Microscopic scanning system and microscopic scanning method

Abstract

The present disclosure provides a microscopic scanning system and a microscopic scanning method. The microscopic scanning system includes a glass slide carrier, a vacuum adsorption device, a light source and a microscopic camera system; the vacuum adsorption device is arranged on the glass slide carrier to adsorb the upper surface of the calibration film through the vacuum adsorption device, and the distance between the upper surface of the calibration film and the glass slide carrier is fixed; the light source is located below the glass slide carrier for providing light; the microscopic camera system is located above the glass slide carrier, and the light emitted by the light source passes through the calibration film and then irradiates the microscopic camera system to obtain an image of the calibration film, and obtains the focus reference coordinates according to the image of the calibration film. When the microscopic scanning system of the present invention leaves the factory, the calibration film is vacuum adsorbed to determine the focus reference point of the glass slide of each device, which greatly reduces the focus range in the automatic focusing process, realizes rapid fitting of the microscopic morphology of the glass slide, and thus realizes rapid scanning.

Description

Technical Field

[0001] The present disclosure relates to the technical field of automatic bright field microscopic scanning, and in particular to a microscopic scanning system and a microscopic scanning method. Background Art

[0002] Slide sample testing is a commonly used test item in modern medicine. Automatic bright-field microscope scanners have gradually replaced manual scanning methods and have been widely used due to their advantages such as high stability and fast detection efficiency.

[0003] In order to improve the scanning speed of the automatic bright field microscope scanner and determine the scanning reference, it is necessary to obtain the microscopic morphology of the slide. The currently commonly used method is to print multiple reference marking points on the slide or cover glass, obtain the Z-axis coordinates of the multiple marking points through the automatic focusing method, and fit the microscopic morphology of the upper surface of the slide according to the multiple Z-axis coordinates to determine the scanning reference focal surface.

[0004] Prior art CN113302539B discloses a method for improving microscopic imaging, which calculates a reference focal plane defining the surface of a cover glass or a slide glass by identifying a plurality of reference marking points printed on the surface of a cover glass or a slide glass used in combination with an optical microscope. This method can only be applied to thin samples or when the marking points do not overlap with the sample. When the sample is thick or the sample and the reference marking point overlap in height, the sample and the reference marking point cannot be effectively distinguished.

[0005] Prior art CN111240001B discloses a focusing method for a microscopic specimen with a cover glass, which prints reference marking points on the cover glass, focuses on the reference marking points on the cover glass, obtains the focal plane positions of multiple reference marking points, fits a reference plane based on these focal plane positions, and estimates the focal plane position of the sample based on the reference plane. This method still has the problem that when the sample is thick or the sample and the reference mark overlap in the height direction, it cannot effectively distinguish between the sample and the reference mark; when the marking points are distributed at the four corners of the cover glass, the marking points are far away from the sample, and the fitted plane is quite different from the actual curved surface of the sample, resulting in distortion of the scanned image.

[0006] In addition, due to the different thicknesses of different glass slides or the deformation of the same glass slide caused by clamping, when finding the focal position of the reference mark point, each reference mark point needs to be focused over a large range, resulting in a long scanning process.

[0007] In summary, the current scanning method based on printing reference points on a slide to determine the reference focal plane has the following problems: ① The marking point and the sample area cannot overlap in the Z-axis direction, otherwise the sample and the marking point cannot be effectively distinguished, resulting in failure of fitting the focal surface, loss of focus during the scanning process, and poor scanning stability; ② In order to avoid the overlap of the marking points with the sample, the marking points are all at the four corners of the slide, far away from the sample area, resulting in a large difference between the fitted slide micromorphology and the slide micromorphology in the sample area, and poor fitting accuracy; ③ There should be no oil or impurities above the marking point, otherwise it will be impossible to effectively distinguish the marking point from the impurities, resulting in failure in fitting the focal surface; ④ Due to the different thicknesses of different glass slides, or the deformation of the same glass slide when clamped, it is necessary to focus on a large range of each marking point during the process of finding the marking point, which makes the scanning process too long. Summary of the invention

[0008] In order to solve at least one of the above technical problems, the present disclosure provides a microscopic scanning system and a microscopic scanning method.

[0009] According to one aspect of the present disclosure, there is provided a microscopic scanning system, comprising: Slide carrier; A vacuum adsorption device, the vacuum adsorption device is arranged on the glass slide carrier, so as to adsorb the upper surface of the calibration sheet through the vacuum adsorption device, and make the distance between the upper surface of the calibration sheet and the glass slide carrier fixed; a light source, located below the slide carrier and configured to provide light; and The microscope camera system is located above the slide carrier. The light emitted by the light source passes through the calibration film and then irradiates the microscope camera system to obtain an image of the calibration film, and the focus reference coordinates are obtained based on the image of the calibration film.

[0010] According to the microscope scanning system of at least one embodiment of the present disclosure, the vacuum adsorption device includes at least three vacuum suction cup assemblies; a through hole is opened on the glass slide carrier, and the at least three vacuum suction cup assemblies are respectively located at the corners of the through hole of the glass slide carrier, and the calibration piece is adsorbed by the vacuum suction cup assembly so that the upper surface of the calibration piece is fixed relative to the glass slide carrier.

[0011] According to the microscopic scanning system of at least one embodiment of the present disclosure, the vacuum chuck assembly includes: A base, wherein a gas circulation channel is provided at the upper end of the base; a receiving space is formed at the lower end of the base, the upper surface of the receiving space is formed as a positioning surface, and positioning beads are arranged in the receiving space; an adsorption hole is also provided at the lower end of the base, the adsorption hole is located at the periphery of the receiving space, and the adsorption hole is connected with the gas circulation channel; when the calibration piece is adsorbed by the adsorption hole of the vacuum suction cup assembly, the positioning beads are in contact with the upper surface of the calibration piece and the positioning surface respectively.

[0012] According to the microscopic scanning system of at least one embodiment of the present disclosure, the vacuum chuck assembly further includes: The elastic component is disposed on the base, and the lower end of the elastic component extends out of the lower end of the base.

[0013] According to another aspect of the present disclosure, there is provided a microscopic scanning method, comprising: Obtaining the focus reference coordinates of the microscanning system; Placing a glass slide on a glass slide carrier, and allowing the upper surface of the glass slide to be adsorbed and positioned by a vacuum adsorption device; wherein a plurality of first marking portions are provided on the upper surface of the glass slide, and an invisible light cutoff coating film is provided at the first marking portions; Obtaining N sampling points of the sample according to the position of the sample on the slide, wherein N is an integer greater than or equal to 1; Control the movement of the slide carrier and make the microscope camera system located at the focus reference coordinate directly above the i-th sampling point; control the microscope camera system to move in the height direction, and during the movement of the microscope camera system, take a plurality of non-visible light images including the first identification portion; wherein i is greater than or equal to 1 and less than or equal to N; According to the clarity of the multiple non-visible light images taken by the microscope camera system, a Z-axis coordinate Mi corresponding to the non-visible light image with the largest clarity including the first identification portion is obtained; According to a plurality of Z-axis coordinates Mi corresponding to a plurality of sampling points, a lower scanning reference plane is obtained; A multi-layer scan is performed above the lower scanning reference plane to obtain a plurality of intermediate images, which are then fused to obtain a scanned image.

[0014] According to the microscopic scanning method of at least one embodiment of the present disclosure, the focusing reference coordinates of the microscopic scanning system are obtained by reading the parameters of the microscopic scanning system; wherein the parameters of the microscopic scanning system are obtained by calibrating the microscopic scanning system.

[0015] According to at least one embodiment of the present disclosure, the microscopic scanning method calibrates the microscopic scanning system, including: Placing the calibration sheet on the slide carrier, and allowing the upper surface of the calibration sheet to be adsorbed and positioned by the vacuum adsorption device; wherein the calibration sheet includes a second identification portion; Controlling the movement of the slide carrier so that the microscope camera system is located directly above the second identification portion; controlling the microscope camera system to move in a height direction, and capturing a plurality of images including the second identification portion during the movement of the microscope camera system; The focus reference coordinates are obtained according to the Z-axis coordinate of the image with the highest definition among the multiple images obtained by the microscope camera system during the movement.

[0016] According to at least one embodiment of the present disclosure, a microscopic scanning method is provided on the glass slide, and the sample is located between the glass slide and the cover glass, wherein a plurality of third identification portions are provided on the lower surface of the cover glass, and an invisible light cutoff coating is provided at the third identification portions; the microscopic scanning method comprises: The upper scanning reference plane is obtained through the third identification part of the cover glass; when the microscopic scanning system is performing a scanning operation, multi-layer scanning is performed at a position between the lower scanning reference plane and the upper scanning reference plane.

[0017] According to the microscopic scanning method of at least one embodiment of the present disclosure, the third identification portion of the cover glass is different in shape from the first identification portion of the slide glass.

[0018] According to the microscopic scanning method of at least one embodiment of the present disclosure, after the glass slide is positioned by the vacuum adsorption device, the upper surface of the glass slide is located at the focus reference coordinates.

[0019] The technical solution of the present invention has the following beneficial effects: The microscopic scanning method disclosed in the present invention solves the problems that the marking points (the first marking part and the third marking part) cannot overlap with the sample area in the Z-axis direction and there cannot be oil stains or impurity interference above the marking points by taking pictures with an invisible light cutoff coating in conjunction with a microscopic camera system (a visible light camera with a partial invisible light collection function). The stability of marking point collection is improved, and the selected marking points are just below / above the sample. The fitted microscopic morphology of the slide / cover glass is more realistic and accurate. The calibration film is vacuum-adsorbed to determine the focus reference point of the slide of each device, which greatly reduces the focus range during the autofocus process and enables rapid fitting of the microscopic morphology of the slide / cover glass, thereby achieving rapid scanning. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The accompanying drawings illustrate exemplary embodiments of the present disclosure and together with the description serve to explain the principles of the present disclosure. These drawings are included to provide a further understanding of the present disclosure and are incorporated in and constitute a part of this specification.

[0021] Figure 1 It is a schematic structural diagram of a microscopic scanning system according to one embodiment of the present disclosure.

[0022] Figure 2 It is a schematic structural diagram of a vacuum suction cup assembly according to one embodiment of the present disclosure.

[0023] Figure 3 It is a schematic structural diagram of a vacuum suction cup assembly from another angle according to an embodiment of the present disclosure.

[0024] Figure 4 is a flow chart of a microscopic scanning method according to one embodiment of the present disclosure.

[0025] Figure 5 is a schematic structural diagram of a glass slide according to one embodiment of the present disclosure.

[0026] Figure 6 is a schematic structural diagram of a cover glass according to one embodiment of the present disclosure.

[0027] Description of Reference Numerals 10 Slide Carrier 20 Vacuum adsorption device 21 base 22 Positioning surface 23 Positioning beads 24 adsorption holes 25 Elastic components 30 Light Source 40 Microscope camera system 50 slides 51 First Logo Department 60 Calibration film 61 Second Logo Section 70 Cover glass 71 Third Logo Section. DETAILED DESCRIPTION

[0028] The present disclosure is further described in detail below in conjunction with the accompanying drawings and implementations. It is understood that the specific implementations described herein are only used to explain the relevant content, rather than to limit the present disclosure. It should also be noted that, for ease of description, only the parts related to the present disclosure are shown in the accompanying drawings.

[0029] It should be noted that, in the absence of conflict, the embodiments and features of the embodiments in the present disclosure can be combined with each other. The technical solution of the present disclosure will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.

[0030] Unless otherwise specified, the exemplary embodiments / embodiments shown will be understood as providing exemplary features of various details of some ways in which the technical concept of the present disclosure can be implemented in practice. Therefore, unless otherwise specified, the features of the various embodiments / embodiments can be combined, separated, interchanged and / or rearranged without departing from the technical concept of the present disclosure.

[0031] The use of cross-hatching and / or shading in the accompanying drawings is generally used to make the boundaries between adjacent components clear. As such, unless otherwise specified, the presence or absence of cross-hatching or shading does not convey or indicate any preference or requirement for the specific materials, material properties, dimensions, proportions, commonalities between the components shown, and / or any other characteristics, attributes, properties, etc. of the components. In addition, in the accompanying drawings, the sizes and relative sizes of the components may be exaggerated for clarity and / or descriptive purposes. When the exemplary embodiments can be implemented differently, the specific process sequence can be performed in a different order than described. For example, two successively described processes can be performed substantially simultaneously or in an order opposite to the described order. In addition, the same figure numbers represent the same components.

[0032] The terms used herein are for the purpose of describing specific embodiments, and are not intended to be restrictive. As used herein, unless the context clearly indicates otherwise, the singular forms "one (kind, person)" and "said (the)" are also intended to include plural forms. In addition, when the terms "comprise" and / or "include" and their variations are used in this specification, it is explained that there are stated features, integral bodies, steps, operations, parts, assemblies and / or their groups, but it is not excluded that there are or add one or more other features, integral bodies, steps, operations, parts, assemblies and / or their groups. It should also be noted that, as used herein, the terms "substantially", "approximately" and other similar terms are used as approximate terms and not as degree terms, so that they are used to explain the inherent deviations of the measured values, calculated values ​​and / or the values ​​provided that will be recognized by those of ordinary skill in the art.

[0033] Figure 1 It is a schematic structural diagram of a microscopic scanning system according to one embodiment of the present disclosure.

[0034] like Figure 1 As shown, the microscope scanning system of the present disclosure may include components such as a slide carrier 10, a vacuum adsorption device 20, a light source 30, and a microscope camera system 40.

[0035] In the microscopic scanning system disclosed in the present invention, the slide carrier 10, the light source 30 and the microscopic camera system 40 are all inherent structures in the microscopic scanning system, wherein the slide carrier 10 can be driven to produce two-dimensional motion in the horizontal direction, that is, the slide carrier 10 can move in the X direction and the Y direction, and the microscopic camera system 40 can be driven to produce motion in the Z direction. Of course, the slide carrier 10 disclosed in the present invention can also be driven to produce motion in the X direction, the Y direction and the Z direction, and the present invention does not limit the motion of these components.

[0036] The vacuum adsorption device 20 is disposed on the slide carrier 10, so that the upper surface of the calibration sheet 60 or the glass slide 50 is adsorbed by the vacuum adsorption device 20, and the distance between the upper surface of the calibration sheet 60 or the glass slide 50 and the glass slide carrier 10 is fixed. That is, when the microscopic scanning system of the present disclosure leaves the factory, the vacuum adsorption device 20 can adsorb the calibration sheet 60, and obtain the focus reference coordinate Z0 through the calibration sheet 60. When the microscopic scanning system of the present disclosure is used normally, the vacuum adsorption device 20 can adsorb the glass slide 50, and at this time, the glass slide 50 can be provided with a sample.

[0037] That is to say, when the microscopic scanning system of the present invention is in use, the distance between the upper surface of the glass slide 50 and the glass slide carrier 10 can be fixed by the setting of the vacuum adsorption device 20; moreover, the glass slide 50 will not deform or will produce a small deformation, thereby ensuring that the position of the upper surface of the glass slides 50 of different thicknesses is the same as the focusing reference coordinate Z0, and the use of the vacuum adsorption method will not cause friction between the glass slide 50 and the glass slide carrier 10, and will not produce glass residues to damage the equipment or user safety.

[0038] Accordingly, when the microscopic scanning system of the present disclosure is in autofocus, the autofocus time can be shortened. In addition, before leaving the factory, the microscopic scanning system absorbs the calibration sheet 60 (the calibration sheet 60 can be considered as a glass slide 50 with a special structure) and automatically focuses on the second identification portion 61 on the calibration sheet 60, thereby obtaining the Z-axis coordinate of the upper surface of the calibration sheet 60, which can be considered as the focus reference coordinate Z0 of the microscopic scanning system.

[0039] In one embodiment, the second identification portion 61 is formed as a positioning point on the calibration sheet 60. For example, only one second identification portion 61 may be provided on the calibration sheet 60, and at this time, the microscopic camera system 40 may be located directly above the second identification portion 61, so that the second identification portion 61 can be conveniently and quickly licensed and the focus reference coordinates of the microscopic scanning system can be obtained.

[0040] Generally speaking, the microscopic scanning system of the present disclosure only needs to be calibrated once before leaving the factory, that is, the position of the upper surface of the slide 50 can be known when the microscopic scanning system is working.

[0041] The light source 30 is located below the slide carrier 10 and is used to provide light. The light source 30 of the present disclosure can emit visible light and invisible light. Specifically, the invisible light emitted by the light source 30 of the present disclosure can be infrared light. In a preferred embodiment, the light source 30 can provide visible light and near-infrared light at different times.

[0042] The microscope camera system 40 is located above the slide carrier 10 . The light emitted by the light source 30 passes through the calibration film 60 and then irradiates the microscope camera system 40 to obtain an image of the calibration film 60 . Based on the image of the calibration film 60 , the focus reference coordinates are obtained.

[0043] In one embodiment, the microscopic imaging system 40 may include components such as an objective lens, a lens barrel, and a camera, and the microscopic imaging system 40 may be controlled by a motion mechanism to achieve movement of the microscopic imaging system. The camera included in the microscopic imaging system 40 may be capable of imaging visible light images (visible light images) and invisible light images (invisible light images). In other words, the camera is a visible light camera with a partial invisible light acquisition function. In a preferred embodiment, the camera may generate visible light images and infrared light images.

[0044] Refer to Figure 1 The vacuum adsorption device 20 disclosed in the present invention includes at least three vacuum suction cup assemblies; a through hole is opened on the glass slide carrier 10, and the at least three vacuum suction cup assemblies are respectively located at the corners of the through hole of the glass slide carrier 10.

[0045] In a specific embodiment, the through hole formed on the slide carrier 10 is a rectangular hole. In this case, the number of the vacuum suction cup assemblies is four, and the four vacuum suction cup assemblies are respectively arranged at the four corners of the rectangular hole. Those skilled in the art should know that the number of the vacuum suction cup assemblies disclosed in the present invention can also be set to other values, and the arrangement positions thereof can also vary with the different shapes of the through holes of the slide carrier 10.

[0046] In a preferred embodiment, the vacuum chuck assembly is arranged to determine the position of a plane, which is the upper surface of the slide 50. That is, all the vacuum chuck assemblies of the present disclosure cannot be arranged in a straight line.

[0047] Figure 2 It is a schematic structural diagram of a vacuum suction cup assembly according to one embodiment of the present disclosure. Figure 3 It is a schematic structural diagram of a vacuum suction cup assembly from another angle according to an embodiment of the present disclosure.

[0048] like Figure 2 As shown, the vacuum chuck assembly of the present disclosure may include a base 21, which can be fixed to the glass slide carrier 10. A gas flow channel is opened at the upper end of the base 21, and the gas flow channel can be connected to the vacuum pipeline, so that negative pressure can be provided to the vacuum chuck assembly.

[0049] A accommodating space is formed at the lower end of the base 21, and the upper surface of the accommodating space is formed as a positioning surface 22, and positioning beads 23 are arranged in the accommodating space; an adsorption hole 24 is also opened at the lower end of the base 21, and the adsorption hole 24 is located at the periphery of the accommodating space, and the adsorption hole 24 is connected to the gas flow channel; when the glass slide 50 is adsorbed by the adsorption hole 24 of the vacuum suction cup assembly, the positioning beads 23 contact the upper surface of the glass slide 50 and the positioning surface 22 respectively.

[0050] like Figure 3 As shown, the number of the adsorption holes 24 of the present invention may be 10, and the 10 adsorption holes 24 are evenly arranged around the periphery of the accommodating space, thereby, the vacuum suction cup assembly of the present invention can stably adsorb the glass slide 50 or the calibration sheet 60.

[0051] More preferably, the vacuum suction cup assembly further includes an elastic component 25, which is disposed on the base 21, and the lower end of the elastic component 25 extends to the outside (below) of the lower end of the base 21; that is, when the vacuum suction cup assembly of the present disclosure adsorbs the glass slide 50 or the calibration sheet 60, the elastic component 25 can directly contact the glass slide 50 or the calibration sheet 60, and the elastic component 25 forms a sealed space inside the elastic component 25 through the sealed contact between the elastic component 25 and the glass slide 50 or the calibration sheet 60. At this time, by applying negative pressure to the adsorption hole 24, the elastic component 25 is deformed, and the sealed space can always be in a sealed state. Accordingly, the glass slide 50 or the calibration sheet 60 will not fall off the vacuum suction cup assembly. In addition, due to the presence of the positioning beads 23, the upper surface of the glass slide 50 or the calibration sheet 60 can maintain a constant position relative to the base 21 of the vacuum suction cup assembly, and accordingly, it can also maintain a fixed distance relative to the glass slide carrier 10.

[0052] In a specific embodiment, the elastic component 25 of the present disclosure is formed as a cylindrical structure as a whole, and is arranged around the base 21, and its upper end can be fixed to the base 21. For example, the upper end of the elastic component 25 can be fixed to the upper end of the base 21 by bonding.

[0053] Figure 4 is a flow chart of a microscopic scanning method according to one embodiment of the present disclosure.

[0054] like Figure 4As shown, the microscopic scanning method disclosed in the present invention can be implemented based on the above-mentioned microscopic scanning system. Specifically, the microscopic scanning method disclosed in the present invention may include: S110, obtaining the focus reference coordinates of the microscopic scanning system; S120, placing the glass slide 50 on the glass slide carrier 10, and allowing the upper surface of the glass slide 50 to be adsorbed and positioned by the vacuum adsorption device 20; wherein the glass slide 50 includes a plurality of first identification portions 51, and an invisible light cutoff coating is provided at the first identification portion 51; S130, according to the position of the sample on the glass slide 50, obtaining N sampling points of the sample, wherein N is an integer greater than or equal to 1; S140, controlling the movement of the glass slide carrier 10, and allowing the microscopic camera system 40 to be located at the focus reference coordinates directly above the i-th sampling point; Control the microscope camera system 40 to move in the height direction, and during the movement of the microscope camera system 40, take a plurality of non-visible light images including the first identification portion 51; wherein, i is greater than or equal to 1 and less than or equal to N; S150, according to the clarity of the plurality of non-visible light images taken by the microscope camera system 40, obtain the Z-axis coordinate Mi corresponding to the non-visible light image including the first identification portion 51 with the highest clarity; S160, according to the plurality of Z-axis coordinates Mi corresponding to the plurality of sampling points, obtain a lower scanning reference plane; S170, perform multi-layer scanning above the lower scanning reference plane, and obtain a plurality of intermediate images, fuse the intermediate images, and obtain a scanned image.

[0055] The microscopic scanning method of the present invention will be described in detail below in conjunction with specific implementation forms.

[0056] S110, obtaining the focus reference coordinates of the microscopic scanning system.

[0057] In the present disclosure, the focusing reference coordinates of the microscopic scanning system are obtained by reading the parameters of the microscopic scanning system; wherein the parameters of the microscopic scanning system are obtained by calibrating the microscopic scanning system.

[0058] In other words, the microscopic scanning system of the present invention needs to be calibrated before leaving the factory, and the focus reference coordinates obtained in the calibration process are written into the memory of the microscopic scanning system. Therefore, when the microscopic scanning system of the present invention is in use, the focus reference coordinates can be directly read from the memory, thereby speeding up the focus speed and focus efficiency of the microscopic scanning system of the present invention when it is working.

[0059] In a specific embodiment, calibrating the microscopic scanning system includes: S111, placing a calibration sheet 60 on a glass slide carrier 10, and allowing the upper surface of the calibration sheet 60 to be adsorbed and positioned by a vacuum adsorption device 20; wherein the calibration sheet 60 includes a second identification portion 61; controlling the movement of the glass slide carrier 10, and allowing the microscopic camera system 40 to be located directly above the second identification portion 61; controlling the microscopic camera system 40 to move in a height direction, and taking a plurality of images during the movement of the microscopic camera system 40; obtaining a focus reference coordinate Z0 according to the Z-axis coordinate of the image with the largest clarity among the plurality of images obtained during the movement of the microscopic camera system 40.

[0060] The image clarity evaluation algorithm used in calibrating the microscopic scanning system can be implemented by using the evaluation method in the prior art, which will not be described in detail in this disclosure.

[0061] S120, placing the glass slide 50 on the glass slide carrier 10, and allowing the upper surface of the glass slide 50 to be adsorbed and positioned by the vacuum adsorption device 20; wherein the glass slide 50 includes a plurality of first identification portions 51, and an invisible light cutoff coating is provided at the first identification portions 51.

[0062] When the microscopic scanning system of the present invention is used, the structure of the slide 50 can be as follows: Figure 5 As shown, the first identification parts 51 of the glass slide 50 are arranged in multiple rows and columns, and the row spacing and column spacing of the first identification parts 51 are the same. Therefore, when a sample is arranged on the glass slide 50 of the present disclosure, there can be multiple first identification parts 51 around the sample. In a preferred embodiment, the shape of the first identification part 51 is not limited to a circular, rectangular, cross-shaped, or other shapes that are conducive to the clarity evaluation algorithm.

[0063] Correspondingly, after the glass slide 50 is adsorbed and positioned, the upper surface of the glass slide 50 is located at the position of the above-mentioned focus reference coordinate Z0.

[0064] S130 , obtaining N sampling points of the sample according to the position of the sample on the slide 50 .

[0065] In the present disclosure, N sampling points of the sample can be defined according to the position of the sample on the glass slide 50 and the shape of the sample, wherein N is an integer greater than or equal to 1. For example, the sampling points of the sample can be user-defined areas on the sample that require special attention, or can be sampling points evenly distributed on the sample, and the microscopic morphology of the glass slide 50 can be fitted through these sampling points.

[0066] S140, controlling the movement of the slide carrier 10 and making the microscope camera system 40 located at the focus reference coordinates directly above the i-th sampling point; controlling the microscope camera system 40 to move in the height direction, and during the movement of the microscope camera system 40, taking a plurality of non-visible light images including the first identification portion 51; wherein i is greater than or equal to 1 and less than or equal to N.

[0067] Specifically, a low-magnification navigation camera can be used to take a picture of the sample and thereby obtain the position of the sampling point of the sample. The motion mechanism then controls the slide carrier 10 to move to just above the sampling point based on the position of the sampling point obtained by the low-magnification navigation camera.

[0068] Then, the invisible light channel of the light source 30 can be turned on, and the light source 30 will emit infrared rays. At this time, since the first identification part 51 with the invisible light cutoff function is plated on the upper surface of the slide 50, the microscopic camera system 40 is in the focus reference coordinate under the invisible light environment. It can take pictures anywhere and obtain clear black and white mark point images without being affected by samples, oil stains, or impurities.

[0069] S150 , obtaining the Z-axis coordinate Mi corresponding to the non-visible light image with the highest definition including the first identification portion 51 according to the definition of the plurality of non-visible light images taken by the microscope camera system 40 .

[0070] In the present disclosure, the Z-axis coordinate Mi of the first identification portion 51 near the i-th sampling point of the sample is obtained through the clarity evaluation algorithm. In one embodiment, there may be multiple first identification portions 51 near the i-th sampling point, so the Z-axis coordinate Mi of the first identification portion 51 may also be multiple.

[0071] The distribution and density of the first identification portion 51 disclosed in the present invention are adapted to the magnification of the microscope camera system, that is, it is ensured that the uniformly distributed first identification portion 51 can be collected directly below the sample; in a visible light environment, the first identification portion 51 that cuts off invisible light will not affect the shooting and collection of the sample, thereby solving the problem that the first identification portion 51 and the sample area cannot overlap in the Z-axis direction and there cannot be oil stains or impurity interference above the marking point, and the selected first identification portion 51 is directly below the sample, and the fitted microscopic morphology of the slide 50 is more realistic and accurate.

[0072] S160 , obtaining a lower scanning reference plane according to a plurality of Z-axis coordinates Mi corresponding to a plurality of sampling points.

[0073] According to the obtained Z-axis coordinate of the first identification part 51, the microscopic morphology of the upper surface of the slide glass 50 can be fitted, and the upper surface of the slide glass 50 is the lower scanning reference surface. The fitting algorithm is not limited to the triangular patch method, the spline interpolation method, etc.

[0074] S170, performing multi-layer scanning above the lower scanning reference plane, obtaining multiple intermediate images, and fusing the intermediate images to obtain a scanned image.

[0075] Since the sample is at a fixed height above the glass slide 50, according to the acquired lower scanning reference plane (reference focal curved surface), it is possible to +h±k (where h is the theoretical height difference between the sample and the glass slide 50, and k is the deviation value) position, and at this time, the visible light channel of the light source 30 is turned on. Since the invisible light cutoff coating only cuts off the invisible light and does not affect the sample imaging, multi-layer scanning can be performed and image fusion can be performed to obtain a scanned image.

[0076] In addition, the microscopic scanning method disclosed in the present invention can also perform rapid microscopic scanning on a sample including a cover glass 70; wherein a cover glass 70 is provided on the slide glass 50, and the sample is located between the slide glass 50 and the cover glass 70, wherein a plurality of third identification portions 71 are provided on the cover glass 70, and an invisible light cutoff coating is provided at the third identification portion 71.

[0077] In the present disclosure, the first identification portion 51 is formed on the upper surface of the slide glass 50, and the third identification portion 71 is formed on the lower surface of the cover glass 70. In order to facilitate the definition algorithm to distinguish different marking points, the third identification portion 71 of the cover glass 70 has a different shape from the first identification portion 51 of the slide glass 50, and the shape of the third identification portion 71 is not limited to a circular, rectangular, cross, or other shapes that are conducive to the definition evaluation algorithm.

[0078] Similarly, when taking photos in an invisible light environment, the microscope camera system 40 (including a partial invisible light collection function) can capture a clear image of the third identification portion 71 without being affected by the sample, oil stains, and impurities, and the distribution and density of the third identification portion 71 are adapted to the magnification of the microscope camera system, that is, it is ensured that the evenly distributed third identification portion 71 can be collected directly above the sample; in a visible light environment, the third identification portion 71 where the invisible light is cut off will not affect the shooting and collection of the sample.

[0079] The microscopic scanning method disclosed in the present invention may further include: obtaining an upper scanning reference plane through the third identification portion 71 of the cover glass 70; and performing multi-layer scanning at a position between the lower scanning reference plane and the upper scanning reference plane when the microscopic scanning system is performing a scanning operation.

[0080] Specifically, obtaining the upper scanning reference plane through the third identification portion 71 of the cover glass 70 may include: controlling the movement of the slide carrier 10 and making the microscope camera system 40 located at the focus reference coordinates directly above the i-th sampling point; controlling the microscope camera system 40 to move in the height direction, and during the movement of the microscope camera system 40, taking multiple non-visible light images including the third identification portion 71; wherein i is greater than or equal to 1 and less than or equal to N; obtaining the Z-axis coordinate Mi corresponding to the non-visible light image including the third identification portion 71 with the greatest clarity according to the clarity of the multiple non-visible light images taken by the microscope camera system 40; and obtaining the upper scanning reference plane according to the multiple Z-axis coordinates Mi corresponding to the multiple sampling points.

[0081] Since the sample is fixed in the middle of the glass slide 50 and the cover glass 70, the lower scanning reference plane F1 and the upper scanning reference plane F2 can be obtained. Multi-layer scanning is performed at the position of F1+h1±k1 to F2-h2±k2 (wherein h1 is the theoretical height difference between the sample and the slide 50, h2 is the theoretical height difference between the sample and the cover glass 70, and k1 and k2 are deviation values). At this time, the visible light channel of the light source 30 is turned on. Since the invisible light cutoff coating only cuts off the invisible light and does not affect the imaging of the sample, multi-layer scanning can be performed and image fusion can be performed to obtain a scanned image.

[0082] The microscopic scanning method disclosed in the present invention solves the problems that the marking points (the first marking part 51 and the third marking part 71) cannot overlap with the sample area in the Z-axis direction and there cannot be oil stains or impurities above the marking points by taking pictures with an invisible light cutoff coating in conjunction with a microscopic camera system 40 (a visible light camera with a partial invisible light collection function). The selected marking points are located directly below / above the sample, and the fitted microscopic morphology of the glass slide 50 / cover glass 70 is more realistic and accurate. The focus reference point of the glass slide 50 of each device is determined by vacuum adsorption of the calibration piece 60, which greatly reduces the focus range during the autofocus process and realizes rapid fitting of the microscopic morphology of the glass slide 50 / cover glass 70, thereby achieving rapid scanning.

[0083] Specifically: the present invention adopts the invisible light cutoff coating method. Since the marking point has the invisible light cutoff function, the Z-axis coordinates of the marking point can be effectively obtained by taking pictures of the marking point area in an invisible light environment without interfering with the sample, thereby solving the problem that the marking point and the sample cannot overlap in the Z direction, and the selected marking point is just below / above the sample, and the fitted microscopic morphology of the slide 50 / cover glass 70 is more realistic and accurate.

[0084] In addition, the invisible light cutoff coating ensures that the identification of the marking points is not interfered by oil stains and impurities, and there is no need to frequently clean the glass slide 50. The marking point image on the glass slide 50 can be effectively obtained, thereby improving the stability of the measurement.

[0085] When the microscopic scanning system of the present invention leaves the factory, the focus reference point of the glass slide 50 of each device is determined by vacuum adsorption of the special calibration piece 60, which greatly reduces the focus range during the automatic focusing process, and realizes rapid fitting of the microscopic morphology of the glass slide 50, thereby realizing rapid scanning.

[0086] The present disclosure also provides an electronic device, including: a memory, the memory storing execution instructions; and a processor or other hardware module, the processor or other hardware module executes the execution instructions stored in the memory, so that the processor or other hardware module executes the above method.

[0087] The present disclosure also provides a readable storage medium, in which execution instructions are stored. When the execution instructions are executed by a processor, they are used to implement the above method.

[0088] Those skilled in the art should understand that the above embodiments are only for the purpose of clearly illustrating the present disclosure, and are not intended to limit the scope of the present disclosure. For those skilled in the art, other changes or modifications may be made based on the above disclosure, and these changes or modifications are still within the scope of the present disclosure.

Claims

1. A microscopic scanning system, characterized in that: include: Slide carrier; A vacuum adsorption device, the vacuum adsorption device is arranged on the glass slide carrier, so as to adsorb the upper surface of the calibration sheet through the vacuum adsorption device, and make the distance between the upper surface of the calibration sheet and the glass slide carrier fixed; A light source, located below the slide carrier, for providing light; as well as The microscope camera system is located above the slide carrier. The light emitted by the light source passes through the calibration film and then irradiates the microscope camera system to obtain an image of the calibration film, and the focus reference coordinates are obtained based on the image of the calibration film.

2. The microscopic scanning system according to claim 1, characterized in that: The vacuum adsorption device includes at least three vacuum suction cup assemblies; a through hole is opened on the glass slide carrier, and at least three vacuum suction cup assemblies are respectively located at the corners of the through hole of the glass slide carrier, and the calibration piece is adsorbed by the vacuum suction cup assembly so that the upper surface of the calibration piece is fixed relative to the glass slide carrier.

3. The microscopic scanning system according to claim 2, characterized in that: The vacuum suction cup assembly comprises: A base, wherein a gas circulation channel is provided at the upper end of the base; a receiving space is formed at the lower end of the base, the upper surface of the receiving space is formed as a positioning surface, and positioning beads are arranged in the receiving space; an adsorption hole is also provided at the lower end of the base, the adsorption hole is located at the periphery of the receiving space, and the adsorption hole is connected with the gas circulation channel; when the calibration piece is adsorbed by the adsorption hole of the vacuum suction cup assembly, the positioning beads are in contact with the upper surface of the calibration piece and the positioning surface respectively.

4. The microscopic scanning system according to claim 3, characterized in that: The vacuum suction cup assembly also includes: The elastic component is disposed on the base, and the lower end of the elastic component extends out of the lower end of the base.

5. A microscopic scanning method, characterized in that: include: Obtaining the focus reference coordinates of the microscanning system; Placing a glass slide on a glass slide carrier, and allowing the upper surface of the glass slide to be adsorbed and positioned by a vacuum adsorption device; wherein a plurality of first marking portions are provided on the upper surface of the glass slide, and an invisible light cutoff coating film is provided at the first marking portions; Obtaining N sampling points of the sample according to the position of the sample on the slide, wherein N is an integer greater than or equal to 1; Control the movement of the slide carrier and make the microscope camera system located at the focus reference coordinate directly above the i-th sampling point; control the microscope camera system to move in the height direction, and during the movement of the microscope camera system, take a plurality of non-visible light images including the first identification portion; wherein i is greater than or equal to 1 and less than or equal to N; According to the clarity of the multiple non-visible light images taken by the microscope camera system, a Z-axis coordinate Mi corresponding to the non-visible light image with the largest clarity including the first identification portion is obtained; According to a plurality of Z-axis coordinates Mi corresponding to a plurality of sampling points, a lower scanning reference plane is obtained; A multi-layer scan is performed above the lower scanning reference plane to obtain a plurality of intermediate images, which are then fused to obtain a scanned image.

6. The microscopic scanning method according to claim 5, characterized in that: The focusing reference coordinates of the microscopic scanning system are obtained by reading the parameters of the microscopic scanning system; wherein the parameters of the microscopic scanning system are obtained by calibrating the microscopic scanning system.

7. The microscopic scanning method according to claim 6, characterized in that: Calibration of the microscopy scanning system includes: Placing the calibration sheet on the slide carrier, and allowing the upper surface of the calibration sheet to be adsorbed and positioned by the vacuum adsorption device; wherein the calibration sheet includes a second identification portion; Controlling the movement of the slide carrier so that the microscope camera system is located directly above the second identification portion; controlling the microscope camera system to move in a height direction, and capturing a plurality of images including the second identification portion during the movement of the microscope camera system; The focus reference coordinates are obtained according to the Z-axis coordinate of the image with the highest definition among the multiple images obtained by the microscope camera system during the movement.

8. The microscopic scanning method according to claim 6, characterized in that: A cover glass is arranged on the glass slide, and the sample is located between the glass slide and the cover glass, wherein a plurality of third identification parts are arranged on the lower surface of the cover glass, and an invisible light cut-off coating film is arranged at the third identification parts; the microscopic scanning method comprises: The upper scanning reference plane is obtained through the third identification part of the cover glass; when the microscopic scanning system is performing a scanning operation, multi-layer scanning is performed at a position between the lower scanning reference plane and the upper scanning reference plane.

9. The microscopic scanning method according to claim 8, characterized in that: The third identification portion of the cover glass has a shape different from that of the first identification portion of the slide glass.

10. The microscopic scanning method according to claim 5, characterized in that: When the glass slide is positioned by the vacuum adsorption device, the upper surface of the glass slide is located at the focus reference coordinates.

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