Method and system for searching and positioning of chromosome karyotypes

Abstract

The application provides a searching and positioning method and system for chromosome karyotype, comprising: scanning a chromosome sample to collect a microscopic image; detecting and recording a coordinate position of the chromosome karyotype according to the microscopic image; establishing a spatial coordinate system to obtain an estimated coordinate position of the chromosome karyotype under a target objective lens; moving a target chromosome karyotype to the estimated coordinate position; performing grid imaging on the target chromosome karyotype to match the target chromosome karyotype at different magnifications, calculating an actual coordinate position of the target chromosome karyotype based on a picture matching result, obtaining a deviation between the actual coordinate position and the estimated coordinate position, moving the target chromosome karyotype to a center field of view, collecting and saving an image of the target chromosome karyotype. The target karyotype is placed in the center field of view under the high-magnification objective lens through position deviation correction, and clear imaging is performed, so that the positioning efficiency and positioning accuracy of the chromosome karyotype are improved by more than 50%.

Description

Technical Field

[0001] This invention belongs to the field of image processing and pattern recognition technology, specifically relating to a method and system for rapid karyotype retrieval and localization based on multi-scale feature fusion. More specifically, this invention provides a solution for achieving efficient karyotype search and accurate localization through feature extraction and multi-scale analysis. Background Technology

[0002] In biomedical research and clinical diagnosis, chromosome karyotype analysis is a crucial technique, playing an indispensable role in elucidating the pathogenesis of genetic diseases, prenatal diagnosis, and cancer research. Accurate identification and precise localization of the studyable metaphase chromosome karyotype are fundamental to effective analysis.

[0003] Currently, the combined high- and low-power microscopy method for precise localization is one of the commonly used techniques in chromosome karyotype analysis. Specifically, this method first uses push-broom microscopy under a low-power objective to perform a large-scale scan and coarse localization of the chromosome karyotype on the sample slice; then, it switches to a high-power objective for high-resolution microscopy to further refine the localization and obtain clear microscopic images. However, due to factors such as transmission system errors, insufficient rigidity of the guide mechanism, and limitations of the drive control system, the precise localization of the same chromosome karyotype across scales can be inaccurate, leading to deviations in cross-scale target localization and consequently affecting subsequent high-power microscopic imaging. In other words, the low-power objective has a larger field of view, while the high-power objective has a smaller field of view. If the cross-scale repetitive localization deviation is large when switching to the high-power objective, the same karyotype located under the low-power objective may not be observable under the high-power objective field of view. Furthermore, there is no consistency verification of karyotypes under different fields of view, resulting in a lack of sufficient localization accuracy.

[0004] It is evident that the efficiency and practicality of existing technologies are limited, making it difficult to meet the demands for high-precision and high-efficiency chromosome karyotype analysis.

[0005] However, while existing microscopic imaging devices for chromosome karyotype analysis have improved target localization accuracy to some extent, they still have several shortcomings. Patent CN119165640A discloses a microscope objective identification device and method, solving the technical challenge of automatic identification and switching of microscope objectives; Patent CN115079393A discloses a microscopic imaging device and method for chromosome karyotype analysis based on a 10x objective, which improves the accuracy of subsequent 100x objective imaging by optimizing coordinate detection under the 10x objective. However, none of these technical solutions solve the problem of target karyotype loss caused by differences in the field of view during objective switching, nor can they ensure the consistency of the target karyotype before and after objective switching.

[0006] Patent document CN119168856A discloses an intelligent microscopic image stitching system. This system includes a microscopic image acquisition unit and an image stitching unit. The image stitching unit first stitches and repairs the microscopic images of the entire field of view of the slide acquired under the highest magnification objective lens to obtain a seamless, complete full-field image under the highest magnification objective lens. Then, based on the seamless, complete full-field image under the highest magnification objective lens, it stitches and repairs the microscopic images under other magnification objectives to obtain seamless, complete full-field images under the remaining objectives. However, this system also fails to solve the problem of target karyotype loss caused by differences in the field of view when switching objectives, and it cannot ensure the consistency of the target karyotype before and after objective lens switching. This problem urgently needs to be solved. Summary of the Invention

[0007] In view of the deficiencies in the prior art, the purpose of this invention is to provide a method and system for searching and locating chromosome karyotypes.

[0008] A method for searching and locating chromosome karyotypes according to the present invention includes:

[0009] Step S1: Scan the chromosome sample and acquire microscopic images under a low-power objective lens;

[0010] Step S2: Based on the microscopic image, detect and record the coordinate position of the chromosome karyotype;

[0011] Step S3: Establish a spatial coordinate system, and then obtain the estimated coordinate position of the chromosome karyotype under a high-power objective lens;

[0012] Step S4: Under a high-power objective lens, move the target chromosome karyotype to the estimated coordinate position;

[0013] Step S5: Perform image matching of the same target karyotype at different magnifications and determine whether the matching is successful. If the result is yes, obtain the image matching result and proceed to step S6; if the result is no, perform expanded grid imaging with the current field of view center, stitch the grid images, and repeat step S5 until the grid image of the target chromosome karyotype is successfully matched.

[0014] Step S6: Based on the image matching results, calculate the actual coordinate position of the target chromosome karyotype, and then determine the deviation between the actual coordinate position and the estimated coordinate position. Move the target chromosome karyotype to the center of the field of view; acquire and save the image of the target chromosome karyotype.

[0015] Preferably, in step S2, the coordinates of the metaphase chromosome karyotype are detected and recorded using the Faster-RCNN object detection model.

[0016] Preferably, in step S3, the estimated coordinate position of the chromosome karyotype is expressed mathematically as follows:

[0017]

[0018] Among them, A x Estimate the physical coordinates of the target point along the X-axis under low magnification; A y Estimate the physical coordinates of the target point along the Y-axis under low magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; S x The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; S y The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; V x V represents the physical width of the sample slice image. y is the physical height of the sample slice image; W is the horizontal pixel count of the sample slice image; H is the vertical pixel count of the sample slice image.

[0019] Preferably, in step S5, the SIFT algorithm is used to match the target chromosome karyotype at different magnifications;

[0020] The process of stitching the grid images includes:

[0021] Step A1: Calculate the scan step distance and generate scan grid points; the scan grid points are initialized at (x0, y0) to generate a 3×3 scan matrix;

[0022] Step A2: Stitch and expand the grid imaging;

[0023] In step A1, the mathematical expression for the scanning step distance is:

[0024]

[0025] Where Δx is the step size in the X direction, Δy is the step size in the Y direction; w is the width of the rectangular ROI region; h is the height of the rectangular ROI region; α is the overlap coefficient; the symbol · represents the product;

[0026] The mathematical expression for the coordinates of the scanned grid points is:

[0027]

[0028] Where, x ij Let y be the x-coordinate of the scanned grid point. ij The ordinate of the scan grid point is (x0, y0); (x0, y0) is the initial point, i.e., the center; i and j are the grid index positions used to identify the specific positions, i, j∈{0,1,2};

[0029] In step A2, the total coverage area size of the expanded grid imaging is mathematically expressed as:

[0030]

[0031] α is the overlap coefficient between adjacent fields of view: α∈(0,1); Δx is the displacement of the stage in the X direction; Δy is the displacement of the stage in the Y direction; W″ is the total width of the stitched image; H″ is the total height of the stitched image.

[0032] Preferably, in step S6, the mathematical expression for the actual coordinate position is:

[0033]

[0034] Among them, B x B represents the actual physical coordinates of the target point along the X-axis under high magnification; y A represents the actual physical coordinates of the target point along the Y-axis under high magnification. x To estimate the physical coordinates along the X-axis under high magnification; A y To estimate the physical coordinates along the Y-axis under high magnification; S x ′ represents the X-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; S y ′ represents the Y-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; V x ′ represents the physical width of the stitched images; V y ′ represents the physical height of the stitched image; W′ represents the horizontal pixel count of the stitched image; H′ represents the vertical pixel count of the stitched image.

[0035] The mathematical expression for the deviation between the actual coordinate position and the estimated coordinate position is:

[0036] Δx'=B x -A x

[0037] Δy'=B y -A y

[0038] Where Δx' is the deviation in the X-axis direction and Δy' is the deviation in the Y-axis direction.

[0039] A system for searching and locating chromosome karyotypes according to the present invention includes:

[0040] Module M1: Scans chromosome samples and acquires microscopic images under a low-power objective lens;

[0041] Module M2: Based on the microscopic image, detect and record the coordinate position of the chromosome karyotype;

[0042] Module M3: Establishes a spatial coordinate system to obtain the estimated coordinate position of the chromosome karyotype under a high-power objective lens;

[0043] Module M4: Under high magnification, move the target chromosome karyotype to the estimated coordinate position;

[0044] Module M5: Performs image matching of the same target karyotype at different magnifications and determines whether the matching is successful. If the result is yes, the image matching result is obtained, and module M6 is triggered; if the result is no, extended grid imaging is performed with the current field of view center, the grid image is stitched together, and module M5 is triggered again until the grid image of the target chromosome karyotype is successfully matched.

[0045] Module M6: Based on the image matching results, calculate the actual coordinate position of the target chromosome karyotype, then determine the deviation between the actual coordinate position and the estimated coordinate position, move the target chromosome karyotype to the center of the field of view; acquire and save the image of the target chromosome karyotype.

[0046] Preferably, in module M2, the coordinate positions of the metaphase chromosome karyotype are detected and recorded using the Faster-RCNN object detection model.

[0047] Preferably, in module M3, the estimated coordinate position of the chromosome karyotype is expressed mathematically as follows:

[0048]

[0049] Among them, A x Estimate the physical coordinates of the target point along the X-axis under low magnification; A y Estimate the physical coordinates of the target point along the Y-axis under low magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; S x The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; S y The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; V x V represents the physical width of the sample slice image. y is the physical height of the sample slice image; W is the horizontal pixel count of the sample slice image; H is the vertical pixel count of the sample slice image.

[0050] Preferably, in module M5, the SIFT algorithm is used to match the image with the target chromosome karyotype at different magnifications;

[0051] The stitched grid image includes:

[0052] Module A1: Calculates the scan step distance and generates scan grid points; the scan grid points are initialized at (x0, y0) to generate a 3×3 scan matrix;

[0053] Module A2: Stitching and expanding grid imaging;

[0054] In module A1, the mathematical expression for the scanning step distance is:

[0055]

[0056] Where Δx is the step size in the X direction, Δy is the step size in the Y direction; w is the width of the rectangular ROI region; h is the height of the rectangular ROI region; α is the overlap coefficient; the symbol · represents the product;

[0057] The mathematical expression for the coordinates of the scanned grid points is:

[0058]

[0059] Where, x ij Let y be the x-coordinate of the scanned grid point. ij The ordinate of the scan grid point is (x0, y0); (x0, y0) is the initial point, i.e., the center; i and j are the grid index positions used to identify the specific positions, i, j∈{0,1,2};

[0060] In module A2, the total coverage area size of the extended grid imaging is mathematically expressed as:

[0061]

[0062] α is the overlap coefficient between adjacent fields of view: α∈(0,1); Δx is the displacement of the stage in the X direction; Δy is the displacement of the stage in the Y direction; W″ is the total width of the stitched image; H″ is the total height of the stitched image.

[0063] Preferably, in module M6, the mathematical expression for the actual coordinate position is:

[0064]

[0065] Among them, B x B represents the actual physical coordinates of the target point along the X-axis under high magnification; y A represents the actual physical coordinates of the target point along the Y-axis under high magnification. x To estimate the physical coordinates along the X-axis under high magnification; A y To estimate the physical coordinates along the Y-axis under high magnification; S x ′ represents the X-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; S y′ represents the Y-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; V x ′ represents the physical width of the stitched images; V y ′ represents the physical height of the stitched image; W′ represents the horizontal pixel count of the stitched image; H′ represents the vertical pixel count of the stitched image.

[0066] The mathematical expression for the deviation between the actual coordinate position and the estimated coordinate position is:

[0067] Δx'=B x -A x

[0068] Δy'=B y -A y

[0069] Where Δx' is the deviation in the X-axis direction and Δy' is the deviation in the Y-axis direction.

[0070] Compared with the prior art, the present invention has the following beneficial effects:

[0071] 1. This invention uses chromosome extended grid imaging to rapidly locate target karyotypes under high magnification objectives. Specifically, it determines the consistency of the target karyotype under different magnifications by comparing the feature similarity of metaphase chromosome karyotypes captured at high and low magnifications.

[0072] 2. This invention places the target karyotype in the central field of view under a high-power objective lens through positional deviation correction, enabling clear imaging and improving the localization efficiency and accuracy of chromosome karyotypes by more than 50%.

[0073] 3. This invention uses a cross-scale image registration algorithm to match the target kernel image and obtain the actual coordinate position of the target kernel, thus solving the problem of losing the target kernel location due to the difference in the field of view of imaging of different magnification objectives. Attached Figure Description

[0074] Other features, objects, and advantages of the present invention will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0075] Figure 1 This is a schematic diagram of the method flow provided by the present invention;

[0076] Figure 2 A flowchart of cross-scale spatial coordinate system transformation provided by the present invention;

[0077] Figure 3 A flowchart of extended grid imaging under high-magnification objectives provided by the present invention;

[0078] Figure 4A schematic diagram of the structure of the chromosome karyotype microscopic scanning imaging device provided by the present invention;

[0079] Figure 5 This is a schematic diagram of the positional relationship of chromosome karyotype images under a low-power objective lens provided by the present invention.

[0080] Figure 6 A schematic diagram illustrating the calculation of coordinate positions for chromosome karyotype estimation under a high-power objective lens, provided by this invention;

[0081] Figure 7 This is a schematic diagram of an extended grid image under a high-magnification objective lens provided by the present invention.

[0082] Figure label:

[0083] 11 cameras, 21 low-magnification micrographs

[0084] 12 Optical Path Correction System 22 ROI Metaphase Chromosome Karyotype

[0085] 13 Electro-optical microscope converter 31 Current field of view center

[0086] 14 low-power objective lens, 32 target nuclei

[0087] 15mm high-magnification objective lens, 33-grid extended mosaic

[0088] 16 Electric XY Platform

[0089] 17 is a slice of chromosome sample.

[0090] 18 is the instrument base Detailed Implementation

[0091] The present invention will now be described in detail with reference to specific embodiments. These embodiments will help those skilled in the art to further understand the present invention, but do not limit the invention in any way. It should be noted that those skilled in the art can make several changes and improvements without departing from the concept of the present invention. These all fall within the protection scope of the present invention.

[0092] This invention first obtains whole-slice images of chromosome samples under low magnification, searches for metaphase chromosome karyotypes based on a target detection model, and records the coordinate positions of all karyotypes. Then, based on a spatial coordinate system transformation algorithm under different magnification objectives, the estimated coordinate position of a target karyotype under high magnification is obtained. Next, under high magnification, a motorized XY stage is driven to place the target karyotype in the high magnification objective field of view according to the estimated coordinate position, and extended grid imaging is performed. After stitching the grid images, a cross-scale image registration algorithm is used to match the target karyotype image to obtain the actual coordinate position of the target karyotype. Finally, through positional deviation correction, the target karyotype is placed in the central field of view under high magnification for clear imaging.

[0093] Specifically, according to the present invention, a method for searching and locating chromosome karyotypes includes:

[0094] Low-power chromosome sample whole section scanning: Under a low-power objective lens, the sample is moved to the initial position and automatically focused to obtain a clear image. The whole section is automatically scanned according to a predetermined path to obtain high-quality low-power microscopic images.

[0095] Metaphase chromosome karyotype target detection and karyotype coordinate position recording: The target detection model is used to search for metaphase chromosome karyotypes on low-magnification micrographs and the coordinate positions of all karyotypes are recorded;

[0096] Spatial coordinate system transformation and calculation of kernel estimation coordinate position: A spatial coordinate system containing the physical spatial relationship between the instrument base, stage, sample slice and image is established under different magnification objectives. The estimated coordinate position of the kernel under the high magnification objective is obtained according to the spatial coordinate system transformation algorithm under different magnification objectives.

[0097] Objective lens switching and sample slice displacement: Return the motorized Z-axis to zero, switch from low-power objective lens to high-power objective lens, and drive the motorized XY stage to place a target karyotype on the sample slice under the field of view of the high-power objective lens according to the estimated coordinate position;

[0098] Extended grid imaging and cross-scale image matching of target keratin: Autofocus and microscopic imaging are performed under high magnification objectives. A cross-scale image registration algorithm is used to match images of the same target keratin at different magnifications. If the target keratin image matching is not achieved, extended grid imaging is performed with the current field of view center. The grid images are stitched together and then matched until the target keratin image matching is successful.

[0099] Calculation of actual coordinate position of the target kernel and correction of coordinate deviation: The actual coordinate position of the target kernel under the high magnification objective is calculated based on the image matching results and coordinate system transformation algorithm. The actual coordinate position of the target kernel under the high magnification objective is compared with the estimated coordinate position. The coordinate position deviation in the X and Y directions is calculated. Then, the electric XY stage is driven to move the target kernel to the center of the field of view of the high magnification objective.

[0100] High-magnification metaphase chromosome karyotype imaging: Automatic focusing and microscopic imaging of the target karyotype image at the center of the field of view, and saving the image in common image formats such as jpg and png.

[0101] Specifically, the electric biological microscope is a chromosome karyotype microscopic scanning imaging device, including a motor, an electric XY stage, an electric objective lens converter, a 10x objective lens, a 100x objective lens, an electric Z-axis, a camera, an LED light source, a multi-axis motion driver, and other main components.

[0102] Specifically, the whole-slice scanning of the low-power chromosome sample includes the following steps:

[0103] Place the sample slice containing the chromosome karyotype on the motorized XY stage and complete the initial setup of the motorized biological microscope.

[0104] Under a 10x objective lens, the motorized XY stage is driven to move the chromosome sample slice to the initial scanning position, and the image is automatically focused so that the camera can obtain a clear image of a single field of view.

[0105] Using a coordinated LED light source for exposure, ensuring uniform and shadow-free image exposure, a full-slice automatic scan is performed according to a planned path to acquire high-quality 10x magnification microscopic images. Specifically, numerical limitations have been specified for the specific embodiments: low magnification is 10x, high magnification is 100x, and high magnification refers to image quality, which is independent of the magnification.

[0106] Specifically, the metaphase chromosome karyotype target detection and karyotype coordinate position recording includes the following steps:

[0107] The acquired microscopic images are analyzed based on a target detection model to identify metaphase chromosome karyotypes that are elongated, highly dispersed, and have low overlap. The model automatically searches for all karyotypes in the image, accurately locates the position of each karyotype, and records the coordinates and ROI regions of all detected karyotypes.

[0108] Specifically, the spatial coordinate system transformation and kernel estimation coordinate position calculation include the following steps:

[0109] A spatial coordinate system was established to establish the physical spatial relationship between the instrument base, stage, sample slide, and image under two different magnification objectives. This system covers the spatial positioning relationship in four dimensions, including the relative positional relationship between the stage and the instrument base, the relative positional relationship between the chromosome sample slide and the stage, the relative positional relationship between the image and the chromosome sample slide, and the relative positional relationship between the metaphase chromosome karyotype and the image.

[0110] Establish a spatial coordinate system transformation algorithm for switching between different magnification objectives to obtain the estimated coordinate position of the kernel under a 100x objective lens; based on the physical coordinates (O) of the center of the sample slice image. x O y ), the pixel coordinates of the center of the target chromosome karyotype region in the sample slice image (S) x S y The physical width V of the sample slice image x and height V y By combining the horizontal pixel count W and vertical pixel count H of the sample slice image, the estimated physical coordinates of the center of the target chromosome karyotype region under a 100x objective lens can be calculated (A). xA y ):

[0111]

[0112] Specifically, the objective lens switching and sample slice displacement include the following steps:

[0113] Drive the motorized XY stage and motorized Z-axis back to zero, switch the current objective to 100x objective via the motorized objective converter, select any recorded target karyotype, and drive the motorized XY stage to move the target karyotype to the estimated coordinate position under the 100x objective.

[0114] Specifically, the extended grid imaging and target kernel cross-scale image matching include the following steps:

[0115] Autofocus is achieved with 100x field of view through coordinated control of the motorized Z-axis and camera, resulting in clear images under 100x objective lens.

[0116] A cross-scale image registration algorithm is used to match the same target keratin pattern images at different magnifications. It is used to determine whether there is a target keratin pattern at 100x magnification. If the target keratin pattern cannot be effectively identified, the grid imaging around the current field of view center is performed. The grid images are then stitched together and the target keratin pattern is matched.

[0117] Specifically, the calculation of the actual coordinate position of the kernel and the correction of the coordinate position deviation include the following steps:

[0118] Based on the image matching results and coordinate system transformation algorithm, and according to the estimated coordinate position under a 100x objective lens (A... x A y ), the pixel coordinates of the center of the target chromosome karyotype region in the stitched image (S x ', S y '), Physical width V of the sample slice image x 'and height V y 'And the number of horizontal pixels W' and vertical pixels H' of the stitched image, the actual physical coordinates of the center of the target chromosome karyotype region under a 100x objective lens can be calculated (B). x B y ):

[0119]

[0120] The actual coordinate position (B) x B y ) and estimated coordinate position (A) x A y By comparing the coordinates, the positional deviations in the X and Y directions can be calculated:

[0121] Δx'=Bx -A x

[0122] Δy'=B y -A y

[0123] Based on the coordinate position deviation, the electric XY stage is driven to precisely move the target kernel to the center of the field of view under the 100x objective lens.

[0124] Specifically, the high-magnification metaphase chromosome karyotype imaging includes the following steps:

[0125] Take microscopic images of the target karyotype at the center of the field of view;

[0126] Save the microscopic images in common image formats such as JPG and PNG.

[0127] Specifically, in the accompanying drawings, shapes and dimensions may be enlarged for clarity, and the same reference numerals will be used in all figures to indicate the same or similar parts.

[0128] In this embodiment, Figure 4 It is a chromosome karyotype microscopic scanning imaging device, in which 11 is a camera, 12 is an optical path correction system, 13 is an electric objective lens converter, 14 is a 10x objective lens, 15 is a 100x objective lens, 16 is an electric XY stage, 17 is a chromosome sample slide, and 18 is an instrument base.

[0129] Figure 5 This is a schematic diagram of the positional relationship of chromosome karyotype images under a 10x objective lens. In the diagram, 17 is a chromosome sample slice, 21 is a 10x microscopic image, and 22 is the karyotype of chromosomes in metaphase of the region of interest (ROI).

[0130] Figure 6 This is a schematic diagram of the calculation of coordinate positions for chromosome karyotype estimation under a 100x objective lens. In the diagram, 16 is the motorized XY stage, 17 is the chromosome sample slice, and 18 is the instrument base.

[0131] Figure 7 This is a schematic diagram of the expanded grid image under a 100x objective lens, where 31 is the current field of view center, 32 is the target kernel, and 33 is the expanded grid mosaic image.

[0132] A method for rapid search and precise localization of chromosome karyotypes includes the following steps:

[0133] Low-magnification whole-section scanning of chromosome sample: Using a 10x objective lens 14, the whole section of chromosome sample 17 was scanned to obtain overall 10x microscopic image 21 data;

[0134] Metaphase chromosome karyotype target detection and karyotype coordinate position recording: The target detection model (e.g., Faster-RCNN) is used to identify the ROI metaphase chromosome karyotype 22 in the 10x magnification microscopic image 21 obtained by scanning, mark the key information such as the target metaphase chromosome karyotype region, and record the coordinate position of all karyotypes;

[0135] Spatial coordinate system transformation and karyotype estimation coordinate position calculation: Establish a spatial coordinate system for the physical spatial relationship between the motorized XY stage 16, chromosome sample slice 17, 10x microscopic image 21, and ROI metaphase chromosome karyotype 22. Obtain the estimated coordinate position of the karyotype under 100x objective lens 15 according to the spatial coordinate system transformation algorithm under different magnification objectives.

[0136] Objective lens switching and sample displacement: Based on the recognition results and coordinate transformation relationship, switch the objective lens from 10x 14 to 100x 15, and at the same time place the target kernel 32 under the field of view of the 100x objective lens 15 according to the estimated coordinate position;

[0137] Extended grid imaging and cross-scale image matching of target karyotype: After completing the objective lens switching and sample displacement, the chromosome sample is imaged using a 100x objective lens 15. The same target karyotype 32 is matched at different magnifications using a SIFT-based cross-scale image matching algorithm. If the target karyotype 32 is not matched, extended grid imaging is performed with the current field of view center 31. The grid is extended and stitched together 33 and then the target karyotype 32 is matched until the target karyotype 32 is successfully matched.

[0138] Calculation of actual coordinate position of karyotype and correction of coordinate position deviation: Based on the matching results of karyotype 22 of ROI in metaphase and the coordinate system transformation algorithm, the actual coordinate position of the target karyotype 32 under 100x objective lens 15 is calculated. The actual coordinate position of the target karyotype 32 under 100x objective lens 15 is compared with the estimated coordinate position. The deviation in the X and Y directions is calculated. Then, the electric XY stage 16 is driven to move the target karyotype 32 to the center of the field of view 31.

[0139] High-magnification metaphase chromosome karyotype imaging: The target karyotype 32 at the center of the field of view 31 under a 100x objective lens 15 is photographed, and the 10x microscopic image 21 is saved as a common image format such as jpg or png.

[0140] S100: Low-power chromosome sample whole-section image scanning. S100 specifically includes the following steps:

[0141] The sample slice 17 containing the chromosome karyotype is placed on the motorized XY stage 16, ensuring that the sample slice is flat and firmly fixed to avoid displacement or shaking during subsequent operations. Then, the motorized biological microscope is fully initialized, including adjusting the light source intensity, calibrating the lens parameters, and initializing the stage.

[0142] Switching to the 10x objective lens 14, and using precise control of the motorized XY stage 16, the chromosome sample slice 17 is smoothly transferred to the pre-set initial scanning position. Then, the system activates autofocus, enabling the camera 11 to capture a clear 10x microscopic image 21 within a single field of view.

[0143] Under the coordinated exposure of the LED light source, after ensuring that the 10x microscopic image 21 is uniformly exposed and free of shadows, the full-section automatic scanning program is started according to the pre-planned scanning path. During the scanning process, the microscope continuously acquires 10x microscopic images 21, ensuring that each area is precisely covered, thereby obtaining a series of high-quality 10x microscopic images 21.

[0144] S200: Metaphase chromosome karyotype target detection and karyotype coordinate location recording specifically includes the following steps:

[0145] Based on the Faster-RCNN target detection model, target detection of mid-phase chromosome karyotype in all 10x microscopic images was performed on ROI 22.

[0146] Identify metaphase chromosome karyotypes of ROIs with research value: The main characteristics are that the chromosome karyotypes are elongated, highly dispersed, and almost non-overlapping;

[0147] Based on these characteristics, we can quickly and accurately search for and locate the metaphase chromosome karyotype of the ROI that meets the criteria.

[0148] If a matching ROI karyotype 22 is found, its precise coordinates in the 10x microscopic image 21 are calculated and recorded. If no matching karyotype is found in the current 10x microscopic image 21, the system automatically switches to the next acquired 10x microscopic image 21 to continue detection and analysis.

[0149] S300: Spatial coordinate system transformation and kernel estimation coordinate position calculation specifically include the following steps:

[0150] Establish a spatial coordinate positioning system S300-1: Establish a spatial coordinate positioning system under two different magnification objectives, which covers the spatial positioning relationship in four dimensions;

[0151] The spatial positioning relationship in the four dimensions specifically includes the following steps:

[0152] Positional relationship of the electric XY stage 16 relative to the instrument base S300-1a: defines the precise position of the electric XY stage 16 within its mechanical frame;

[0153] The positional relationship of chromosome sample slice 17 relative to the motorized XY stage S300-1b: Since chromosome sample slice 17 is fixed on motorized XY stage 16, its positional range is definite and limited;

[0154] Positional relationship between 10x microscopic image 21 and chromosome sample slice 17 S300-1c: describes the correspondence between 10x microscopic image 21 captured under a microscope and actual chromosome sample slice 17;

[0155] Positional relationship of ROI metaphase chromosome karyotype 22 relative to 10x microscopic image 21 S300-1d: The specific location of ROI metaphase chromosome karyotype 22 in the captured 10x microscopic image 21 was precisely determined;

[0156] The estimated coordinate position (A) of the nucleus under a 100x objective lens 15 was obtained using the S300-2 spatial coordinate system transformation algorithm under different magnification objectives. x A y S300-2a;

[0157]

[0158] in:

[0159] A x Estimate the physical coordinates of the target point along the X-axis at 10x magnification; A y Estimate the physical coordinates of the target point along the Y-axis at 10x magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; S x The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; S y The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; V x V represents the physical width of the sample slice image. y is the physical height of the sample slice image; W is the horizontal pixel count of the sample slice image; H is the vertical pixel count of the sample slice image.

[0160] The physical coordinates (O) of the center of the sample slice image x O y The karyotype region of the target chromosome changes with the scanning displacement, and the origin is the lower left corner (0,0) pixel of the sample slice image in which it is located.

[0161] S400: Objective lens conversion and sample slice displacement specifically include the following steps:

[0162] The estimated coordinate position of the kernel type under the 100x objective lens 15 is obtained by the spatial coordinate system transformation algorithm under different magnification objectives. The electric Z-axis is driven to return to zero. The current objective lens is switched to the 100x objective lens 15 through the electric objective lens converter 13. The electric XY stage 16 is driven to place a target kernel type 32 under the field of view of the 100x objective lens 15 according to the estimated coordinate position.

[0163] S500: Extended mesh imaging and target kernel cross-scale image matching specifically include the following steps:

[0164] Autofocus is achieved with 100x field of view through coordinated control of the motorized Z-axis and camera 11, resulting in a clear image under 100x objective lens 15;

[0165] A SIFT-based cross-scale image registration algorithm is used to match the same target keratin 32 images at different magnifications to determine whether the target keratin is present at 100x magnification. If the target keratin 32 cannot be effectively identified, the surrounding grid imaging is performed with the current field of view center, and the grid images are stitched together before the target keratin 32 image is matched.

[0166] Calculating the position coordinates S300-2b of the grid point specifically includes the following steps:

[0167] Scan step size calculation: Let the width of the rectangular ROI region be w, the height be h, and the overlap coefficient be α. Then the scan step size is:

[0168]

[0169] Where Δx is the step size in the X direction and Δy is the step size in the Y direction;

[0170] The coordinates of the scan grid points are generated by creating a 3×3 scan matrix centered at the initial point (x0, y0). The coordinates of each scan grid point are as follows:

[0171]

[0172] Where i,j∈{0,1,2}, corresponding to 9 scan points:

[0173] S={(x ij ,y ij )∣i,j=0,1,2}

[0174] The total coverage area size of the extended grid imaging mosaic is:

[0175]

[0176] α is the overlap coefficient between adjacent fields of view: α∈(0,1); Δx is the displacement of the stage in the X direction; Δy is the displacement of the stage in the Y direction; W″ is the total width of the stitched image; H″ is the total height of the stitched image.

[0177] α is the overlap coefficient between adjacent fields of view: α∈(0,1); Δx is the displacement of the motorized XY stage in the X direction; Δy is the displacement of the motorized XY stage in the Y direction; W″ is the total width of the stitched image 33; H″ is the total height of the stitched image 33.

[0178] S600: The calculation of actual coordinates of the kernel and the correction of coordinate position deviation specifically include the following steps:

[0179] The actual coordinates (B) of target nucleus 32 obtained by extended grid imaging under 100x objective lens 15. x B y ):

[0180]

[0181]

[0182] Among them, B x B represents the actual physical coordinates of the target point along the X-axis under a 100x objective lens. y A represents the actual physical coordinates of the target point along the Y-axis under a 100x objective lens; x Estimate the physical coordinates along the X-axis under a 100x objective lens; A y Estimate the physical coordinates along the Y-axis under a 100x objective lens; S x ′ represents the X-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; S y ′ represents the Y-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; V x ′ represents the physical width of the stitched images; V y ′ represents the physical height of the stitched image; W′ represents the horizontal pixel count of the stitched image; H′ represents the vertical pixel count of the stitched image.

[0183] The actual coordinates (B) of target nucleus 32 obtained by extended grid imaging under 100x objective lens 15. x B y ) and estimated coordinate position (A) x A y By comparing the positions in the X and Y directions, the positional deviations can be calculated.

[0184] Δx'=B x -A x

[0185] Δy'=B y -A y

[0186] Drive the electric XY stage 16 based on the positional deviation to precisely move the target kernel 32 to the center of the field of view 31 under the 100x objective lens 15, and obtain the final coordinate position S300-2c.

[0187] S700: High-power metaphase chromosome karyotype imaging specifically includes the following steps:

[0188] Take a picture of the target kernel image at the center of the field of view;

[0189] Save the image as a common image format such as JPG or PNG.

[0190] The present invention also provides a system for searching and locating chromosome karyotypes. The system for searching and locating chromosome karyotypes can be implemented by executing the process steps of the method for searching and locating chromosome karyotypes. That is, those skilled in the art can understand the method for searching and locating chromosome karyotypes as a preferred embodiment of the system for searching and locating chromosome karyotypes.

[0191] A system for searching and locating chromosome karyotypes according to the present invention includes:

[0192] Module M1: Scans chromosome samples and acquires microscopic images under a low-power objective lens;

[0193] Module M2: Based on the microscopic image, detect and record the coordinate position of the chromosome karyotype;

[0194] Module M3: Establishes a spatial coordinate system to obtain the estimated coordinate position of the chromosome karyotype under a high-power objective lens;

[0195] Module M4: Under high magnification, move the target chromosome karyotype to the estimated coordinate position;

[0196] Module M5: Performs image matching of the same target karyotype at different magnifications and determines whether the matching is successful. If the result is yes, the image matching result is obtained, and module M6 is triggered; if the result is no, extended grid imaging is performed with the current field of view center, the grid image is stitched together, and module M5 is triggered again until the grid image of the target chromosome karyotype is successfully matched.

[0197] Module M6: Based on the image matching results, calculate the actual coordinate position of the target chromosome karyotype, then determine the deviation between the actual coordinate position and the estimated coordinate position, move the target chromosome karyotype to the center of the field of view; acquire and save the image of the target chromosome karyotype.

[0198] Preferably, in module M2, the coordinate positions of the metaphase chromosome karyotype are detected and recorded using the Faster-RCNN object detection model.

[0199] Preferably, in module M3, the estimated coordinate position of the chromosome karyotype is expressed mathematically as follows:

[0200]

[0201] Among them, A x Estimate the physical coordinates of the target point along the X-axis under low magnification; A y Estimate the physical coordinates of the target point along the Y-axis under low magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; S x The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; S y The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; V x V represents the physical width of the sample slice image. y is the physical height of the sample slice image; W is the horizontal pixel count of the sample slice image; H is the vertical pixel count of the sample slice image.

[0202] Preferably, in module M5, the SIFT algorithm is used to match the image with the target chromosome karyotype at different magnifications;

[0203] The stitched grid image includes:

[0204] Module A1: Calculates the scan step distance and generates scan grid points; the scan grid points are initialized at (x0, y0) to generate a 3×3 scan matrix;

[0205] Module A2: Stitching and expanding grid imaging;

[0206] In module A1, the mathematical expression for the scanning step distance is:

[0207]

[0208] Where Δx is the step size in the X direction, Δy is the step size in the Y direction; w is the width of the rectangular ROI region; h is the height of the rectangular ROI region; α is the overlap coefficient; the symbol · represents the product;

[0209] The mathematical expression for the coordinates of the scanned grid points is:

[0210]

[0211] Where, x ij Let y be the x-coordinate of the scanned grid point. ijThe ordinate of the scan grid point is (x0, y0); (x0, y0) is the initial point, i.e., the center; i and j are the grid index positions used to identify the specific positions, i, j∈{0,1,2};

[0212] In module A2, the total coverage area size of the extended grid imaging is mathematically expressed as:

[0213]

[0214] α is the overlap coefficient between adjacent fields of view: α∈(0,1); Δx is the displacement of the stage in the X direction; Δy is the displacement of the stage in the Y direction; W″ is the total width of the stitched image; H″ is the total height of the stitched image.

[0215] Preferably, in module M6, the mathematical expression for the actual coordinate position is:

[0216]

[0217] Among them, B x B represents the actual physical coordinates of the target point along the X-axis under high magnification; y A represents the actual physical coordinates of the target point along the Y-axis under high magnification. x To estimate the physical coordinates along the X-axis under high magnification; A y To estimate the physical coordinates along the Y-axis under high magnification; S x ′ represents the X-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; S y ′ represents the Y-pixel coordinate of the center of the target chromosome karyotype region in the stitched image; V x ′ represents the physical width of the stitched images; V y ′ represents the physical height of the stitched image; W′ represents the horizontal pixel count of the stitched image; H′ represents the vertical pixel count of the stitched image.

[0218] The mathematical expression for the deviation between the actual coordinate position and the estimated coordinate position is:

[0219] Δx'=B x -A x

[0220] Δy'=B y -A y

[0221] Where Δx' is the deviation in the X-axis direction and Δy' is the deviation in the Y-axis direction.

[0222] Those skilled in the art will understand that, besides implementing the system and its various devices, modules, and units provided by this invention in the form of purely computer-readable program code, the same functions can be achieved entirely through logical programming of the method steps, making the system and its various devices, modules, and units of this invention function in the form of logic gates, switches, application-specific integrated circuits, programmable logic controllers, and embedded microcontrollers. Therefore, the system and its various devices, modules, and units provided by this invention can be considered as a hardware component, and the devices, modules, and units included therein for implementing various functions can also be considered as structures within the hardware component; alternatively, the devices, modules, and units for implementing various functions can be considered as both software modules implementing the method and structures within the hardware component.

[0223] In the description of this application, it should be understood that the terms "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application.

[0224] Specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art can make various changes or modifications within the scope of the claims, which do not affect the essence of the present invention. Unless otherwise specified, the embodiments and features described in this application can be arbitrarily combined with each other.

Claims

1. A method for searching and locating chromosome karyotypes, characterized in that, include: Step S1: Scan the chromosome sample and acquire microscopic images under a low-power objective lens; Step S2: Based on the microscopic image, detect and record the coordinate position of the chromosome karyotype; Step S3: Establish a spatial coordinate system, and then obtain the estimated coordinate position of the chromosome karyotype under a high-power objective lens; Step S4: Under a high-power objective lens, move the target chromosome karyotype to the estimated coordinate position; Step S5: Perform image matching of the same target karyotype at different magnifications and determine whether the matching is successful; if the result is yes, obtain the image matching result and proceed to step S6; if the result is no, perform extended grid imaging with the current field of view center, stitch the grid images, and re-execute step S5 until the grid image of the target chromosome karyotype is successfully matched. Step S6: Based on the image matching results, calculate the actual coordinate position of the target chromosome karyotype, then determine the deviation between the actual coordinate position and the estimated coordinate position, and move the target chromosome karyotype to the center of the field of view; Acquire and save images of the target chromosome karyotype; In step S5, the SIFT algorithm is used to match the target chromosome karyotype at different magnifications. The process of stitching the grid images includes: Step A1: Calculate the scan step distance and generate scan grid points; the scan grid points are defined as ( , Using 1 as the initial point, generate a 3×3 scan matrix; Step A2: Stitch and expand the grid imaging; In step A1, the mathematical expression for the scanning step distance is: in, The step size in the X direction. The step size in the Y direction; The width of the rectangular ROI region; This represents the height of the rectangular ROI region. The overlap coefficient; symbol Indicates product; The mathematical expression for the coordinates of the scanned grid points is: in, The x-coordinate of the scanned grid points. The vertical coordinates of the scanned grid points; , () is the initial point, i.e., the center. and This is the index position of the grid, used to identify a specific location. , ∈{0,1,2}; In step A2, the total coverage area size of the expanded grid imaging is mathematically expressed as: The overlap coefficient between adjacent fields of view: ∈(0,1); This represents the displacement of the stage in the X direction; This represents the displacement of the stage in the Y direction; This represents the total width of the stitched image; This represents the total height of the stitched image.

2. The method for searching and locating chromosome karyotypes according to claim 1, characterized in that, In step S2, the coordinates of the metaphase chromosome karyotype are detected and recorded using the Faster-RCNN object detection model.

3. The method for searching and locating chromosome karyotypes according to claim 1, characterized in that, In step S3, the estimated coordinate position of the chromosome karyotype is expressed mathematically as follows: in, Estimate the physical coordinates of the target point along the X-axis under low magnification; Estimate the physical coordinates of the target point along the Y-axis under low magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; The physical width of the sample slice image; The physical height of the sample slice image; The number of horizontal pixels in the sample slice image; The vertical pixel count of the sample slice image.

4. The method for searching and locating chromosome karyotypes according to claim 1, characterized in that, In step S6, the mathematical expression for the actual coordinate position is: in, The actual physical coordinates of the target point along the X-axis under high magnification objective lens; The actual physical coordinates of the target point along the Y-axis under high magnification objective lens; To estimate the physical coordinates along the X-axis under high-magnification objectives; To estimate the physical coordinates along the Y-axis under high-magnification objectives; The X-pixel coordinates of the center of the target chromosome karyotype region in the stitched image; The Y-pixel coordinates of the center of the target chromosome karyotype region in the stitched image; The physical width of the stitched images; The physical height of the stitched images; This represents the number of horizontal pixels in the stitched image. This represents the vertical pixel count of the stitched images. The mathematical expression for the deviation between the actual coordinate position and the estimated coordinate position is: in, This represents the deviation in the X-axis direction; This represents the deviation in the Y-axis direction.

5. A system for searching and locating chromosome karyotypes, characterized in that, include: Module M1: Scans chromosome samples and acquires microscopic images under a low-power objective lens; Module M2: Based on the microscopic image, detect and record the coordinate position of the chromosome karyotype; Module M3: Establishes a spatial coordinate system to obtain the estimated coordinate position of the chromosome karyotype under a high-power objective lens; Module M4: Under high magnification, move the target chromosome karyotype to the estimated coordinate position; Module M5: Performs image matching of the same target kernel type at different magnifications and determines whether the matching is successful; if the result is successful, the image matching result is obtained, and module M6 is triggered to work; If the result is negative, then expand the grid imaging at the current field of view center, stitch the grid images, and re-trigger module M5 until the grid image of the target chromosome karyotype is successfully matched. Module M6: Based on the image matching results, calculate the actual coordinate position of the target chromosome karyotype, then determine the deviation between the actual coordinate position and the estimated coordinate position, and move the target chromosome karyotype to the center of the field of view; Acquire and save images of the target chromosome karyotype; In module M5, the SIFT algorithm is used to match the image with the target chromosome karyotype at different magnifications; The stitched grid image includes: Module A1: Calculates the scan step distance and generates scan grid points; the scan grid points are defined as ( , Using 1 as the initial point, generate a 3×3 scan matrix; Module A2: Stitching and expanding grid imaging; In module A1, the mathematical expression for the scanning step distance is: in, The step size in the X direction. The step size in the Y direction; The width of the rectangular ROI region; This represents the height of the rectangular ROI region. The overlap coefficient; symbol Indicates product; The mathematical expression for the coordinates of the scanned grid points is: in, The x-coordinate of the scanned grid points. The vertical coordinates of the scanned grid points; , () is the initial point, i.e., the center. and This is the index position of the grid, used to identify a specific location. , ∈{0,1,2}; In module A2, the total coverage area size of the extended grid imaging is mathematically expressed as: The overlap coefficient between adjacent fields of view: ∈(0,1); This represents the displacement of the stage in the X direction; This represents the displacement of the stage in the Y direction; This represents the total width of the stitched image; This represents the total height of the stitched image.

6. The system for searching and locating chromosome karyotypes according to claim 5, characterized in that, In module M2, the coordinates of the metaphase chromosome karyotype are detected and recorded using the Faster-RCNN object detection model.

7. The system for searching and locating chromosome karyotypes according to claim 5, characterized in that, In module M3, the estimated coordinate position of the chromosome karyotype is expressed mathematically as follows: in, Estimate the physical coordinates of the target point along the X-axis under low magnification; Estimate the physical coordinates of the target point along the Y-axis under low magnification; O x The X physical coordinates of the center of the sample slice image; O y The physical Y coordinate of the center of the sample slice image; The X-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; The Y-pixel coordinates of the center of the target chromosome karyotype region in the sample slice image; The physical width of the sample slice image; The physical height of the sample slice image; The number of horizontal pixels in the sample slice image; The vertical pixel count of the sample slice image.

8. The system for searching and locating chromosome karyotypes according to claim 5, characterized in that, In module M6, the mathematical expression for the actual coordinate position is: in, The actual physical coordinates of the target point along the X-axis under high magnification objective lens; The actual physical coordinates of the target point along the Y-axis under high magnification objective lens; To estimate the physical coordinates along the X-axis under high-magnification objectives; To estimate the physical coordinates along the Y-axis under high-magnification objectives; The X-pixel coordinates of the center of the target chromosome karyotype region in the stitched image; The Y-pixel coordinates of the center of the target chromosome karyotype region in the stitched image; The physical width of the stitched images; The physical height of the stitched images; This represents the number of horizontal pixels in the stitched image. This represents the vertical pixel count of the stitched images. The mathematical expression for the deviation between the actual coordinate position and the estimated coordinate position is: in, This represents the deviation along the X-axis. This represents the deviation in the Y-axis direction.

Images