous camera exposure recording. Figure 7 The functions of the embodiments shown are not described in detail here.

[0085] This application also proposes a computer-readable storage medium that stores one or more programs, the programs including instructions that, when executed by an electronic device including multiple applications, enable the electronic device to perform the beam image migration data acquisition method based on continuous camera exposure recording provided in several embodiments of this application.

[0086] Those skilled in the art will understand that embodiments of this application can be provided as methods, systems, or computer program products. Therefore, this application can take the form of a completely hardware embodiment, a completely software embodiment, or an embodiment combining software and hardware aspects. Furthermore, this application can take the form of a computer program product embodied on one or more computer-usable storage media (including but not limited to disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.

[0087] This application is described with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of this application. It will be understood that each block of the flowchart illustrations and / or block diagrams, and combinations of blocks in the flowchart illustrations and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, special-purpose computer, embedded processor, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, generate instructions for implementing the flowchart... Figure 1 One or more processes and / or boxes Figure 1 A device that provides the functions specified in one or more boxes.

[0088] These computer program instructions may also be stored in a computer-readable storage medium that can direct a computer or other programmable data processing device to function in a particular manner, such that the instructions stored in the computer-readable storage medium produce an article of manufacture including instruction means, which are implemented in a process Figure 1 One or more processes and / or boxes Figure 1 The function specified in one or more boxes.

[0089] These computer program instructions may also be loaded onto a computer or other programmable data processing equipment to cause a series of operational steps to be performed on the computer or other programmable equipment to produce a computer-implemented process, thereby providing instructions that execute on the computer or other programmable equipment for implementing the process. Figure 1 One or more processes and / or boxes Figure 1 The steps of the function specified in one or more boxes.

[0090] In a typical configuration, a computing device includes one or more processors (CPU), input / output interfaces, network interfaces, and memory.

[0091] Memory may include non-persistent storage in computer-readable media, such as random access memory (RAM) and / or non-volatile memory, such as read-only memory (ROM) or flash RAM. Memory is an example of computer-readable media.

[0092] Computer-readable media includes both permanent and non-permanent, removable and non-removable media that can store information using any method or technology. Information can be computer-readable instructions, data structures, modules of programs, or other data. Examples of computer storage media include, but are not limited to, phase-change memory (PRAM), static random access memory (SRAM), dynamic random access memory (DRAM), other types of random access memory (RAM), read-only memory (ROM), electrically erasable programmable read-only memory (EEPROM), flash memory or other memory technologies, CD-ROM, digital versatile optical disc (DVD) or other optical storage, magnetic tape, magnetic disk storage or other magnetic storage devices, or any other non-transferable medium that can be used to store information accessible by a computing device. As defined herein, computer-readable media does not include transient computer-readable media, such as modulated data signals and carrier waves.

[0093] It should also be noted that the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0094] Those skilled in the art will understand that embodiments of this application can be provided as methods, systems, or computer program products. Therefore, this application can take the form of a completely hardware embodiment, a completely software embodiment, or an embodiment combining software and hardware aspects. Furthermore, this application can take the form of a computer program product embodied on one or more computer-usable storage media (including, but not limited to, disk storage, CD-ROM, optical storage, etc.) containing computer-usable program code.

[0095] The above are merely embodiments of this application and are not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. A method of beam image shift data acquisition based on camera continuous exposure recording, characterized in that, The method includes: Determine the total exposure time for a single exposure to continuously capture multiple target points; Control the camera to continuously capture multiple target points in a single exposure based on the total exposure time; During the process of continuously shooting multiple target points in a single exposure of the camera, the beam-image shift technology is used to switch the shooting target points, and beam occlusion is activated during the switching process; Acquire a multi-point, multi-frame file containing image information of multiple target points and beam occlusion information during the switching process; Based on the location of the beam occlusion information, the imaging data of each target point is extracted from a single multi-point multi-frame file; Based on the position of the beam occlusion information, the imaging data of each target point is extracted from the multi-point multi-frame file, including: Determine the illumination intensity of each frame in a multi-point, multi-frame file; Frames with light intensity below the light intensity threshold are identified as beam occlusion frames; Using frames with light beam occlusion as separators, consecutive frames with light intensity exceeding the light intensity threshold are combined into a single image file, and target images of multiple target points are output separately.

2. The method according to claim 1, characterized in that, Determining the total exposure time for continuously capturing multiple target points in a single exposure includes: Based on the exposure time of a single target point, the beam occlusion time when switching target points, and the number of target points, the total exposure time for continuously shooting multiple target points in a single exposure is determined.

3. The method according to claim 1, characterized in that, Before controlling the camera to continuously capture multiple target points in a single exposure based on the total exposure time, the method further includes: Determine the slope of the linear fitting curve corresponding to the camera capturing the target point at different frame rates. The linear fitting curve is a linear fitting curve of the set exposure time and the actual exposure time. Select the frame rate range corresponding to the target slope within the preset slope range; Control the target camera's target frame rate to be within the frame rate range.

4. The method according to claim 3, characterized in that, The control of the target camera's target frame rate being within the frame rate range includes: The base exposure time for a single target point is determined based on the total dose and the single-point dose of the single target point. Based on the aforementioned basic exposure time and camera parameters, the target frame rate of the target camera is controlled to be within the frame rate range.

5. The method according to claim 1, characterized in that, For cryo-electron computed tomography (cryo-electron computed tomography) data, each tilt angle corresponds to a multi-point, multi-frame file; The method further includes: For each multi-point, multi-frame file, generate a data file containing the image name, tilt angle, and exposure condition parameters.

6. The method according to claim 5, characterized in that, When acquiring data via cryo-electron computed tomography, the method further includes: The sample stage is tilted during the camera delay time, so that the sample stage movement and the camera delay time are executed in parallel.

7. A beam image migration data acquisition device based on continuous camera exposure recording, characterized in that, The device includes: The calculation module is used to determine the total exposure time for capturing multiple target points in a single exposure. The control module is used to control the camera to continuously capture multiple target points in a single exposure based on the total exposure time; during the process of the camera continuously capturing multiple target points in a single exposure, the beam-image shift technology is applied to switch the target points to be captured, and beam blocking is activated during the switching process; The acquisition module is used to acquire multi-point, multi-frame files containing image information of multiple target points and beam occlusion information during the switching process; The processing module extracts imaging data for each target point from the multi-point, multi-frame file based on the position of the beam occlusion information. The processing module is specifically used to determine the illumination intensity of each frame in the multi-point, multi-frame file; to determine frames below the light intensity threshold as beam occlusion frames; and to combine consecutive frames above the light intensity threshold into one image file, using beam occlusion frames as separators, and output target images of multiple target points respectively.

8. An electronic device, comprising: processor; as well as A memory configured to store computer-executable instructions, characterized in that, when executed, the executable instructions cause the processor to perform the steps of the beam image migration data acquisition method based on continuous camera exposure recording as described in any one of claims 1-6.

9. A computer-readable storage medium storing one or more programs, characterized in that, When the one or more programs are executed by an electronic device including multiple applications, the electronic device performs the steps of the beam image migration data acquisition method based on continuous camera exposure recording as described in any one of claims 1-6.

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