An automatic focusing method, apparatus and equipment for a fluorescence microscope

Abstract

An automatic focusing method, apparatus, and device for fluorescence microscopy includes: S10, acquiring a microscope image and extracting the outline of a target cell within the image; S20, identifying the target cell outline to obtain an internal region and an external region, wherein the internal region is the main body of the cell, and the external region is the part outside the main body of the cell; S30, calculating the sharpness values ​​of the internal and external regions separately and performing weighted processing, wherein the sharpness value of the internal region has a greater weight than that of the sharpness value of the external region; S40, calculating a global sharpness score according to a preset formula; and S50, automatically adjusting the microscope focal length based on the global sharpness score. By calculating the sharpness values ​​of the internal and external regions separately and performing weighted processing, priority is given to the detail sharpness of the main body of the cell, effectively reducing the interference of halo effect on the focusing result, and improving focusing accuracy and imaging quality.

Description

Technical Field

[0001] This invention relates to the field of optical microscopy, and in particular to an automatic focusing method, apparatus and equipment for a fluorescence microscope. Background Technology

[0002] In the field of fluorescence microscopy image processing, image sharpness evaluation is crucial for obtaining high-quality imaging results. However, significant halos may occur during fluorescence microscopy imaging, affecting image sharpness and the accuracy of analytical results.

[0003] Current autofocus algorithms mostly rely on image contrast, gradient, or spectral analysis, and tend to perform uniform processing on the entire image. However, when faced with high-halo imaging in fluorescence microscopy, these methods often misjudge halo areas as sharp areas, leading to decreased focusing accuracy. Especially when the halo effect around cells is significant, the algorithm struggles to focus on the main part of the cell, ultimately affecting image quality.

[0004] Therefore, in fluorescence microscopy imaging under defocused conditions, existing automatic focusing methods for fluorescence microscopes are not effective, and halo effects are prone to occur around cells, leading to misjudgments of image sharpness. Summary of the Invention

[0005] The purpose of this invention is to provide an automatic focusing method, apparatus, and device for fluorescence microscopes, aiming to solve the technical problem of poor focusing effect in existing automatic focusing methods for fluorescence microscopes.

[0006] To achieve the above objectives, the present invention provides an automatic focusing method for a fluorescence microscope, comprising the following steps:

[0007] S10. Acquire microscope images and extract the outlines of target cells within the microscope images;

[0008] S20. Identify the outline of the target cell to obtain an inner region and an outer region, wherein the inner region is the main part of the cell and the outer region is the part outside the main part of the cell.

[0009] S30. Calculate the sharpness values ​​of the internal and external regions respectively, and perform weighted processing on them respectively. The weight of the sharpness value of the internal region is greater than the weight of the sharpness value of the external region.

[0010] S40. Calculate the global sharpness score according to the preset formula;

[0011] S50: Automatically adjusts microscope focal length based on global sharpness score.

[0012] Optionally, the automatic adjustment of the microscope focal length based on the global sharpness score in S50 includes at least the following steps:

[0013] S51. Focus the microscope at the preset initial position and set the initial focal length value, then mark it as the current focal length.

[0014] S52. Set the focus range and start focusing from the current focal length minus half of the focus range;

[0015] S53. Execute S10-S40 to obtain the current global sharpness score;

[0016] S54. Determine whether the current global sharpness score is greater than the global sharpness score calculated last time. If yes, execute S55. If no, stop the focus adjustment, automatically record the current focus of the last time, and lock the microscope at the focus position.

[0017] S55. Adjust the focal length with a preset increment, update the current focal length, and execute S52-S54 until the global sharpness score reaches the peak. During the adjustment process, determine in real time whether the stopping condition is met. If it is met, execute S56.

[0018] S56. Automatically record the focal length value when focus adjustment stops, and lock the microscope at that focal point.

[0019] Optionally, the stopping condition is: whether the difference between the current global sharpness score and the previously calculated global sharpness score is less than a threshold for N consecutive times, where N is a preset value;

[0020] Alternatively, is the current global sharpness score half of the peak global sharpness score?

[0021] Optionally, the microscope image is filtered before extracting the target cell outline in the microscope image in S10.

[0022] Optionally, in S10, the outline of the target cell in the microscope image is extracted using an edge detection algorithm or a morphological algorithm.

[0023] Optionally, the preset formula is as follows:

[0024] ;

[0025] Where Double result is the global sharpness score, resultCellInside is the internal region sharpness value, 'a' is the weight corresponding to the internal region sharpness value, resultCellOutside is the external region sharpness value, and 'b' is the weight corresponding to the external region sharpness value.

[0026] Optional, a is 500, b is 1.

[0027] Optionally, in S30, the sharpness values ​​of the internal and external regions are calculated using either the Laplace transform or the variance function.

[0028] Corresponding to the aforementioned automatic focusing method for fluorescence microscopes, the present invention provides an automatic focusing device for fluorescence microscopes, comprising:

[0029] An image processing unit is used to acquire images acquired by a microscope and extract the outlines of target cells within the acquired images.

[0030] The recognition unit is used to recognize the outline of the target cell and obtain an inner region and an outer region, wherein the inner region is the main body of the cell and the outer region is the part outside the main body of the cell;

[0031] The calculation unit is used to calculate the sharpness values ​​of the internal and external regions respectively, and to perform weighted processing on them respectively, wherein the weight of the sharpness value of the internal region is greater than the weight of the sharpness value of the external region; and to calculate the global sharpness score according to a preset formula.

[0032] The focus adjustment unit is used to automatically adjust the microscope focus based on the global sharpness score.

[0033] In addition, to achieve the above objectives, the present invention also provides an autofocus device for a fluorescence microscope, the autofocus device for a fluorescence microscope including a fluorescence microscope and autofocus software stored in the fluorescence microscope, the device being used to implement the autofocus method for a fluorescence microscope as described above.

[0034] The beneficial effects of this invention are:

[0035] (1) Compared with the prior art, the present invention calculates the sharpness values ​​of the internal and external regions separately and performs weighted processing, giving priority to the detail sharpness of the main part of the cell, effectively reducing the interference of the halo effect on the focusing result, and improving the focusing accuracy and imaging quality.

[0036] (2) Compared with the prior art, the present invention automatically adjusts the microscope focal length based on global sharpness score to ensure the systematicness and accuracy of the focusing process, and achieves precise focusing by gradually adjusting the focal length and evaluating sharpness in real time;

[0037] (3) Compared with the prior art, the present invention stops focusing in advance by repeatedly changing the score less than the threshold or reducing the global sharpness score to 1 / 2 of the maximum value of the global sharpness score, thereby optimizing the focusing rate, improving the focusing efficiency, reducing unnecessary focusing adjustments, and saving time and computing resources.

[0038] (4) Compared with the prior art, the present invention reduces noise interference and improves image quality by filtering the microscope images before extracting the target cell outline, thereby improving the accuracy and reliability of subsequent sharpness assessment.

[0039] (5) Compared with the prior art, the present invention uses edge detection algorithm or morphological algorithm to extract the outline of target cells, ensuring the accuracy and robustness of outline extraction, providing a reliable basis for subsequent region marking and sharpness evaluation, and improving the overall performance of the focusing algorithm;

[0040] (6) Compared with the prior art, the present invention highlights the importance of details inside cells by weighting the sharpness values ​​of the internal and external regions, making the focusing algorithm more in line with the user's needs and improving the accuracy of focusing and the image quality.

[0041] (7) Compared with the prior art, the present invention calculates the sharpness values ​​of the internal and external regions through Laplace transform or variance function, providing an effective sharpness evaluation method, ensuring that the focusing algorithm can accurately evaluate the sharpness of the image, and improving the accuracy and reliability of focusing;

[0042] (8) Compared with the prior art, the present invention provides an automatic focusing device for a fluorescence microscope, including an image processing unit, a recognition unit, a calculation unit and a focal length adjustment unit, which realizes the automated process from image acquisition to focus adjustment, improves the efficiency and accuracy of focusing, and is suitable for various fluorescence microscope imaging applications.

[0043] (9) Compared with the prior art, the present invention provides an automatic focusing device for a fluorescence microscope, which integrates a fluorescence microscope and automatic focusing software, realizes the automation and intelligence of the focusing method, improves the ease of use and imaging quality of the microscope, and is suitable for microscope imaging needs in scientific research, medical and industrial fields.

[0044] These or other aspects of this application will become more apparent from the following description of embodiments. It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only, and are not intended to limit the scope of this application. Attached Figure Description

[0045] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other embodiments can be obtained based on these drawings without creative effort.

[0046] In the diagram:

[0047] Figure 1This is a flowchart of an embodiment of the automatic focusing method for a fluorescence microscope according to the present invention.

[0048] Figure 2 This is a frame diagram of an embodiment of the automatic focusing device for a fluorescence microscope according to the present invention. Detailed Implementation

[0049] The present application will now be further described in conjunction with the accompanying drawings and specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments.

[0050] To make the objectives, technical solutions, and advantages of this invention clearer, the embodiments of this invention will be further described in detail below with reference to specific examples and the accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative and are not intended to limit this application.

[0051] It should be noted that all uses of the terms "first" and "second" in the embodiments of the present invention are for the purpose of distinguishing two different entities or different parameters with the same name. Therefore, "first" and "second" are merely for convenience of expression and should not be construed as limiting the embodiments of the present invention. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion, such as other steps or units inherent in a process, method, apparatus, product, or device that includes a series of steps or units.

[0052] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0053] The flowchart shown in the attached diagram is for illustrative purposes only and does not necessarily include all content and operations / steps, nor does it necessarily have to be performed in the described order. For example, some operations / steps can be broken down, combined, or partially merged, so the actual execution order may change depending on the actual situation.

[0054] The following detailed description of some embodiments of this application is provided in conjunction with the accompanying drawings. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0055] See Figure 1 As shown, an embodiment of the present invention provides an automatic focusing method for a fluorescence microscope, comprising the following steps: S10, acquiring a microscope image and extracting the outline of the target cell within the microscope image;

[0056] S20. Identify the outline of the target cell to obtain an inner region and an outer region, wherein the inner region is the main part of the cell and the outer region is the part outside the main part of the cell.

[0057] S30. Calculate the sharpness values ​​of the internal and external regions respectively, and perform weighted processing on them respectively. The weight of the sharpness value of the internal region is greater than the weight of the sharpness value of the external region.

[0058] S40. Calculate the global sharpness score according to the preset formula;

[0059] S50: Automatically adjusts microscope focal length based on global sharpness score.

[0060] Preferably, S20 includes at least the following steps:

[0061] S21. Adaptive threshold segmentation is performed on microscope images using histogram double thresholding methods (including triangle method and Otsu method) to extract bright spots and target contour regions.

[0062] S22. After removing the larger and smaller contours, the remaining contour area is identified as the outer contour.

[0063] S23. The outer contour is structured using operators such as dilation and erosion to obtain the inner and outer regions.

[0064] The outer region is mainly used for sharpness calculation of cell boundary contours, while the inner region is mainly used for sharpness calculation of detailed features within the cell's interior.

[0065] It should be noted that, in specific application scenarios, the standards for "larger" and "smaller" can be set based on the observed cell diameter. For example, if the observed cell diameter is 10µm, then "larger" specifically means greater than 15µm, and "smaller" specifically means less than 5µm. In this application scenario, contours larger than 15µm and smaller than 5µm are removed, while contours with diameters between 5µm and 15µm are retained.

[0066] That is, in this embodiment, the specific standards for "larger" and "smaller" are set according to the diameter of the observed cells. Based on the cell diameter, the larger standard is set to 1.5 times the cell diameter, and the smaller standard is set to 1 / 2 the cell diameter.

[0067] This invention calculates and weights the sharpness values ​​of the internal and external regions separately, prioritizing the detail sharpness of the main cell parts, effectively reducing the interference of halo effect on the focusing results, and improving focusing accuracy and image quality.

[0068] In this embodiment, the automatic adjustment of the microscope focal length based on the global sharpness score in step S50 includes at least the following steps:

[0069] S51. Focus the microscope at the preset initial position and set the initial focal length value, then mark it as the current focal length.

[0070] S52. Set the focus range and start focusing from the current focal length minus half of the focus range;

[0071] S53. Execute S10-S40 to obtain the current global sharpness score;

[0072] S54. Determine whether the current global sharpness score is greater than the global sharpness score calculated last time. If yes, execute S55. If no, stop the focus adjustment, automatically record the current focus of the last time, and lock the microscope at the focus position.

[0073] S55. Adjust the focal length with a preset increment, update the current focal length, and execute S52-S54 until the global sharpness score reaches the peak. During the adjustment process, determine in real time whether the stopping condition is met. If it is met, execute S56.

[0074] S56. Automatically record the focal length value when focus adjustment stops, and lock the microscope at that focal point.

[0075] In this embodiment, in S52, focusing begins at the current focal length - focus range / 2. The focus range is primarily set based on the sample. If the sample is relatively flat, a smaller range can be set; if the sample is uneven, a larger range is required. "Relatively flat sample" means that the image surface being processed during focus adjustment is relatively flat, without significant undulations or complex textures. In this case, the focus range can be set smaller because a smaller focus range is sufficient to cover the entire image or object, ensuring its sharpness. Conversely, if the sample surface is uneven, with significant variations in height or complex textures, then the focus range needs to be set larger to ensure that areas of different depths are correctly focused, thereby achieving better overall sharpness.

[0076] Preferably, the preset increment in S55 can be preset according to actual needs. The preset increment is specifically related to the depth of field of the objective lens, the exposure time during focusing, and the Z-axis movement speed. The smaller the preset increment, the higher the focusing accuracy and the longer the focusing time.

[0077] This invention automatically adjusts the microscope focal length based on a global sharpness score, ensuring the systematic nature and accuracy of the focusing process. By gradually adjusting the focal length and evaluating sharpness in real time, it achieves precise focusing.

[0078] In this embodiment, the stopping condition is specifically: whether the difference between the current global sharpness score and the previously calculated global sharpness score is less than a threshold for N consecutive times, where N is a preset value;

[0079] Alternatively, is the current global sharpness score half of the peak global sharpness score?

[0080] Preferably, the threshold is 1 / 20 of the current global clarity score.

[0081] It should be noted that the peak global sharpness score is specifically the maximum global sharpness score.

[0082] This invention stops focusing early by repeatedly changing the score to less than a threshold or by reducing the global sharpness score to half of the maximum global sharpness score. This optimizes the focusing rate, improves focusing efficiency, reduces unnecessary focusing adjustments, and saves time and computing resources.

[0083] In this embodiment, before extracting the target cell outline in the microscope acquisition image in S10, the microscope acquisition image is also filtered.

[0084] This invention improves the accuracy and reliability of subsequent sharpness assessment by filtering microscope images before extracting the target cell outline, thereby reducing noise interference and improving image quality.

[0085] In this embodiment, in S10, the outline of the target cell in the microscope image is extracted by an edge detection algorithm or a morphological algorithm.

[0086] This invention employs edge detection or morphological algorithms to extract the contours of target cells, ensuring the accuracy and robustness of contour extraction. This provides a reliable foundation for subsequent region marking and sharpness assessment, thereby improving the overall performance of the focusing algorithm.

[0087] In this embodiment, the preset formula is as follows:

[0088] ;

[0089] Where Double result is the global sharpness score, resultCellInside is the internal region sharpness value, 'a' is the weight corresponding to the internal region sharpness value, resultCellOutside is the external region sharpness value, and 'b' is the weight corresponding to the external region sharpness value.

[0090] In this embodiment, a is 500 and b is 1.

[0091] It should be noted that the weights 'a' for the internal region sharpness value and 'b' for the external region sharpness value are not fixed at 500 and 1. If the sample differences are significant (judged according to actual needs), they can be adjusted accordingly. The range of values ​​for 'a' and 'b' is [1, 10000]. The principle for adjusting according to actual needs is to assign a higher weight to the details and textures inside the cells (i.e., 'a' > 'b'). By adjusting the weights 'a' for the internal region sharpness value and 'b' for the external region sharpness value, focus evaluation can be dynamically optimized according to different sample types, resulting in higher adaptability.

[0092] This invention emphasizes the importance of details inside cells by weighting the sharpness values ​​of internal and external regions, making the focusing algorithm more in line with user needs and improving focusing accuracy and image quality.

[0093] In this embodiment, in S30, the sharpness values ​​of the internal and external regions are calculated by Laplace transform or variance function, respectively.

[0094] This invention calculates the sharpness values ​​of the internal and external regions using Laplace transform or variance function, providing an effective sharpness evaluation method. This ensures that the focusing algorithm can accurately evaluate the sharpness of the image, improving the accuracy and reliability of focusing.

[0095] like Figure 2 As shown, the present invention also provides an automatic focusing device for a fluorescence microscope, comprising:

[0096] Image processing unit 10 is used to acquire microscope images and extract the outline of target cells within the microscope images;

[0097] The recognition unit 20 is used to recognize the outline of the target cell and obtain an inner region and an outer region, wherein the inner region is the main body of the cell and the outer region is the part outside the main body of the cell.

[0098] The calculation unit 30 is used to calculate the sharpness values ​​of the internal region and the external region respectively, and to perform weighted processing on them respectively, wherein the weight corresponding to the sharpness value of the internal region is greater than the weight corresponding to the sharpness value of the external region; and to calculate the global sharpness score according to a preset formula.

[0099] The focal length adjustment unit 40 is used to automatically adjust the microscope focal length based on the global sharpness score.

[0100] This invention provides an automatic focusing device for a fluorescence microscope, comprising an image processing unit 10, a recognition unit 20, a calculation unit 30, and a focal length adjustment unit 40. It realizes an automated process from image acquisition to focus adjustment, improves focusing efficiency and accuracy, and is suitable for various fluorescence microscope imaging applications.

[0101] This invention also provides an autofocus device for a fluorescence microscope, which includes a fluorescence microscope and autofocus software stored in the fluorescence microscope. The device is used to implement the autofocus method for a fluorescence microscope as described above.

[0102] This invention provides an automatic focusing device for a fluorescence microscope, which integrates a fluorescence microscope and automatic focusing software, realizing the automation and intelligence of the focusing method, improving the ease of use and imaging quality of the microscope, and is suitable for microscope imaging needs in scientific research, medical and industrial fields.

[0103] It should be noted that the various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. Similar or identical parts between embodiments can be referred to interchangeably. For the apparatus and device embodiments, since they are basically similar to the method embodiments, the descriptions are relatively simple; relevant parts can be referred to the descriptions in the method embodiments.

[0104] The foregoing description illustrates and describes preferred embodiments of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the inventive concept by means of the foregoing teachings or techniques or knowledge in related fields. Any modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be protected within the scope of the appended claims.

Claims

1. A method for automatic focusing of a fluorescence microscope, characterized in that, Includes the following steps: S10. Acquire microscope images and extract the outlines of target cells within the microscope images; S20. Identify the outline of the target cell to obtain an inner region and an outer region, wherein the inner region is the main body of the cell and the outer region is the part outside the main body of the cell, including at least the following steps: S21. Adaptive threshold segmentation is performed on microscope images using the histogram double threshold method to extract bright spots and target contour regions; the histogram double threshold method includes the triangle method and the Otsu method. S22. After removing the larger and smaller contours, the remaining contour region is identified as the outer contour. The outer contour is structured using the dilation operator to obtain the inner region. The outer contour is then structured using operators such as erosion to obtain the outer region. The criteria for the larger and smaller contours can be set according to the observed cell diameter. S30. Calculate the sharpness values ​​of the internal and external regions respectively, and perform weighted processing on them respectively. The weight of the sharpness value of the internal region is greater than the weight of the sharpness value of the external region. S40. Calculate the global sharpness score according to the preset formula; S50. Automatically adjust the microscope focal length based on the global sharpness score, including at least the following steps: S51. Focus the microscope at the preset initial position and set the initial focal length value, marking it as the current focal length; S52. Set the focus range and start focusing from the current focal length minus half of the focus range; S53. Execute S10-S40 to obtain the current global sharpness score; S54. Determine whether the current global sharpness score is greater than the global sharpness score calculated last time. If yes, execute S55. If no, stop the focus adjustment, automatically record the current focus of the last time, and lock the microscope at the focal position corresponding to the recorded current focus of the last time. S55. Adjust the focal length with a preset increment, update the current focal length, and execute S52-S54 until the global sharpness score reaches the peak. During the adjustment process, determine in real time whether the stopping condition is met. If it is met, execute S56. S56. Stop focusing, automatically record the current focal length, and lock the microscope at the focal point corresponding to the recorded current focal length.

2. The automatic focusing method for a fluorescence microscope according to claim 1, characterized in that: The stopping condition is as follows: whether the difference between the current global sharpness score and the previously calculated global sharpness score is less than a threshold for N consecutive times, where N is a preset value; Alternatively, is the current global sharpness score half of the peak global sharpness score? 3. The automatic focusing method for a fluorescence microscope according to claim 1, characterized in that: Before extracting the target cell outline within the microscope acquisition image, the microscope acquisition image is filtered in S10.

4. The automatic focusing method for a fluorescence microscope according to claim 1, characterized in that: In S10, the outline of target cells within the microscope-acquired image is extracted using edge detection or morphological algorithms.

5. The automatic focusing method for a fluorescence microscope according to claim 1, characterized in that: The preset formula is as follows: ； Where Double result is the global sharpness score, resultCellInside is the internal region sharpness value, 'a' is the weight corresponding to the internal region sharpness value, resultCellOutside is the external region sharpness value, and 'b' is the weight corresponding to the external region sharpness value.

6. The automatic focusing method for a fluorescence microscope according to claim 5, characterized in that: a is 500, b is 1.

7. The automatic focusing method for a fluorescence microscope according to claim 1, characterized in that: In S30, the sharpness values ​​of the internal and external regions are calculated using either the Laplace transform or the variance function.

8. An automatic focusing device for a fluorescence microscope, characterized in that, include: An image processing unit is used to acquire images acquired by a microscope and extract the outlines of target cells within the acquired images. The recognition unit is used to identify the contour of the target cell, obtaining an internal region and an external region. The internal region is the main body of the cell, and the external region is the part outside the main body of the cell. The unit includes at least the following steps: S21, adaptive thresholding of the microscope image using a histogram double thresholding method to extract bright spots and target contour regions; wherein the histogram double thresholding method includes the triangle method and the Otsu method; S22, after removing larger and smaller contours, the remaining contour region is identified as the external contour; the external contour is structured using a dilation operator to obtain the internal region; the external contour is structured using operators such as erosion to obtain the external region; wherein the criteria for larger and smaller contours can be set according to the observed cell diameter. The calculation unit is used to calculate the sharpness values ​​of the internal and external regions respectively, and to perform weighted processing on them respectively, wherein the weight of the sharpness value of the internal region is greater than the weight of the sharpness value of the external region; and to calculate the global sharpness score according to a preset formula. The focal length adjustment unit is used to automatically adjust the microscope focal length based on a global sharpness score, and includes at least the following steps: S51, focusing the microscope at a preset initial position and setting an initial focal length value, which is then marked as the current focal length; S52, setting the focus range and starting focusing from half of the current focal length minus the focus range; S53, executing the corresponding functions of the image processing unit, recognition unit, calculation unit, and focal length adjustment unit to obtain the current global sharpness score; S54, determining whether the current global sharpness score is greater than the previously calculated global sharpness score. If the score is positive, proceed to step S55; otherwise, stop focusing, automatically record the previous current focus, and lock the microscope at the focal position corresponding to the recorded previous current focus. Step S55: Adjust the focus using a preset increment, update the current focus, and proceed to steps S52-S54 until the global sharpness score reaches its peak. During the adjustment process, continuously check if the stopping condition is met. If it is, proceed to step S56. Step S56: Stop focusing, automatically record the current focus, and lock the microscope at the focal position corresponding to the recorded current focus.

9. An automatic focusing device for a fluorescence microscope, characterized in that, The fluorescence microscope autofocus device includes a fluorescence microscope and autofocus software stored in the fluorescence microscope, the device being used to implement the fluorescence microscope autofocus method as described in any one of claims 1 to 7.

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