A high-speed microscopic image acquisition method

Abstract

The application provides a high-speed microscopic image acquisition method, and a microscopic image acquisition device including a stepping stage, a microscope tube, an industrial camera, a stroboscopic head and an industrial computer; the stepping stage is used for loading a slide, and an x-axis stepping motor and a y-axis stepping motor are arranged on the stepping stage; a stroboscopic pulse signal is sent to the stroboscopic head; an exposure pulse signal is synchronously sent to the industrial camera; the industrial camera is exposed to acquire current field image data at the same time when the stroboscopic head flashes; the image data is stored and spliced according to coordinates; and the above steps are used to realize high-speed acquisition of microscopic images. The image acquisition scheme based on continuous scanning of a rectangular field is adopted, and the stroboscopic head is used to improve the image acquisition illumination, so that the image acquisition efficiency is greatly improved under the premise of ensuring the image acquisition precision, and the image acquisition time is reduced to 60-120 seconds.

Description

Technical Field

[0001] This invention relates to the field of microscopic image acquisition, and in particular to a high-speed microscopic image acquisition method. Background Technology

[0002] Cellular and tissue pathology image recognition technology is considered an authoritative and definitive diagnostic method. However, the detection process for cellular and tissue pathology image recognition technology is lengthy, including sampling, slide preparation, sample microscopic image acquisition, image stitching, and image recognition. In the era of entirely manual labor, a pathologist could only complete a single-digit number of diagnoses per day. This is far from meeting user needs. A bottleneck in the existing diagnostic process is the high cost and time-consuming nature of microscopic image scanning equipment. Internationally, Dr. Weinstein, in collaboration with the US-based D Metrix research and development team, developed the DX-40 rapid digital slide microscopy scanner, a pathology diagnostic product. It integrates 80 microscopes for simultaneous scanning, achieving a processing speed of 40 images per hour. However, its complex structure results in a high price. The applicant has previously developed a series of miniature microscopic image acquisition devices, such as CN110794569A, a miniature cell microscopic image acquisition device and image recognition method. Although using AI-based stitching and recognition, acquiring and processing a single slide image with this device still takes approximately two hours. The main technical bottleneck lies in the intermittent image acquisition method, requiring a stop position at each field of view to allow for lens exposure. Acquiring a 1200-field slide takes about 20 minutes, resulting in low efficiency. With the increasing prevalence of cell and tissue pathology image recognition technology, the efficiency of existing equipment limits its development. US Patent 8755579A describes a fully automated, rapid microscope slide scanner that employs a continuous line scanning and real-time stitching scheme, significantly improving acquisition speed. However, this approach demands extremely high equipment precision; otherwise, the stitched microscopic images will have significant errors and lack diagnostic value. Furthermore, there are few dedicated lenses available for linear scanners on the market, leading to an exponential increase in both image quality and cost. Summary of the Invention

[0003] The technical problem this invention aims to solve is to provide a high-speed microscopic image acquisition method that can significantly improve the speed of image acquisition, utilizes largely mature components, and ensures the quality of scanned images. It can also correct the position or imaging quality of the acquired images, reducing the requirements for high-precision components.

[0004] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is: a high-speed microscopic image acquisition method, wherein the microscopic image acquisition device includes a stepping stage, a microscope tube, an industrial camera, a stroboscopic head, and an industrial control computer;

[0005] The stepper stage is used to load glass slides. The stepper stage is equipped with an x-axis stepper motor and a y-axis stepper motor.

[0006] The steps of the control method are as follows:

[0007] S1. Calculate the number of step pulse signals based on the field of view;

[0008] S2. Within the field of view, the sum of the periods of the step pulse signals is taken as the strobe period;

[0009] S3. Continuously send stepping pulse signals to the stepper motor;

[0010] S4. The first field of view takes 1 / 2 of the strobe cycle and sends a strobe pulse signal to the strobe head;

[0011] Simultaneously send exposure pulse signals to the industrial camera;

[0012] While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view;

[0013] S5. The subsequent field of view sends a strobe pulse signal to the strobe head according to the strobe cycle;

[0014] Simultaneously send exposure pulse signals to the industrial camera;

[0015] While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view;

[0016] S6. Store image data and stitch it together according to coordinates;

[0017] The above steps enable high-speed acquisition of microscopic images.

[0018] In the preferred embodiment, in step S1, the field of view refers to the physical size corresponding to the single-exposure image data on the slide, including the length in the x-direction and the length in the y-direction;

[0019] The number of step pulse signals in the x-direction length ≤ x-direction field of view length / ((single step pulse rotation angle / 360) × x-axis screw pitch);

[0020] The number of step pulse signals in the y-direction length ≤ y-direction field of view length / ((single step pulse rotation angle / 360) × y-axis screw pitch);

[0021] In the preferred embodiment, in step S2, the strobe period includes an x-axis strobe period and a y-axis strobe period. The x-axis strobe period is the sum of the periods of the step pulse signals of x-axis length, and the y-axis strobe period is the sum of the periods of the step pulse signals of y-axis length.

[0022] In the preferred embodiment, in step S3, the scanning path of the stepping stage is "S" shaped, that is, the slide moves one row along the x-direction relative to the microscope tube, then moves to the next row along the y-direction, and then traverses the current row again, and so on to complete the acquisition of slide image data.

[0023] In the preferred embodiment, the width of the strobe pulse signal is greater than the width of the exposure pulse signal;

[0024] That is, the time for the stroboscopic light to shine is longer than the exposure time.

[0025] In the preferred embodiment, the illuminance of the stroboscopic head is 50,000 lx to 100,000 lx, the stepping speed of the stepping stage is 17 to 20 ms / field of view, the numerical aperture of the microscope tube is 0.5 to 0.75, and the exposure time of the industrial camera is 10 to 25 microseconds.

[0026] In a preferred embodiment, the structure of the strobe head is such that the LED light source is electrically connected to the output terminal of the switching element, the control terminal of the switching element is electrically connected to the controller, and the input terminal of the switching element is electrically connected to the power supply.

[0027] In the preferred embodiment, an x-axis travel sensor is provided on the stepping stage to verify the relative displacement between the base and the x-axis seat. If an error is detected, the error value is sent and stored as the x-axis correction value for subsequent image stitching.

[0028] The stepper stage is equipped with a y-axis travel sensor to verify the relative displacement between the x-axis and y-axis seats. If an error is detected, the error value is sent and stored as the y-axis correction value for subsequent image stitching.

[0029] In the preferred embodiment, step S6 includes the following steps during splicing: S61, splicing the preset field of view frame into a complete field of view;

[0030] Convert the field-of-view image to its actual size in pixels;

[0031] S62. In the first two fields of view, the first field of view image is filled into the field of view frame in a centered alignment manner. The second field of view image is filled into the second field of view frame in the x-direction with the first field of view image as a reference and with an overlay alignment operation. The x-direction offset value between the center of the field of view image and the field of view frame is stored as the x-direction filling correction parameter for the subsequent field of view image.

[0032] S63. Assign the x-direction correction value to each corresponding field of view, sum the filling correction parameter with the current x-direction correction value, and obtain the filling parameter in the back field of view image.

[0033] S64. When a new line is inserted, the preceding field of view image is used as a reference, and the field of view frame in the following line is filled in the y direction using an overlay alignment operation. The y-direction offset value between the center of the field of view image and the field of view frame is stored as the y-direction filling correction parameter for the subsequent field of view image.

[0034] S65. Assign the y-direction correction value to the corresponding field of view for each line break, and sum the fill correction parameter with the current x-direction correction value to obtain the fill parameter of the field of view image after the line break.

[0035] The above steps enable rapid stitching of visual field images.

[0036] In a preferred embodiment, a light compensation sensor is also provided. In steps S4 and S5, the illuminance value of the stroboscopic flash collected by the light compensation sensor is synchronously acquired according to the exposure pulse signal. In step S6, the illuminance value of the stroboscopic flash corresponding to each field of view is averaged to obtain the difference between the illuminance value of the stroboscopic flash in each field of view and the average value. This difference is compared with a preset illuminance value error range. If the difference exceeds the range, the brightness value of the current field of view image is compensated and adjusted.

[0037] The high-speed microscopic image acquisition method provided by this invention, by adopting the above-described technical solution, has the following beneficial effects compared with the prior art:

[0038] 1. This invention employs an image acquisition scheme based on continuous scanning of a rectangular field of view, combined with a strobe head to improve image acquisition illumination. While ensuring image acquisition accuracy, it significantly improves image acquisition efficiency, reducing the acquisition time from over 20 minutes to 60-120 seconds (depending on the number of fields of view). For example... Figure 8 As shown, the image with 1392 fields of view, or 29×48 fields of view, was scanned and stitched in just 71 seconds, reaching the international leading level.

[0039] 2. By adopting a continuous walking + stroboscopic approach, a smaller aperture value can be used in the microscope tube, resulting in a deeper field of view for the acquired images, improved diffraction-limited image accuracy, and less image edge distortion and dispersion.

[0040] 4. By using a strobe method, the heat dissipation pressure of the lighting source is reduced due to the shorter working time, allowing for the use of higher-power LED light sources. The preferred working illuminance of the LED light source of this invention is [value missing].

[0041] 5. An open-loop control scheme combined with post-processing error correction was adopted, which further improved the acquisition speed while ensuring splicing accuracy.

[0042] 6. The light compensation sensor is set up so that the illuminance can be unified between different fields of view through illuminance compensation in the later stage, which overcomes the technical problem of uneven brightness values ​​of different fields of view in the flickering mode.

[0043] 7. The method of the present invention can use mature components of existing technology, and through precise control and simplified algorithms, it can significantly reduce equipment costs, making it easier for microscopic scanners to be promoted in primary medical institutions, so as to benefit more users, and intercept malignant tumors such as cervical cancer in the initial stage, thereby greatly improving the cure rate of tumors. Attached Figure Description

[0044] The present invention will be further described below with reference to the accompanying drawings and embodiments:

[0045] Figure 1 This is a flowchart of the present invention.

[0046] Figure 2 This is a schematic diagram of the synchronization control pulse of the present invention.

[0047] Figure 3 This is a schematic diagram of the overall structure of the present invention.

[0048] Figure 4 This is a perspective view of the stage support of the present invention.

[0049] Figure 5 This is a schematic diagram of the field of view, field of view frame, and path of the present invention.

[0050] Figure 6 This is a schematic diagram of the structure of the strobe head of the present invention.

[0051] Figure 7 This is a schematic diagram of the glass slide structure of the present invention.

[0052] Figure 8 This is a schematic diagram of the scanning result interface of the method of the present invention.

[0053] In the diagram: 1. Display device; 2. Industrial computer; 3. Controller; 4. Memory; 5. Z-axis mount; 6. Z-axis stepper motor; 7. Z-axis screw; 8. Z-axis slide; 9. Microscope tube mount; 10. Light compensation sensor; 11. Panoramic lens; 12. Slide holder; 13. Y-axis mount; 14. X-axis mount; 15. X-axis arm; 16. Y-axis motor mount; 17. Y-axis stepper motor; 18. Y-axis screw; 19. Y-axis arm; 20. Y-axis travel sensor; 21. X-axis motor mount; 22. X-axis travel sensor; 23. X-axis stepper motor; 24. X-axis screw; 25. Base; 26. Switching element; 27. Switching control signal; 28. Power supply; 29. ​​Encoded image; 30. Slide; 31. Tissue sample; 32. Industrial camera; 33. Strobe head; 34. LED light source; 35. Strobe period; 36. Field of view 100; Scanning path 200; Field of view frame 300. Detailed Implementation

[0054] like Figure 3 , 4A high-speed microscopic image acquisition method, wherein the microscopic image acquisition device includes a stepping stage, a microscope tube 10, an industrial camera 33, a stroboscopic head 34, and an industrial control computer 2;

[0055] The microscope tube 10 is connected to the industrial camera 33, and the optical elements in the microscope tube 10 are existing technology. The microscope tube 10 is fixedly connected to the z-axis mount 5 of the stepping stage; a strobe head 34 is also provided coaxially or on one side of the microscope tube 10. The strobe head 34 is used to emit flashes at a set frequency according to the continuous travel of the stepping stage, and the industrial camera 33 follows the flash frequency of the strobe head 34 to acquire images. A preferred embodiment is as follows: Figure 6 In the above, the structure of the strobe head 34 is as follows: the LED light source 35 is electrically connected to the output terminal of the switching element 27, the control terminal of the switching element 27 is electrically connected to the controller, and the input terminal of the switching element 27 is electrically connected to the power supply 29; in this example, the switching element 27 is a PMOS transistor.

[0056] The stroboscope 34 has a working illumination of 50,000 lx to 100,000 lx, a stepping stage with a stepping speed of 17 to 20 ms / field of view, a numerical aperture of 0.5 to 0.75 for the microscope tube 10, and an exposure time of 10 to 25 microseconds for the industrial camera 33. The flash duration of the stroboscope 34 is 30 to 120 microseconds. The flash duration covers the exposure time.

[0057] like Figure 3 , 4 In the middle, the structure of the stepper stage is as follows: the base 26 is fixedly connected to the z-axis seat 5, the top of the base 26 is provided with an x-axis groove, the x-axis seat 15 is set in the x-axis groove and slides along the x-axis, the base 26 is provided with an x-axis motor seat 22 extending to the y-axis side, the x-axis stepper motor 24 is fixedly connected to the x-axis motor seat 22, the x-axis arm 16 is provided on the corresponding side of the x-axis seat 15, the x-axis arm 16 is fixedly provided with a nut, the x-axis stepper motor 24 is fixedly connected to the x-axis screw 25, and the x-axis screw 25 is threadedly connected to the nut;

[0058] A groove in the y direction is provided on the top of the x-direction seat 15. The y-direction seat 14 is set in the groove in the y direction and slides along the y direction. A y-axis motor seat 17 extending to the x-direction side is provided on the x-direction seat 15. The y-axis motor seat 17 is fixedly connected to the y-axis stepper motor 18. An extended y-arm 20 is provided on the corresponding side of the y-direction seat 14. A nut is fixed on the y-arm 20. The y-axis stepper motor 18 is fixedly connected to the y-axis screw 19. The y-axis screw 19 is threadedly connected to the nut.

[0059] The top of the y-axis seat 14 is provided with a groove that opens outwards, and the slide holder 13 is disposed in the groove.

[0060] The x-axis stepper motor 24 and the y-axis stepper motor 18 are connected to the controller 3, preferably an ATM32F series controller, which employs open-loop control to achieve higher operating efficiency. Figure 5 , 8 As shown, using the method of this invention, the field of view 100 is a rectangular field of view with a 16:9 aspect ratio, completing 1392 fields of view, i.e., a 29×48 array scan, in 71 seconds. Furthermore, the panoramic image is stitched together immediately after the scan, significantly improving efficiency compared to existing technologies. An artificial intelligence image recognition program is integrated into the industrial control computer 2. This program identifies the range of the tissue sample 32 using the panoramic image and optimizes the scanning path during subsequent scans, removing unnecessary fields of view without content in advance, further improving scanning speed.

[0061] The stepping stage is used to load glass slides 31. The top of the stepping stage has a groove for mounting glass slide holders 13. Typically, each glass slide holder 13 holds two glass slides 31.

[0062] Preferred solutions include Figure 3 The microscope tube 10 is also equipped with a light compensation sensor 11, preferably mounted on the microscope tube 10.

[0063] like Figure 1 In this context, the control method involves the following steps:

[0064] S1. Calculate the number of step pulse signals based on the field of view of 100;

[0065] In the preferred embodiment, in this step, the field of view 100 refers to the physical size corresponding to the single-exposure image data on the slide 31, including the x-axis length and the y-axis length;

[0066] The number of step pulse signals in the x-direction length ≤ x-direction field of view length / ((single step pulse rotation angle / 360) × x-axis screw pitch);

[0067] The number of step pulse signals in the y-direction length ≤ y-direction field of view length / ((single step pulse rotation angle / 360) × y-axis screw pitch);

[0068] This step yields the travel distance of a single stepping pulse signal, and also the relative displacement between the slide 31 and the microscope tube 10. It should be noted that within a field of view, i.e., within a single exposure, the total travel distance of the slide 31 driven by all stepping pulse signals must be less than the physical size corresponding to the field of view image. This is necessary to ensure that adjacent field of view images overlap and to remove blurred edges from the field of view images.

[0069] S2. Within the field of view, the sum of the periods of the step pulse signals is taken as the strobe period 36; in a preferred embodiment, the strobe period 36 includes the x-direction strobe period and the y-direction strobe period. The x-direction strobe period is the sum of the periods of the step pulse signals of the x-direction length, and the y-direction strobe period is the sum of the periods of the step pulse signals of the y-direction length.

[0070] like Figure 2 As shown, for ease of observation, Figure 2 The pulse signal in the image has been simplified. The strobe period 36 is the sum of the step pulse signal times corresponding to the entire distance traveled within the field of view. For example, if the walking time for a distance of 100° field of view is 19ms, then the strobe period 36 time is 19ms.

[0071] S3. Continuously send stepping pulse signals to the stepper motors; the stepping pulse signals are issued by the controller 3 according to the instructions of the industrial computer 2. The x-axis stepper motor 24 and the y-axis stepper motor 18 rotate by one angle according to the stepping pulse signals to achieve precise control of the stroke.

[0072] Preferred solutions include Figure 5 In the process, the scanning path 200 of the stepping stage is "S" shaped, that is, the slide 31 moves one row along the x-direction relative to the microscope tube 10, then moves to the next row along the y-direction, and then traverses the current row again, and so on to complete the acquisition of image data of the slide 31.

[0073] S4. The first field of view is 100, and half of the strobe cycle (36) is used to send a strobe pulse signal to the strobe head; the strobe head's illumination duration is 30~60 microseconds;

[0074] The exposure pulse signal is synchronously sent to the industrial camera; preferably, the exposure time is 10~25 microseconds, and the exposure duration of the stroboscopic head is longer than the exposure time. This is because the stroboscopic head needs both a rising edge and a falling edge to reach its peak value. Therefore, the peak position needs to be aligned with the exposure time. Consequently, the width of the stroboscopic pulse signal is greater than the width of the exposure pulse signal, and the two are centered and aligned. That is, the stroboscopic pulse signal begins to rise earlier than the exposure pulse signal and begins to fall later than the exposure pulse signal.

[0075] In the preferred embodiment, the illuminance of the stroboscopic head 34 is 50,000 lx to 100,000 lx, and the preferred working illuminance is 100,000 lx. The stepping speed of the stepping stage is 17 to 20 ms / field of view, the numerical aperture of the microscope tube 10 is 0.5 to 0.75, and the exposure time of the industrial camera 33 is 10 to 25 microseconds.

[0076] While the strobe head flashes, the industrial camera exposes and acquires 100 image data of the current field of view;

[0077] In the preferred embodiment, the illuminance value of the stroboscopic flash collected by the light compensation sensor 11 is synchronously acquired based on the exposure pulse signal.

[0078] S5. The subsequent field of view 100 sends a strobe pulse signal to the strobe head according to a strobe cycle of 36;

[0079] Simultaneously send exposure pulse signals to the industrial camera;

[0080] While the strobe head flashes, the industrial camera exposes and acquires 100 image data of the current field of view;

[0081] In the preferred embodiment, the illuminance value of the stroboscopic flash collected by the light compensation sensor 11 is synchronously acquired based on the exposure pulse signal.

[0082] S6. Store image data and stitch it together according to coordinates;

[0083] Preferred solutions include Figure 3 In the process, an x-axis travel sensor 23 is provided between the x-axis motor base 22 and the x-arm 16 of the stepping stage. It is used to verify the relative displacement between the base 26 and the x-axis seat 15. If an error is detected, the error value is sent and stored as the x-axis correction value for subsequent image stitching.

[0084] A y-axis travel sensor 21 is provided between the y-axis motor base 17 and the y-arm 20 of the stepper stage. It is used to verify the relative displacement between the x-axis base 15 and the y-axis base 14. If an error is detected, the error value is sent and stored as the y-axis correction value for subsequent image stitching.

[0085] Before stitching, the illuminance values ​​of the stroboscopic lights corresponding to each field of view 100 are averaged to obtain the difference between the illuminance value of each field of view and the average value. This difference is then compared with the preset illuminance value error range. If the difference exceeds the range, the brightness value of the current field of view image is adjusted to compensate.

[0086] In the preferred embodiment, the splicing process includes the following steps:

[0087] S61. Stitch together the preset field of view frame 300 to form a complete field of view;

[0088] The field-of-view image is converted to its actual size in pixels. During calibration, a single field-of-view image from each industrial camera 33 is measured to obtain the physical size corresponding to each pixel—that is, the physical size of a single pixel and the physical size of the field-of-view image. This method enables half-pixel-based stitching during the stitching process, based on floating-point arithmetic precision, resulting in higher stitching accuracy.

[0089] S62, in the first row of two views of 100, such as Figure 5The first and second fields of view, starting from the left in the top row, are filled into the field of view frame 300 with the first field of view image being center-aligned. Center alignment means aligning the field of view image with the midpoint of the field of view frame 300, i.e., centering both vertically and horizontally. The size of the field of view image is larger than the size of the field of view frame 300, and the excess part is clipped by the field of view frame 300 after filling. The subsequent field of view image is filled into the subsequent field of view frame 300 with the first field of view image as a reference, using a superposition alignment operation in the x-direction. The superposition alignment operation calculates the pixels with the same or similar edges of the field of view images to obtain the superposition parameters of the two fields of view. This algorithm is existing technology, such as the stitching method described in CN110879999B, "Microscopic Microscopic Image Acquisition Device and Image Stitching and Recognition Method Based on Mobile Phone". The x-direction offset value between the center of the field of view image and the field of view frame 300 is stored as the x-direction filling correction parameter for the subsequent field of view image.

[0090] S63. Assign the x-direction correction value to each corresponding field of view 100, and sum the filling correction parameter with the current x-direction correction value to obtain the filling parameter in the rear field of view image; the current x-direction correction value refers to the x-direction correction value being assigned to each corresponding field of view 100, so each corresponding field of view 100 needs to be calculated once.

[0091] S64. When a new line is inserted, the preceding field of view image is used as a reference, and the image is filled into the field of view frame 300 in the following line by superimposition and alignment operation in the y direction. The y-direction offset value between the center of the field of view image and the field of view frame 300 is stored as the y-direction filling correction parameter for the subsequent field of view image.

[0092] S65. Assign the y-direction correction value to the corresponding field of view 100 for each line break, sum the fill correction parameter with the current x-direction correction value to obtain the fill parameter of the field of view image after the line break; realize the fast stitching of the field of view image.

[0093] The above steps enable high-speed acquisition of microscopic images.

[0094] The above embodiments are merely preferred technical solutions of the present invention and should not be considered as limitations on the present invention. The embodiments and features described in these embodiments can be arbitrarily combined without conflict. The scope of protection of the present invention should be limited to the technical solutions described in the claims, including equivalent substitutions of the technical features described in the claims. That is, equivalent substitutions and improvements within this scope are also within the scope of protection of the present invention.

Claims

1. A high-speed microscopic image acquisition method, characterized by: The microscopic image acquisition device includes a stepping stage, a microscope tube (10), an industrial camera (33), a strobe head (34), and an industrial control computer (2); The illuminance of the stroboscopic head (34) is 50,000 lx to 100,000 lx, the stepping speed of the stepping stage is 17 to 20 ms / field of view, the numerical aperture of the microscope tube (10) is 0.5 to 0.75, and the exposure time of the industrial camera (33) is 10 to 25 microseconds. The steps of the control method are as follows: S1. Calculate the number of step pulse signals based on the field of view (100); In step S1, the range of the field of view (100) refers to the physical size corresponding to the single exposure image data on the slide (31), including the length in the x direction and the length in the y direction; The number of step pulse signals in the x-direction length ≤ x-direction field of view length / ((single step pulse rotation angle / 360) × x-axis screw pitch); The number of step pulse signals in the y-direction length ≤ y-direction field of view length / ((single step pulse rotation angle / 360) × y-axis screw pitch); The stepper stage is used to load glass slides (31), and the stepper stage is equipped with an x-axis stepper motor (24) and a y-axis stepper motor (18). S2. Take the sum of the periods of the step pulse signals within the field of view as the strobe period (36). In step S2, the strobe period (36) includes the x-direction strobe period and the y-direction strobe period. The x-direction strobe period is the sum of the periods of the step pulse signals of the x-direction length, and the y-direction strobe period is the sum of the periods of the step pulse signals of the y-direction length. S3. Continuously send stepping pulse signals to the stepper motor; S4. The first field of view (100) takes 1 / 2 of the strobe cycle (36) and sends a strobe pulse signal to the strobe head (34). Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head (34) flashes, the industrial camera exposes and acquires image data of the current field of view (100); S5, The subsequent field of view (100) sends a strobe pulse signal to the strobe head (34) according to the strobe cycle (36); Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head (34) flashes, the industrial camera exposes and acquires image data of the current field of view (100); A light compensation sensor (11) is also provided. In steps S4 and S5, the illuminance value of the stroboscopic flash collected by the light compensation sensor (11) is synchronously acquired according to the exposure pulse signal. In step S6, the illuminance value of the stroboscopic flash corresponding to each field of view (100) is averaged to obtain the difference between the illuminance value of the stroboscopic flash of each field of view and the average value. The difference is compared with the preset illuminance value error range. If it exceeds the range, the brightness value of the current field of view image is compensated and adjusted. S6. Store image data and stitch it together according to coordinates; In step S6, the splicing process includes the following steps: S61, splicing the preset field of view frame (300) into a complete field of view; Convert the field-of-view image to its actual size in pixels; S62. In the first two fields of view (100), the first field of view image is filled to the field of view frame (300) in a centered alignment manner. The second field of view image is filled to the second field of view frame (300) in the x direction with an overlay alignment operation based on the first field of view image. The x-direction offset value between the center of the field of view image and the field of view frame (300) is stored as the x-direction filling correction parameter for the subsequent field of view image. S63. Assign the x-direction correction value to each corresponding field of view (100), sum the filling correction parameter with the current x-direction correction value, and obtain the filling parameter in the back field of view image; S64. When a new line is inserted, the first field of view image in the preceding line is used as a reference, and the field of view frame (300) in the following line is filled in the y direction by superposition alignment operation. The y-direction offset value between the center of the field of view image and the field of view frame (300) is stored as the y-direction filling correction parameter of the subsequent field of view image. S65. Assign the y-direction correction value to the corresponding field of view (100) for each line break, and sum the fill correction parameter with the current x-direction correction value to obtain the fill parameter of the field of view image after the line break. The above steps enable high-speed acquisition and stitching of microscopic images.

2. The high-speed microscopic image acquisition method according to claim 1, characterized in that: In step S3, the scanning path (200) of the stepping stage is "S" shaped, that is, the slide (31) moves one row relative to the microscope tube (10) along the x direction, then moves to the next row along the y direction, and then traverses the current row again, and so on to complete the acquisition of slide (31) image data.

3. The high-speed microscopic image acquisition method according to claim 1, characterized in that: The width of the strobe pulse signal is greater than the width of the exposure pulse signal; That is, the time for the stroboscopic head (34) to emit light is longer than the exposure time.

4. The high-speed microscopic image acquisition method according to claim 1, characterized in that: The structure of the strobe head (34) is as follows: the LED light source (35) is electrically connected to the output end of the switching element (27), the control end of the switching element (27) is electrically connected to the controller, and the input end of the switching element (27) is electrically connected to the power supply (29).

5. The high-speed microscopic image acquisition method according to claim 1, characterized in that: An x-axis travel sensor (23) is provided on the stepping stage to verify the relative displacement between the base (26) and the x-axis seat (15) during strobe motion. If an error is detected, the error value is sent and stored as the x-axis correction value for subsequent image stitching. A y-axis travel sensor (21) is provided on the stepping stage to verify the relative displacement between the x-axis seat (15) and the y-axis seat (14) during stroboscopic motion. If an error is detected, the error value is sent and stored as the y-axis correction value for subsequent image stitching.

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