A method for optimizing a scanning path in a high-speed microscopic image acquisition process

Abstract

The application provides a method for optimizing a scanning path in a high-speed microscopic image acquisition process, comprising the following steps: acquiring a panoramic image of a tissue sample on a slide; acquiring an outer contour of the tissue sample; determining a global field range and dividing the global field range into field line frames according to a scanning sequence; overlapping the outer contour of the tissue sample with the global field range; traversing the field line frames according to an "S" shape row-column sequence; marking the field line frames covering the outer contour of the tissue sample as true and marking the field line frames not covering the outer contour of the tissue sample as false; traversing the field line frames marked as true according to the "S" shape row-column sequence to obtain an optimized scanning path. Through the optimization of the scanning path, the number of field line frames that need to be acquired is reduced, the scanning distance is reduced, the amount of subsequent data processing and transmission is reduced, and the scanning and processing efficiency is improved. The processing time of the application is further shortened.

Description

Technical Field

[0001] This invention relates to the field of microscopic image acquisition, and in particular to a method for optimizing the scanning path during high-speed microscopic image acquisition. 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 using this device still takes approximately two hours. The main technical bottleneck lies in the intermittent image acquisition method, requiring a stop 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 employing 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. There are also few dedicated lenses for linear scanners on the market, leading to an exponential increase in image quality and cost. The existing slide scanning path is a rectangular global field of view, while slide samples are typically circular, lacking corresponding sample image information at the corners of the global field of view, which affects efficiency. During the acquisition of pathological tissue images, discontinuous pathological tissue sample images often occur, and there are also areas in other regions without corresponding sample image information. Acquiring images of these areas results in a waste of efficiency. Summary of the Invention

[0003] The technical problem to be solved by the present invention is to provide a method for optimizing the scanning path during high-speed microscopic image acquisition, which can further improve the speed of image acquisition, and can correct the position or imaging quality of the acquired image, thereby 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 method for optimizing the scanning path during high-speed microscopic image acquisition, comprising the following steps:

[0005] S1. Obtain a panoramic image of the tissue sample on the slide;

[0006] S2. Obtain the outer contour of the tissue sample;

[0007] S3. Determine the global field of view and divide the global field of view into various field of view frames according to the scanning order;

[0008] S4. Overlay the outer contour of the tissue sample with the global field of view;

[0009] S5. Traverse the field of view wireframe in an "S" shaped row and column order;

[0010] S6. Mark the field of view that covers the outer contour of the tissue sample as true, and mark the field of view that does not cover the outer contour of the tissue sample as false.

[0011] S7. Traverse the field of view wireframes marked as true in an "S" shape and in row and column order to obtain the optimized scan path.

[0012] A method for optimizing the scanning path during high-speed microscopic image acquisition includes the following steps:

[0013] S01. Obtain a panoramic image of the tissue sample on the slide;

[0014] S02, Obtain the outer contour of each tissue sample;

[0015] S03. Determine the global field of view and divide the global field of view into various field of view frames according to the scanning order;

[0016] S04. Overlay the outer contour of each tissue sample with the global field of view;

[0017] S05. Traverse the field of view wireframe in an "S" shaped row and column order;

[0018] S06. Mark the field of view that covers the outer contour of the tissue sample as true, and mark the field of view that does not cover the outer contour of the tissue sample as false.

[0019] S07. Traverse the field of view wireframes marked as true in an "S" shape and in row and column order;

[0020] S08. Detect the number of tissue samples with their outer contours;

[0021] S09. If the quantity is 1, the scanning path and the corresponding strobe pulse signal are obtained directly.

[0022] If the number is greater than 1, the field of view frame marked as true at both ends of each row is taken as the current scan path, and the corresponding strobe pulse signal is true. The positions of the field of view frame of the current row and the next row are compared, and the farther position is taken as the line break position. The newly added scan path is merged into the current scan path to obtain the optimized scan path and the corresponding strobe pulse signal.

[0023] In a preferred embodiment, the microscopic image acquisition device includes a stepping stage, a microscope tube, a panoramic lens, an industrial camera, a strobe head, and an industrial control computer.

[0024] 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.

[0025] The steps of the image acquisition control method are as follows:

[0026] S11. Calculate the number of step pulse signals based on the range of the field of view frame;

[0027] S12. Within the field of view frame, the sum of the periods of the step pulse signals is taken as the strobe period.

[0028] S13. Continuously send stepping pulse signals to the stepper motor according to the optimized scanning path;

[0029] S14. Based on the strobe pulse signal corresponding to the optimized scanning path, take 1 / 2 strobe cycle and send the strobe pulse signal to the strobe head for the first field of view in the current row;

[0030] Simultaneously send exposure pulse signals to the industrial camera;

[0031] While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view;

[0032] S15. Subsequent fields of view send strobe pulse signals to the strobe head according to the strobe cycle;

[0033] Simultaneously send exposure pulse signals to the industrial camera;

[0034] While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view;

[0035] The first field of view in the next row only calculates the y-axis flicker period, not the x-axis flicker period;

[0036] If there is no corresponding strobe pulse signal in the current field of view, then no strobe pulse signal will be sent to the strobe head;

[0037] S16. Store image data and stitch it together according to coordinates;

[0038] The above steps enable high-speed acquisition of microscopic images.

[0039] In the preferred embodiment, the number of step pulse signals in the x-direction = x-direction field of view frame length / (single step pulse rotation angle / 360) × x-axis screw pitch;

[0040] The number of step pulse signals in the y-direction = y-direction field of view frame width / (single step pulse rotation angle / 360) × y-axis screw pitch.

[0041] In the preferred embodiment, in step S11, the field of view refers to the physical size corresponding to the single-exposure image data on the slide, including the x-axis length and the y-axis length;

[0042] In step S12, the strobe period 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.

[0043] In the preferred embodiment, the width of the strobe pulse signal is greater than the width of the exposure pulse signal, that is, the time for the strobe light to shine is greater than the exposure time.

[0044] 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.

[0045] 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.

[0046] 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.

[0047] 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.

[0048] In the preferred embodiment, step S16 includes the following steps during splicing:

[0049] S61. Combine the preset field of view frames into a complete field of view;

[0050] Convert the scanned field-of-view image to its actual size in pixels;

[0051] S62. Fill the visual field image into the corresponding visual field frame in a centered alignment manner; in the first two visual fields, fill the subsequent visual field frame in the x-direction with an overlay alignment operation, and store the x-direction offset value between the center of the visual field image and the visual field frame as the x-direction filling correction parameter for subsequent visual field images.

[0052] 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.

[0053] S64. When a new line is inserted, two adjacent visual field images are selected. The first visual field image is used as the reference. The image is filled into the visual field frame of the next line in the y direction using an overlay alignment operation. The y-direction offset value between the center of the visual field image and the visual field frame is stored as the y-direction filling correction parameter for the subsequent visual field image. If there is no visual field image in the current visual field frame, the correction is performed on the visual field frame where there is a visual field image.

[0054] 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.

[0055] The above steps enable rapid stitching of visual field images.

[0056] In a preferred embodiment, a light compensation sensor is also provided. In steps S14 and S15, the illuminance value of the stroboscopic flash collected by the light compensation sensor is synchronously acquired according to the exposure pulse signal. In step S16, 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.

[0057] The present invention provides a method for optimizing the scanning path during high-speed microscopic image acquisition. Compared with the prior art, the method employing the above-mentioned technical solution has the following advantages:

[0058] 1. This invention, through optimization of the scanning path, reduces the number of field-of-view frames to be acquired, shortens the scanning distance, reduces subsequent data processing and transmission, and improves scanning and processing efficiency. Compared with the inventors' continuous scanning and stitching scheme, the processing time of this invention is further reduced by 13 seconds.

[0059] Furthermore, for non-continuous pathological tissue images, it can further reduce the required field of view outlines, reduce scanning distance, reduce subsequent data processing and transmission, and improve scanning and processing efficiency.

[0060] 2. Compared with existing line scanning schemes, 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, this significantly improves image acquisition efficiency, reducing acquisition time from over 20 minutes to 58-120 seconds (depending on the number of fields of view). Figure 11 As shown, the image with 1392 fields of view, or 29×48 fields of view, was scanned and stitched in just 58 seconds, reaching the international leading level.

[0061] 2. By adopting an optimized continuous walking + on-demand stroboscopic scheme, the microscope tube can use a smaller numerical aperture, resulting in a deeper field of view for the acquired images, improved diffraction-limited image accuracy, and less image edge distortion and dispersion.

[0062] 4. By using a strobe method, the heat dissipation pressure of the lighting source is reduced due to the shorter working time, and a higher power LED light source can be used. The preferred working illuminance of the LED light source of this invention is 100,000 lx.

[0063] 5. An open-loop control scheme combined with post-processing error correction was adopted, which further improved the acquisition speed while ensuring splicing accuracy.

[0064] 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.

[0065] 7. The method of the present invention significantly reduces equipment costs and improves efficiency through precise control and simplified algorithms, making it more conducive to the promotion of microscopic scanners in primary medical institutions, reducing the cost of use for each user, and intercepting malignant tumors such as cervical cancer in the initial stage, thus greatly improving the cure rate of tumors.

[0066] 8. The method of the present invention is particularly suitable for scanning pathological tissue samples with discontinuous images, and can further improve the efficiency of scanning acquisition. Attached Figure Description

[0067] The present invention will be further described below with reference to the accompanying drawings and embodiments:

[0068] Figure 1 This is a comparative diagram showing the scanning path before and after optimization according to the present invention.

[0069] Figure 2 This is a flowchart illustrating the scanning path optimization of the present invention.

[0070] Figure 3 This is a comparative diagram showing the preferred scanning path before and after optimization according to the present invention.

[0071] Figure 4 The flowchart illustrates the preferred scanning path optimization method of this invention.

[0072] Figure 5 This is a flowchart of the image acquisition process in this invention.

[0073] Figure 6 This is a schematic diagram of the synchronization control pulse signal of the present invention.

[0074] Figure 7 This is a schematic diagram of the overall structure of the present invention.

[0075] Figure 8 This is a perspective view of the stage support of the present invention.

[0076] Figure 9 This is a schematic diagram of the structure of the strobe head of the present invention.

[0077] Figure 10 This is a schematic diagram of the glass slide structure of the present invention.

[0078] Figure 11 This is a schematic diagram of the scanning result interface of the method of the present invention.

[0079] Figure 12 This is a schematic diagram of the scanning result interface of the preferred method of the present invention.

[0080] 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 cycle; 36. Field of view 100; Scanning path 200; New scan path 201; Field of view frame 300; Tissue sample outer contour 400. Detailed Implementation

[0081] Example 1:

[0082] The following are examples of the device used in this invention; however, the method of this invention is not limited to use on the device described above. Figure 7 , 8 In this invention, a high-speed microscopic image acquisition device includes a stepping stage, a microscope tube 10, a panoramic lens 12, an industrial camera 33, a strobe head 34, a memory 4, a controller 3, and an industrial computer 2.

[0083] A panoramic lens 12 is mounted on a microscope tube 10, which is connected to an industrial camera 33. 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 emits 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.

[0084] 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.

[0085] 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;

[0086] 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.

[0087] 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.

[0088] 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 11As 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 58 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.

[0089] 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.

[0090] 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.

[0091] Example 2:

[0092] like Figure 1 , 2 A method for optimizing the scanning path during high-speed microscopic image acquisition includes the following steps:

[0093] S1. Obtain a panoramic image of the tissue sample 32 on the slide 31; preferably, the panoramic image is acquired through a panoramic lens 12, which is also used to acquire an coded image 30 on the slide 31, such as... Figure 10 As shown in the image.

[0094] S2. Obtain the outer contour of tissue sample 32; preferably, by binarizing the panoramic image of tissue sample 32, the outer contour of tissue sample 32 can be obtained relatively easily based on edge contrast. Typically, the outer contour of tissue sample 32 is fitted to a circle, for example... Figure 1 , 3 The outer contour of the tissue sample shown in Figure 11 is 400.

[0095] S3. Determine the global field of view and divide it into individual field of view frames 300 according to the scanning order; the global field of view is the smallest rectangle including the tissue sample 32. Based on the single field of view 100 of the panoramic lens 12, the global field of view is divided into individual field of view frames 300. Typically, the length and width of the field of view 100 are greater than or equal to the length and width of the field of view frame 300.

[0096] S4. Overlay the outer contour of the tissue sample (400°) with the global field of view, such as... Figure 1 As shown in the image.

[0097] S5. Traverse the view frame in an "S" shaped row and column order; the "S" shaped row and column order means traversing the first row from left to right, then moving down to the next row, traversing the second row from right to left, and so on.

[0098] S6. Mark the field of view 300 that covers the outer contour 400 of the tissue sample as true, and mark the field of view 300 that does not cover the outer contour 400 of the tissue sample as false.

[0099] S7. Traverse the field-of-view wireframes marked as true (300) in an "S" shape and row / column order to obtain the optimized scan path. It should be noted that... Figure 1 The numbers shown are for illustrative purposes only, mainly for ease of observation. The actual number of 300 frames in the field of view is much greater than that. Figure 1 The number of field-of-view wireframes 300 shown in the figure. (By...) Figure 1 As shown, the scan path 200 in the lower figure is approximately 22% shorter than the scan path in the upper figure, resulting in a significant reduction in the amount of image data transmitted, stored, and computed. For large-scale image acquisition, such as millions of images, this increase in efficiency is very significant.

[0100] Example 3:

[0101] like Figure 3 , 4 A method for optimizing the scanning path during high-speed microscopic image acquisition includes the following steps:

[0102] S01. Obtain a panoramic image of tissue sample 32 on slide 31;

[0103] S02. Obtain the outer contour of each tissue sample 32; for some pathological tissues, many are fragmented images. Therefore, in this case, each tissue sample 32 has an independent outer contour. Mark and count the number of outer contours 400 of each tissue sample.

[0104] S03. Determine the global field of view and divide the global field of view into various field of view frames 300 according to the scanning order;

[0105] S04. Overlap the outer contour 400 of each tissue sample with the global field of view;

[0106] S05. Traverse the field of view wireframe in an "S" shaped row and column order;

[0107] S06. Mark the field of view 300 that covers the outer contour 400 of the tissue sample as true, and mark the field of view 300 that does not cover the outer contour 400 of the tissue sample as false.

[0108] S07. Traverse the field of view wireframes marked as true in an "S" shape and in row and column order;

[0109] S08. Detect the number of tissue samples with an outer contour of 400.

[0110] S09. If the quantity is 1, the scanning path and the corresponding strobe pulse signal are obtained directly.

[0111] If the number is greater than 1, the true field of view frame 300 at both ends of each row is taken as the current scan path. If there is a false field of view frame 300 between the two field of view frames 300 at the end, it is added to the current scan path as a new scan path 201.

[0112] The strobe pulse signal corresponding to the field of view frame 300 marked as true is true, and the strobe pulse signal corresponding to the corresponding field of view frame 300 marked as false is false. That is, when the microscope tube 10 passes through the field of view frame 300, the industrial control computer 2 does not send a strobe pulse signal to the strobe head 34, nor does it send an exposure pulse signal to the industrial camera 33. The positions of the field of view frames 300 in the current row and the next row are compared, and the position that is farther away, such as the position of the field of view frame 300 that is closer to the outside, is taken as the line break position. Figure 3 As shown, the scan path traversing the false field of view frame 300 is treated as a new scan path 201. This new scan path 201 is then merged into the current scan path, resulting in an optimized scan path and the corresponding strobe pulse signal. For ease of description, the corresponding strobe pulse signal naturally also includes the corresponding exposure pulse signal. This method is particularly suitable for scanning pathological tissue samples with discontinuous images, further improving the efficiency of scan acquisition.

[0113] Example 4:

[0114] In Example 2 or Example 3, the steps of the image acquisition control method are as follows:

[0115] S11. Calculate the number of step pulse signals based on the 300-degree field of view frame.

[0116] In the preferred embodiment, the number of step pulse signals in the x-direction length = x-direction field of view frame length 300 / (single step pulse rotation angle / 360) × x-axis screw pitch;

[0117] The number of step pulse signals in the y-direction length = y-direction field of view frame width 300 / (single step pulse rotation angle / 360) × y-axis screw pitch.

[0118] In the preferred embodiment, 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;

[0119] S12. Within the field of view frame of 300, the sum of the periods of the step pulse signals is taken as the strobe period 36.

[0120] The strobe cycle 36 includes an x-axis strobe cycle and a y-axis strobe cycle. The x-axis strobe cycle is the sum of the periods of the step pulse signals along the x-axis length, and the y-axis strobe cycle is the sum of the periods of the step pulse signals along the y-axis length. That is, when the computer 2 reaches the center position of each field of view frame 300, it sends a strobe pulse signal to the strobe head 34. In other words, the strobe cycle 36 is the sum of the periods of the step pulse signals within the travel range of a field of view frame 300. For example, if the travel time for a field of view 100 is 19ms, then the corresponding strobe cycle 36 is 19ms.

[0121] In the preferred embodiment, the width of the strobe pulse signal is greater than the width of the exposure pulse signal, that is, the time for the strobe head 34 to emit light is greater than the exposure time;

[0122] 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, preferably 19 ms, 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.

[0123] In a preferred embodiment, the structure of the strobe head 34 is such that 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.

[0124] S13. Stepping pulse signals are continuously sent to the stepper motors according to the optimized scanning path; 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.

[0125] S14. Based on the strobe pulse signal corresponding to the optimized scanning path, take 1 / 2 strobe cycle 36 for the first field of view 100 in the current row and send a strobe pulse signal to the strobe head; the strobe head's illumination duration is 30~60 microseconds;

[0126] 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.

[0127] While the strobe head flashes, the industrial camera exposes and acquires 100 image data of the current field of view;

[0128] In a preferred embodiment, a light compensation sensor 11 is also provided, which synchronously acquires the illuminance value of the stroboscopic flash based on the exposure pulse signal.

[0129] S15, The subsequent field of view 100 sends a strobe pulse signal to the strobe head according to a strobe cycle of 36;

[0130] Simultaneously send exposure pulse signals to the industrial camera;

[0131] While the strobe head flashes, the industrial camera exposes and acquires 100 image data of the current field of view;

[0132] The first field of view in the next row is 100. Only the y-direction flicker period is calculated, and the x-direction flicker period is not calculated.

[0133] If the current field of view is 100 and there is no corresponding strobe pulse signal, then no strobe pulse signal will be sent to the strobe head;

[0134] While the strobe head flashes, the industrial camera exposes and acquires 100 image data of the current field of view;

[0135] 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.

[0136] S16. Store image data and stitch it together according to coordinates;

[0137] The above steps enable high-speed acquisition of microscopic images.

[0138] In a preferred embodiment, 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. If an error is detected, the error value is sent and stored as the x-axis correction value for subsequent image stitching.

[0139] The stepping stage is equipped with a y-axis travel sensor 21, which is used to verify the relative displacement between the x-axis seat 15 and the y-axis seat 14. If an error is detected, the error value is sent and stored as the y-axis correction value for subsequent image stitching.

[0140] In the preferred embodiment, step S16 includes the following steps during splicing:

[0141] S61. Stitch together the preset field of view frame 300 to form a complete field of view;

[0142] Convert the scanned field-of-view image to its actual size in pixels;

[0143] S62. Fill the field of view image into the corresponding field of view frame 300 in a centered alignment manner; in the first two fields of view 100, fill the subsequent field of view frame 300 in the x direction with an overlay alignment operation, and store the x-direction offset value between the center of the field of view image and the field of view frame 300 as the x-direction filling correction parameter for the subsequent field of view image.

[0144] Center alignment refers to 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; the excess portion after filling is clipped by the field of view frame 300. Subsequent field of view images are filled into the subsequent field of view frame 300 using a superposition alignment operation in the x-direction, based on the preceding field of view image. The superposition alignment operation calculates pixels with identical or similar edges in 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 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 subsequent field of view images.

[0145] 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 back field of view image.

[0146] S64. When a new line breaks, select two adjacent visual field images. Using the preceding visual field image as a reference, fill the following visual field frame 300 in the y-direction with an overlay alignment operation. Store the y-direction offset value between the center of the visual field image and the visual field frame 300 as the y-direction filling correction parameter for the subsequent visual field image. If the current visual field frame 300 has no visual field image, then the correction is performed on the visual field frame 300 with a visual field image.

[0147] 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.

[0148] 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 the stroboscopic lights in 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.

[0149] The above steps enable rapid stitching of visual field images.

[0150] 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 method for optimizing the scanning path during high-speed microscopic image acquisition, characterized in that: Includes the following steps: S1. Obtain a panoramic image of the tissue sample (32) on the slide (31); S2. Obtain the outer contour of the tissue sample (32); S3. Determine the global field of view and divide the global field of view into various field of view wireframes (300) according to the scanning order. S4. Overlay the outer contour (400) of the tissue sample with the global field of view; S5. Traverse the field of view wireframe in an "S" shaped row and column order; S6. Mark the field of view (300) that covers the outer contour (400) of the tissue sample as true, and mark the field of view (300) that does not cover the outer contour (400) of the tissue sample as false; S7. Traverse the field of view (300) marked as true in an "S" shape and in row and column order to obtain the optimized scan path.

2. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 1, characterized in that: The microscopic image acquisition device includes a stepping stage, a microscope tube (10), a panoramic lens (12), an industrial camera (33), a strobe head (34), and an industrial control computer (2). 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). The steps of the image acquisition control method are as follows: S11. Calculate the number of step pulse signals based on the range of the field of view frame (300); S12. Within the field of view frame (300), the sum of the periods of the step pulse signals is taken as the strobe period (36). S13. Continuously send stepping pulse signals to the stepper motor according to the optimized scanning path; S14. Based on the strobe pulse signal corresponding to the optimized scanning path, take 1 / 2 strobe cycle (36) of the first field of view (100) of the current row and send the strobe pulse signal to the strobe head; Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view (100); S15, The subsequent field of view (100) sends a strobe pulse signal to the strobe head according to the strobe cycle (36); Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view (100); The first field of view (100) in the next row only calculates the y-direction flicker period, not the x-direction flicker period; If there is no corresponding strobe pulse signal in the current field of view (100), then no strobe pulse signal will be sent to the strobe head; S16. Store image data and stitch it together according to coordinates; The above steps enable high-speed acquisition of microscopic images.

3. A method for optimizing the scanning path during high-speed microscopic image acquisition, characterized by: Includes the following steps: S01. Obtain a panoramic image of the tissue sample (32) on the slide (31); S02, Obtain the outer contour of each tissue sample (32); S03. Determine the global field of view and divide the global field of view into various field of view wireframes (300) according to the scanning order. S04. Overlay the outer contour (400) of each tissue sample with the global field of view; S05. Traverse the field of view wireframe in an "S" shaped row and column order; S06. Mark the field of view (300) that covers the outer contour (400) of the tissue sample as true, and mark the field of view (300) that does not cover the outer contour (400) of the tissue sample as false; S07. Traverse the field of view (300) marked as true in an "S" shape and in row and column order. S08, Detect the number of tissue samples with an outer contour (400); S09. If the quantity is 1, the scanning path and the corresponding strobe pulse signal are obtained directly. If the number is greater than 1, the field of view (300) marked as true at both ends of each row is taken as the current scanning path, and the corresponding strobe pulse signal is true. The positions of the field of view (300) of the current row and the next row are compared, and the more distant position is taken as the line break position. The newly added scanning path (201) is merged into the current scanning path to obtain the optimized scanning path and the corresponding strobe pulse signal.

4. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 3, characterized in that: The microscopic image acquisition device includes a stepping stage, a microscope tube (10), a panoramic lens (12), an industrial camera (33), a strobe head (34), and an industrial control computer (2). 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). The steps of the image acquisition control method are as follows: S11. Calculate the number of step pulse signals based on the range of the field of view frame (300); S12. Within the field of view frame (300), the sum of the periods of the step pulse signals is taken as the strobe period (36). S13. Continuously send stepping pulse signals to the stepper motor according to the optimized scanning path; S14. Based on the strobe pulse signal corresponding to the optimized scanning path, take 1 / 2 strobe cycle (36) of the first field of view (100) of the current row and send the strobe pulse signal to the strobe head; Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view (100); S15, The subsequent field of view (100) sends a strobe pulse signal to the strobe head according to the strobe cycle (36); Simultaneously send exposure pulse signals to the industrial camera; While the stroboscopic head flashes, the industrial camera exposes and acquires image data of the current field of view (100); The first field of view (100) in the next row only calculates the y-direction flicker period, not the x-direction flicker period; If there is no corresponding strobe pulse signal in the current field of view (100), then no strobe pulse signal will be sent to the strobe head; S16. Store image data and stitch it together according to coordinates; The above steps enable high-speed acquisition of microscopic images.

5. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 4, characterized in that: The number of step pulse signals in the x-direction = x-direction field of view frame (300) 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 frame (300) width / (single step pulse rotation angle / 360) × y-axis screw pitch.

6. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 4, characterized in that: In step S11, 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; In step S12, 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.

7. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 4, 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 strobe head (34) to emit light is greater than the exposure time; 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.

8. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 4, 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).

9. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 4, 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.

10. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 9, characterized in that: Step S16 includes the following steps during splicing: S61. Piece together the preset field of view frame (300) to form a complete field of view; Convert the scanned field-of-view image to its actual size in pixels; S62. Fill the field of view image into the corresponding field of view frame (300) in a center-aligned manner; in the first two fields of view (100), fill the field of view frame (300) in the x-direction with an overlay alignment operation, and store the x-direction offset value between the center of the field of view image and the field of view frame (300) 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, two adjacent visual field images are selected. The first visual field image is used as the reference. The image is filled in the y-direction using an overlay alignment operation to the visual field frame (300) in the next line. The y-direction offset value between the center of the visual field image and the visual field frame (300) is stored as the y-direction filling correction parameter for the subsequent visual field image. If there is no visual field image in the current visual field frame (300), the correction is performed in the visual field frame (300) where there is a visual field 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 rapid stitching of visual field images.

11. The method for optimizing the scanning path during high-speed microscopic image acquisition according to claim 9, characterized in that: A light compensation sensor (11) is also provided. In steps S14 and S15, 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 S16, 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.

Images