Visual field leveling method for electric cylinder driven super-large visual field microscope

Abstract

The invention discloses a visual field leveling method for an electric cylinder driven super-large visual field microscope, which comprises an electric cylinder, a lens and a microscope objective table, the microscope objective table comprises an X axis and a Y axis and is used for controlling the deflection of the microscope objective table on the X axis and the Y axis, and the microscope objective table is provided with a shot sample. Comprising the following steps: S1, carrying out microscopic image acquisition on a shot sample at an initial angle through a lens to obtain an initial microscopic image; s2, performing microscopic image acquisition of a plurality of angles from the starting angle from the X axis in a set deflection angle range to obtain a plurality of deflection angle microscopic images; s3, after each time of angle deflection, performing microscopic image registration and cutting on the deflection angle microscopic image according to the initial microscopic image; s4, calculating and obtaining the definition of the microscopic image of the initial angle and the plurality of angles; and S5, by taking the optimal angle of the X axis as a reference, repeating the steps S2 to S4 to adjust the optimal angle of the Y axis, and completing global leveling of the X axis and the Y axis.

Description

Technical Field

[0001] The invention relates to the field of microscopic image technology, and in particular to a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder. Background Art

[0002] In modern scientific research and technological applications, wide field microscopes are widely welcomed due to their unique advantages. This type of microscope is mainly used for macroscopic to microscopic structural analysis in the fields of materials science, life science, and medical research. Compared with traditional microscopes, wide field microscopes can provide a wider field of view without sacrificing resolution, allowing observers to obtain information about larger samples at one time, which is of great significance for understanding complex biological tissue structures or material surface characteristics. In addition, it also has the characteristics of high throughput and can quickly scan large sample areas, greatly improving experimental efficiency and data acquisition speed, which is particularly important for applications that require large-scale sample testing.

[0003] When using a wide field microscope for observation, field leveling is a crucial step. Due to the limited working distance of the microscope, the depth of focus decreases accordingly when the magnification increases, which means that only a very thin layer of the sample is in focus. Therefore, in order to ensure that all areas of interest in the entire field of view can be clearly imaged, the sample must be accurately leveled. Good leveling can not only improve image quality and reduce edge blur, but also avoid the loss of important information due to local defocus, thereby ensuring the accuracy and reliability of the data. Especially when performing quantitative analysis, such as particle size measurement or cell counting, the accuracy of field leveling is directly related to the validity of the final result.

[0004] The traditional manual leveling method, although it can do the job in some simple cases, has many limitations.

[0005] First, manual operation is time-consuming, especially when dealing with complex or large samples, which require repeated adjustments until satisfaction is achieved; second, human visual judgment is easily affected by subjective factors, and different operators may obtain different leveling results, resulting in poor repeatability; third, operators are prone to fatigue under long-term and high-intensity work, which in turn affects the leveling accuracy.

[0006] Therefore, improvements are needed, and a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder is proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder, so as to overcome the deficiencies in the prior art.

[0008] To achieve the above object, the present invention provides the following technical solutions: The present application discloses a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder, comprising an electric cylinder, a lens and a microscope stage, wherein the microscope stage comprises an X-axis and a Y-axis for controlling the deflection of the stage on the X-axis and the Y-axis, and a sample to be photographed is arranged on the microscope stage, comprising the following steps: S1: Capture a microscopic image of the sample at the starting angle through the lens to obtain a starting microscopic image; S2: starting from the starting angle, collecting microscopic images at several angles on the X-axis within a set deflection angle range, and obtaining several deflection microscopic images; S3: after each angle deflection, the deflected microscopic image is registered and cropped according to the starting microscopic image to ensure the accuracy of subsequent images; S4: Calculate the clarity of the microscopic images at the starting angle and several angles; S5: Based on the optimal angle of the X axis, repeat S2 to S4 to adjust the optimal angle of the Y axis to complete the global leveling of the XY axes.

[0009] Preferably, S2 includes collecting data from any one of the X-axis and the Y-axis, and returning to S2 in S5 is performed from the other one of the X-axis and the Y-axis.

[0010] Preferably, S1 includes the following contents: acquiring a region of interest of the sample being photographed, and taking the current position as a starting angle, collecting a microscopic image at the current angle.

[0011] Preferably, S2 comprises the following sub-steps: S21: acquiring a plurality of acquisition angles within a preset deflection angle range according to the starting angle; S22: collecting microscopic images at several collection angles; S23: Acquire an image sequence based on the microscopic images at several angles.

[0012] Preferably, S3 includes the following contents: since the photographed samples in the field of view move during the angle deflection process, each image needs to be translated and registered based on the starting angle as a reference, and the registered images are cropped to retain the common area.

[0013] Preferably, the translation registration in S3 includes the following contents: registering the microscopic image by using the matchTemplate function in OpenCV.

[0014] Preferably, S4 includes the following contents: for the cropped starting microscopic image and the offset microscopic image in S3, the total variation is calculated respectively by the height, width and row and column indexes of the images; under the premise of having the same foreground target, the larger the total variation, the higher the clarity.

[0015] Preferably, S5 includes the following contents: according to the calculation result of S4, extracting the deflected microscopic image corresponding to the clearest image, and deflecting the microscope stage to the angle; then returning to S2, repeating the contents of S2 to S4 on the Y axis, obtaining the clearest angles on the X axis and the Y axis, and then realizing global field of view leveling in the XY two-dimensional dimensions.

[0016] The present application also discloses a field of view leveling device for an ultra-large field of view microscope driven by an electric cylinder, comprising a memory and one or more processors, wherein the memory stores executable code, and when the one or more processors execute the executable code, they are used to implement the above-mentioned field of view leveling method for an ultra-large field of view microscope driven by an electric cylinder.

[0017] The present application also discloses a computer-readable storage medium on which a program is stored. When the program is executed by a processor, the above-mentioned field of view leveling method for an ultra-large field of view microscope driven by an electric cylinder is implemented.

[0018] Beneficial effects of the present invention: (1) The field of view of a wide-field microscope is very large, and manual focusing is difficult to ensure that an ideal focal plane is obtained in the entire field of view. The present invention uses an automated electric cylinder leveling system to accurately level the entire large field of view, ensuring the consistency of imaging quality from the center to the edge. (2) In biological research, it is often necessary to image and analyze large-area tissue sections or cell samples, which needs to be done under a large field of view. The use of automated leveling in the present invention can greatly improve the efficiency of this process, achieve true high-throughput, unattended processing, and speed up the analysis process of samples with a large field of view. (3) For large field of view imaging, manual leveling is a very arduous task, which can easily cause visual fatigue to the operator, thereby increasing the risk of misoperation. The use of automated leveling in the present invention can reduce this burden and improve the reliability of experimental data.

[0019] The features and advantages of the present invention will be described in detail through embodiments in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 It is a flow chart of the steps of a method for leveling the field of view of a super-large field of view microscope driven by an electric cylinder according to the present invention; Figure 2 It is a schematic diagram of a single-dimensional leveling process of the present invention; Figure 3 It is a schematic diagram of the registration and cutting before and after of the present invention; Figure 4 It is a schematic diagram of the device of the present invention. DETAILED DESCRIPTION

[0021] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below through the accompanying drawings and embodiments. However, it should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention.

[0022] By integrating advanced sensor technology or computer vision algorithms, the present invention can realize a fast and accurate leveling process through an automated system. It not only greatly shortens the preparation time and improves work efficiency, but also significantly reduces human errors and ensures that each leveling reaches the best state. Therefore, automated leveling is not only an inevitable trend of technological progress, but also the key to improving scientific research quality and production efficiency.

[0023] See also Figure 1 The embodiment of the present invention provides a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder, which uses multiple real-time images of the sample as a data source and uses the optimal search concept to find the global clearest image to achieve the purpose of leveling.

[0024] The embodiments of the present invention include the following parts: The cylinder-driven microscope stage can be simplified into deflection around the X-axis and deflection around the Y-axis. The following steps take the X-axis as an example. The single-dimensional leveling process diagram is as follows Figure 2 shown.

[0025] S1. Microscopic image acquisition at the starting point: Find the ROI (region of interest) of the sample being photographed, and use the angle of the electric cylinder drive platform in the x direction at this time as the starting angle , collect the microscopic image at this angle ; S2. Multi-angle microscopic image acquisition: Send instructions to the electric cylinder driver through the program to let it is the starting angle, in the setting angle range In preset angle steps Perform multi-angle microscopic image acquisition to obtain image sequences ,in , This is because the starting angle has been collected in S1 The microscopic image does not need to be collected again at this step. Set the deflection angle range to 1.5° is (-1°, 1°), angle step The configuration of 0.2° is used as an example case. The image sequence obtained at this time is n=(2*1 / 0.2+1)-1=10; S3. Multi-angle microscopic image registration: Since the sample in the field of view will move slightly during the angle deflection process, the image sequence needs to be Each image in the image is a microscopic image with the starting point As a benchmark, image translation and registration are performed to ensure that subsequent calculations are based on and The registration is performed in the public field of view. The matchTemplate function in OpenCV can be used for registration. All registered images are cropped and only the public area is retained for subsequent calculations. The schematic diagram of the effect before and after registration and cropping is shown in the figure. Figure 3 As shown; In a feasible embodiment, after completing the registration and cropping, the image is processed with a uniform resolution, so as to reduce the time and resources required for processing in the subsequent processing and calculation process.

[0026] S4. Global clarity calculation: For the cropped and , respectively calculated and TV (total variation), under the premise of having the same foreground target, the larger the TV value, the more drastic the pixel change between adjacent pixels. In terms of visual perception, the more obvious the distinction between tissues is, the clearer it is. Therefore, the TV value can be used as the basis for its clarity. The TV calculation process can be expressed as formula (1): (1) Among them, h and w represent the pixel height and width of the cropped image, respectively, and i and j represent the row and column indexes of the image, respectively; S5. Calculation result feedback: After obtaining the TV calculation result of S4, the corresponding deflection angle can be obtained according to the position index with the largest TV value. The deflection angle value is fed back to the electric cylinder driver to drive the stage to deflect to the specified angle. , and take the microscopic image corresponding to this angle as the starting image , then perform the above steps S2 to S4 on the Y axis, and finally Feedback is given to the electric cylinder driver to deflect it to the specified angle to complete the global field of view leveling of the sample in the XY dimensions.

[0027] An embodiment of the field of view leveling device for an ultra-large field of view microscope driven by an electric cylinder of the present invention can be applied to any device with data processing capability, and the device with data processing capability can be a device or apparatus such as a computer. The device embodiment can be implemented by software, or by hardware or a combination of software and hardware. Taking software implementation as an example, as a device in a logical sense, the processor of any device with data processing capability reads the corresponding computer program instructions in the non-volatile memory into the internal memory and runs them. From the hardware level, if Figure 4 As shown, it is a hardware structure diagram of a field leveling device for a super-large field microscope driven by an electric cylinder according to the present invention, in which any device with data processing capability is located, except Figure 4 In addition to the processor, memory, network interface, and non-volatile memory shown, any device with data processing capabilities in the embodiment of the device may also include other hardware according to the actual function of the device with data processing capabilities, which will not be described in detail. The implementation process of the functions and effects of each unit in the above device is specifically detailed in the implementation process of the corresponding steps in the above method, which will not be described in detail here.

[0028] For the device embodiment, since it basically corresponds to the method embodiment, the relevant parts can refer to the partial description of the method embodiment. The device embodiment described above is only schematic, wherein the units described as separate components may or may not be physically separated, and the components displayed as units may or may not be physical units, that is, they may be located in one place, or they may be distributed on multiple network units. Some or all of the modules may be selected according to actual needs to achieve the purpose of the scheme of the present invention. Ordinary technicians in this field can understand and implement it without paying creative work.

[0029] An embodiment of the present invention further provides a computer-readable storage medium having a program stored thereon. When the program is executed by a processor, a field of view leveling device for an ultra-large field of view microscope driven by an electric cylinder in the above embodiment is implemented.

[0030] The computer-readable storage medium may be an internal storage unit of any device with data processing capability described in any of the aforementioned embodiments, such as a hard disk or a memory. The computer-readable storage medium may also be an external storage device of any device with data processing capability, such as a plug-in hard disk, a smart media card (SMC), an SD card, a flash card, etc. equipped on the device. Furthermore, the computer-readable storage medium may also include both an internal storage unit and an external storage device of any device with data processing capability. The computer-readable storage medium is used to store the computer program and other programs and data required by any device with data processing capability, and may also be used to temporarily store data that has been output or is to be output.

[0031] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder, characterized in that: The method comprises an electric cylinder, a lens and a microscope stage, wherein the microscope stage comprises an X-axis and a Y-axis for controlling the deflection of the stage on the X-axis and the Y-axis, and a sample to be photographed is arranged on the microscope stage, and the method comprises the following steps: S1: Capture a microscopic image of the sample at the starting angle through the lens to obtain a starting microscopic image; S2: starting from the starting angle, collecting microscopic images at several angles on the X-axis within a set deflection angle range, and obtaining several deflection microscopic images; S3: After each angle deflection, the deflected microscopic image is registered and cropped according to the starting microscopic image, and the common field of view area is retained to ensure the accuracy of subsequent images; S4: Calculate the clarity of the microscopic images at the starting angle and several angles; S5: Obtain the X-axis angle with the highest definition, return to S2 based on the angle and execute from the Y-axis to obtain the Y-axis angle with the highest definition, and complete the global field of view leveling of the photographed sample in the XY dimensions.

2. The method for leveling the field of view of a super-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: The global field of view leveling in S5 is achieved through iterative optimization, and the order of leveling the X-axis and the Y-axis is interchangeable; S2 includes collecting from any one of the X-axis and the Y-axis, and returning to S2 in S5 is executed from the other one of the X-axis and the Y-axis.

3. The method for leveling the field of view of a super-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: The S1 includes the following contents: obtaining the region of interest of the sample being photographed, and taking the current position as the starting angle, collecting the microscopic image at the current angle.

4. The method for leveling the field of view of a super-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: The S2 comprises the following sub-steps: S21: acquiring a plurality of acquisition angles within a preset deflection angle range according to the starting angle; S22: collecting microscopic images at several collection angles; S23: Acquire an image sequence based on the microscopic images at several angles.

5. The method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: S3 includes the following contents: since the photographed samples in the field of view move during the angle deflection process, each image needs to be translated and registered based on the starting angle as a reference, and the registered images are cropped to retain the common area.

6. A method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder as claimed in claim 5, characterized in that: The translation registration in S3 includes the following contents: registering the microscopic image by using the matchTemplate function in OpenCV.

7. The method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: S4 includes the following contents: for the initial microscopic image and the deflected microscopic image cropped in S3, the total variation is calculated by using the height, width and row and column indexes of the images respectively; under the premise of having the same foreground target, the larger the total variation, the higher the clarity.

8. The method for leveling the field of view of a super-large field of view microscope driven by an electric cylinder as claimed in claim 1, characterized in that: S5 includes the following contents: according to the calculation result of S4, extract the deflection microscopic image corresponding to the clearest image, and deflect the microscope stage to the angle; then return to S2, repeat the contents of S2 to S4 on the Y axis, obtain the clearest angles on the X axis and the Y axis, and then realize the global field of view leveling in the XY two-dimensional.

9. A field of view leveling device for an ultra-large field of view microscope driven by an electric cylinder, characterized in that: It comprises a memory and one or more processors, wherein the memory stores executable codes, and when the one or more processors execute the executable codes, they are used to implement the field of view leveling method for an ultra-large field of view microscope driven by an electric cylinder as described in any one of claims 1 to 8.

10. A computer-readable storage medium, characterized in that: A program is stored thereon, and when the program is executed by a processor, a method for leveling the field of view of an ultra-large field of view microscope driven by an electric cylinder as described in any one of claims 1 to 8 is implemented.

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