A sample liquid containing device and a three-dimensional microscopic imaging system

Abstract

This invention discloses a sample liquid containing device and a three-dimensional microscopic imaging system, relating to the field of medical microscopic detection technology. The sample liquid containing device includes: a glass substrate and a coverslip; the glass substrate is a cylinder with a through-groove extending along its length and penetrating both bottom surfaces of the cylinder, the through-groove occupying one edge of the cylinder and penetrating two adjacent side surfaces; the coverslip covers the two side surfaces of the through-groove exposed along the length of the glass substrate; the space enclosed by the through-groove and the coverslip constitutes a sample liquid containing groove; at least one end of the coverslip along its length is offset from the end of the glass substrate by a predetermined distance to form a sample liquid aspiration port; the size of the sample liquid containing groove is set to allow the liquid-based cells to be detected to pass through and to generate sufficient capillary force to aspirate the sample liquid. This invention enables the microscopic imaging device to capture images of liquid-based cells from different angles and then synthesize them into a realistic three-dimensional microscopic image.

Description

Technical Field

[0001] This invention relates to the field of medical microscopic detection technology, and in particular to a sample liquid containing device and a three-dimensional microscopic imaging system. Background Technology

[0002] With the development of image processing and digital imaging technologies, digital microscopy can be used to scan and image liquid-based cells, obtaining digital images. This allows pathologists or doctors to remotely review slides based on digital images, thereby making accurate diagnoses for difficult cases. Two-dimensional microscopy is gradually becoming insufficient to meet the requirements of medical practice. To increase the information content of cell images and improve the diagnostic accuracy of doctors, three-dimensional imaging of liquid-based cell samples is often necessary.

[0003] Existing 3D imaging technologies often utilize microscope cameras to image a traditional slide and coverslip in three dimensions (X, Y, and Z). Images at different focal lengths are captured along the depth direction (Z-axis) and then synthesized into a 3D image. However, the resulting 3D image is a pseudo-3D image because it only provides 3D information from a single angle and at different depths, failing to provide 3D information from different perspectives and depths. This pseudo-3D image has limitations in identifying certain specific liquid-based cells, such as cancer cells. The pseudo-3D image cannot identify the lateral angle characteristics of the liquid-based cells, such as lateral defects and overlaps, which is crucial for determining whether a sample of liquid-based cells is benign or malignant.

[0004] Because traditional slide and cover glass devices can only take pictures at a vertical angle and thus obtain two-dimensional image data at a vertical angle, they cannot achieve three-dimensional imaging from multiple angles. Therefore, existing technologies based on traditional slide devices cannot achieve true three-dimensional microscopic imaging of liquid-based cells. Summary of the Invention

[0005] In view of the above-mentioned defects or deficiencies in the prior art, the present invention provides a sample liquid containing device and a three-dimensional microscopic imaging system, which completely abandons the traditional thin sheet structure composed of a glass slide and a coverslip, and creatively adopts a three-dimensional structure composed of a glass slide and a coverslip. This allows the microscopic imaging device to capture liquid-based cell images from different angles and then synthesize them into a real three-dimensional microscopic image. Doctors can obtain multi-angle three-dimensional information of liquid-based cells through this three-dimensional microscopic image, providing strong support for doctors' diagnostic decisions.

[0006] One aspect of the present invention provides a sample liquid containing device, comprising:

[0007] Glass carrier and coverslip;

[0008] The glass carrier is a column with a through groove. The column includes two adjacent perpendicular sides. The through groove extends along the length of the column and penetrates the two bottom surfaces of the column. The through groove occupies a right-angled side edge of the column and penetrates the two adjacent perpendicular sides of the column.

[0009] The coverslip is fixed to the glass carrier and covers two adjacent vertical sides of the through groove in the length direction of the glass carrier; the space enclosed by the through groove and the coverslip constitutes a sample liquid receiving groove; at least one end of the coverslip in the length direction is offset from the bottom end of the glass carrier by a predetermined distance to form a sample liquid suction port of the sample liquid receiving groove.

[0010] The sample solution container is sized to allow the liquid-based cells to be tested to pass through and to generate sufficient capillary force to draw in the sample solution.

[0011] Furthermore, the sample liquid container has a square cross-section with a side length of 20-30 μm.

[0012] Furthermore, the column is a cuboid with a square base, the length of which is 5-10cm and the side length of the square base is 0.5-0.7cm.

[0013] Furthermore, the cover glass is detachably connected to the glass carrier.

[0014] Furthermore, the cover glass is snapped together with the glass carrier.

[0015] Furthermore, the two adjacent vertical sides of the glass body penetrated by the through groove are each covered with an independent cover glass, and at the intersection of the two independent cover glass, one cover glass completely covers the side of the other cover glass.

[0016] Another aspect of the present invention provides a three-dimensional microscopic imaging system, comprising:

[0017] The aforementioned sample liquid containing device, microscope imaging device, light source, and controller; wherein,

[0018] The microscopic imaging device includes a first camera and a second camera aligned with two adjacent vertical sides penetrated by the through slot. The first camera and the second camera are located on the same plane and are perpendicular to each other.

[0019] The light source is used to illuminate the sample liquid container through the glass carrier;

[0020] The controller is used to control the first camera and the second camera to move from one end of the sample liquid container to the other end at a predetermined shooting frequency and moving speed, and to send the acquired image data to the host computer.

[0021] Furthermore, the glass carrier is a cuboid; the light source includes a first light source and a second light source, the first light source being perpendicularly aligned with a first side of the glass carrier that does not expose the through groove, and the second light source being perpendicularly aligned with a second side of the glass carrier that does not expose the through groove.

[0022] Furthermore, the glass carrier is a cuboid; the light source illuminates the glass carrier along the opposite edges of the through groove on the glass carrier.

[0023] Furthermore, the edges opposite to the through groove of the glass carrier are cut off, with the cut surface perpendicular to the incident light from the light source.

[0024] The present invention provides a sample liquid containing device and a three-dimensional microscopic imaging system, which have the following features:

[0025] Beneficial effects:

[0026] (1) The traditional planar structure of the glass slide is replaced by the three-dimensional structure of the glass body, which enables the imaging device to capture images of liquid-based cells from different angles, thus facilitating the synthesis of real three-dimensional microscopic images;

[0027] (2) The structure of the sample liquid container on the glass body ensures that the microscope imaging device is not affected by the uneven surface of the etched groove or laser-cut groove when taking pictures, and can obtain clearer image data.

[0028] (3) The glass slide, coverslip and through groove form a capillary structure sample liquid receiving groove, which can be automatically filled by capillary force when injecting sample liquid, which is very convenient.

[0029] (4) By using two orthogonal microscopic cameras to take multi-angle pictures of the sample liquid container at a predetermined shooting frequency and moving speed, and then performing three-dimensional synthesis of the pictures, a real three-dimensional microscopic image with a much greater amount of information than traditional technology can be obtained. Attached Figure Description

[0030] Other features, objects, and advantages of this application will become more apparent from the following detailed description of non-limiting embodiments with reference to the accompanying drawings:

[0031] Figure 1 This is a schematic diagram of a thin-film sample liquid containing a glass slide and a coverslip in the prior art;

[0032] Figure 2 This is a cross-section of a sample liquid containing device provided in one embodiment of this application. Figure 1 ;

[0033] Figure 3 This is a cross-section of a sample liquid containing device provided in another embodiment of this application. Figure 2 ;

[0034] Figure 4 This is a perspective view of a sample liquid containing device provided in another embodiment of this application;

[0035] Figure 5 This is a schematic diagram of a three-dimensional microscopic imaging system provided in another embodiment of this application. Figure 1 ;

[0036] Figure 6 This is a schematic diagram of a three-dimensional microscopic imaging system provided in another embodiment of this application. Figure 2 .

[0037] Figure label:

[0038] 100-Thin-film sample liquid receiving device; 101-Slide; 102-Covering slip; 103-Microscope objective lens; 104-Sample liquid; 200-Three-dimensional sample liquid receiving device; 201-Slide body; 202-Covering slip; 203-Through groove; 204-Sample liquid receiving groove; 205-Sample liquid suction port; 206-Card slot; 207-Snap fastener; 300-Three-dimensional microscopic imaging system; 301-First camera; 302-Second camera; 303-First light source; 304-Second light source; 305-First side; 306-Second side. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0040] The terminology used in the embodiments of this invention is for the purpose of describing particular embodiments only and is not intended to limit the invention. The singular forms “a,” “the,” and “the” as used in the embodiments of this invention and the appended claims are also intended to include the plural forms unless the context clearly indicates otherwise.

[0041] It should be understood that although the terms first, second, third, etc., may be used to describe the acquisition modules in the embodiments of the present invention, these acquisition modules should not be limited to these terms. These terms are only used to distinguish the acquisition modules from each other.

[0042] Depending on the context, the word "if" as used here can be interpreted as "when," "when," "in response to determination," or "in response to detection." Similarly, depending on the context, the phrase "if determination" or "if detection (of the stated condition or event)" can be interpreted as "when determination," "in response to determination," "when detection (of the stated condition or event)," or "in response to detection (of the stated condition or event)."

[0043] It should be noted that the directional terms such as "upper," "lower," "left," and "right" used in the embodiments of the present invention are used to describe the angles shown in the accompanying drawings and should not be construed as limiting the embodiments of the present invention. Furthermore, in the context, it should be understood that when it is mentioned that an element is formed "upper" or "lower" of another element, it can not only be formed directly "upper" or "lower" of the other element, but also indirectly "upper" or "lower" of the other element through an intermediate element.

[0044] Figure 1 This illustration shows a sheet-type sample liquid containing device 100, consisting of a glass slide 101 and a coverslip 102, as used in the prior art. The glass slide 101 refers to the object-carrying glass slide, and the coverslip 102 refers to the covering glass slide. Typically, the coverslip 102 is positioned on the side of the glass slide 101 closest to the microscope objective lens 103, with the sample liquid 104 placed between the glass slide 101 and the coverslip 102. This conventional sheet-type sample liquid containing device 100 only allows the microscope lens or microscopic imaging device to observe or photograph vertically downwards from above the coverslip 102. While it can photograph samples at different focal depths, it cannot acquire images of liquid-based cells in the sample liquid from other angles. Therefore, because the sheet-type sample liquid containing device 100 cannot acquire multi-angle images of liquid-based cells in the sample liquid, it is impossible to synthesize a true three-dimensional image containing multi-angle information.

[0045] To overcome the shortcomings of the traditional thin-film sample liquid container 100, see [reference needed]. Figure 2-4 One embodiment of the present invention provides a sample liquid containing device 200 with a three-dimensional structure, including: a glass carrier 201 and a coverslip 202. The glass carrier 201 is a cylindrical structure with a through groove 203. In this embodiment, the cylinder can be a cylinder, a prism, or any other type of cylinder with the same and parallel base shape. The cylinder must also include two adjacent perpendicular sides, such as a cuboid, a cube, a right-angled triangular prism, a right-angled trapezoidal prism, etc. For ease of description, a cuboid will be used as an example below.

[0046] A through groove 203 is formed on a cuboid-shaped glass substrate 201 by etching or laser cutting. Since glass etching and laser cutting are existing technologies in the field, the specific process of forming the through groove 203 will not be described in this embodiment. Further, the through groove 203 extends along the length of the cuboid and penetrates both bottom surfaces of the cuboid. The through groove 203 occupies one edge of the cuboid and penetrates two adjacent side surfaces. A cover glass 202 is fixed to the glass substrate 201 and covers the two exposed side surfaces of the through groove 203 along the length of the glass substrate 201. The space enclosed by the through groove 203 and the cover glass 202 constitutes a sample liquid receiving tank 204. At least one end of the cover glass 202 along its length is offset from the bottom end of the glass substrate 201 by a predetermined distance to form a sample liquid suction port 205 of the sample liquid receiving tank 204. The sample liquid receiving tank 204 is sized to allow the liquid-based cells to be tested to pass through and to generate sufficient capillary force to draw in the sample liquid.

[0047] It should be noted that in this embodiment, the sample liquid receiving groove 204 ultimately occupies one edge of the cuboid structure of the glass substrate 201. That is, the sample liquid receiving groove 204 penetrates the edge and two adjacent side surfaces of the cuboid along its length, making the sample liquid receiving groove 204 open on two adjacent side surfaces of the cuboid. In other words, a right-angled edge of the column (taking a cuboid as an example) is removed to form a through groove 203. The camera can directly take microscopic pictures at different angles through the coverslip 202 covering the two adjacent side surfaces penetrated by the through groove 203, without having to take pictures through the glass substrate 201. The advantages of the sample liquid receiving tank 204 arranged in the above manner are as follows: if etched grooves or laser-cut grooves are formed in other parts, the grooves will either have only one exposed surface on the outer side of the cuboid, or the grooves will be completely closed by the glass substrate 201 with only the bottom surface being open. This means that at most one camera can take pictures through the coverslip 202, while other cameras can only take pictures through the etched or cut surfaces of the grooves on the glass substrate 201. However, the glass surface formed by etching or laser cutting is not smooth, and taking pictures through the etched or cut surfaces will affect the shooting effect of the microscope cameras, thus affecting the final three-dimensional imaging effect. The coverslip 202 is a glass slide made by a special deposition process, with a smooth surface and high light transmittance. The arrangement of the sample liquid receiving tank 204 in this embodiment allows two cameras at different angles to directly capture pictures of the sample liquid receiving tank 204 through the coverslip 202, greatly improving the image quality of the microscope imaging.

[0048] In another embodiment, the length of the cuboid in the sample solution container 200 is set to 5-10 cm, and the cross-section and base of the cuboid are square, with the side length of the square set to 0.5-0.7 cm. Taking cancer cells as an example, liquid-based cells are typically 10-20 μm in size. The sample solution container 204 must be able to accommodate at least the liquid-based cells to be tested; therefore, the width or diameter of the sample solution container 204 should not be less than 20 μm. Simultaneously, the sample solution container 204 needs to utilize capillary force generated by the capillary tube to draw in the dripped sample solution; therefore, the diameter or width of the sample solution container 204 should ensure sufficient capillary force. Considering the above factors, this embodiment, after extensive experimental verification, demonstrates that setting the diameter or width of the sample solution container 204 to 20-30 μm allows most types of liquid-based cells to pass through while generating sufficiently large capillary force.

[0049] Furthermore, the cover glass 202 is fixedly connected to the glass carrier 201, for example, by bonding.

[0050] Further, see Figure 2-3 The coverslip 202 and the glass carrier 201 are connected by a detachable structure, for example, by a snap-fit ​​connection. Specifically, a slot 206 is provided on the side of the glass carrier 201, and a snap 207 matching the slot 206 is provided on the coverslip 202. By aligning the snap 207 of the coverslip 202 with the slot 206 on the glass carrier 201 and pressing it, the coverslip 202 can be pressed tightly against the glass carrier 201. This snap-fit ​​structure makes the assembly and disassembly of the coverslip 202 and the glass carrier 201 more convenient, thereby simplifying the manufacturing process.

[0051] Furthermore, participate Figure 2 The cover glass 202 can be an integral structure bent at a right angle, so that it can be installed as a whole on the glass carrier and cover the two adjacent sides of the cuboid with the through groove 203 exposed.

[0052] Further, see Figure 3 The coverslip 202 can be two independent parts, each covering one of the two adjacent sides of the cuboid that expose the through groove 203. Preferably, at the intersection of the two independent coverslips 202, one coverslip 202 completely covers the side of the other coverslip 202, so that the coverslips form a surface contact, which makes the seal more tight and ensures that the sample liquid in the sample liquid receiving tank 204 will not overflow.

[0053] Further, see Figure 4At least one end of the coverslip 202 is offset from the bottom end of the glass substrate 201 by a predetermined distance in the longitudinal direction to form a sample liquid aspiration port 205. The predetermined offset distance is preferably 2 to 4 mm, which is more conducive to rapid aspiration of sample liquid. A predetermined distance of less than 2 mm makes it difficult to aspirate sample liquid, while a predetermined distance of more than 4 mm wastes the effective space of the sample liquid receiving tank 204.

[0054] The sample liquid containing device provided in this embodiment replaces the traditional thin-film structure with a three-dimensional structure of a glass substrate, which facilitates two imaging devices to capture images of liquid-based cells from different angles, thereby synthesizing a realistic three-dimensional microscopic image. The sample liquid containing groove is located at the corner of the glass substrate, so that the two microscopic imaging devices at different angles are not affected by the uneven surface of the etched grooves or laser-cut grooves when taking pictures, and can obtain clearer image data. The glass substrate, coverslip and through groove form a capillary structure sample liquid containing groove, which can automatically fill the sample liquid containing groove by capillary force when injecting sample liquid, which is very convenient.

[0055] See Figure 5-6 Another embodiment of the present invention provides a three-dimensional microscopic imaging system 300, including the sample liquid containing device 200, microscopic imaging device, light source and controller in the above embodiments, wherein the glass carrier is illustrated by way of a cuboid shape.

[0056] The microscope imaging device includes a first camera 301 and a second camera 302 that are perpendicularly aligned with two cover glass slides 202 covering the through groove 203 (i.e., perpendicularly aligned with two adjacent sides pierced by the through groove). The first camera 301 and the second camera 302 are located on the same plane and are perpendicular to each other.

[0057] A light source is used to illuminate the sample solution container 204 through the glass substrate 201. See also Figure 5 In one optional embodiment, the light source includes a first light source 303 and a second light source 304. The first light source 303 is aligned with a first side 305 of the glass carrier 201 that does not expose the through groove 203, and the second light source 304 is aligned with a second side 306 of the glass carrier 201 that does not expose the through groove 203. See also Figure 6 In another alternative embodiment, the light source illuminates the glass substrate 201 along the opposing edges of the through groove 203 on the glass substrate 201. The light is split in two and enters the glass substrate 201 on both sides of the edge, thereby providing illumination to the sample liquid receiving tank 204. More preferably, the opposing edges of the through groove 203 of the glass substrate 201 are cut off, with the cut surface perpendicular to the incident light from the light source. This allows for the introduction of more light even with only one or a group of light sources.

[0058] Furthermore, the controller controls the first camera 301 and the second camera 302 to move from one end of the sample liquid receiving tank 204 to the other end at a predetermined shooting frequency and moving speed, and sends the acquired image data from the two different angles to the host computer, which then synthesizes them into a three-dimensional image. More preferably, the controller moves the first camera 301 and the second camera 302 from one end of the coverslip 202 to the other end of the coverslip 202 during shooting. Only the area covered by the coverslip 202 is photographed because: firstly, the coverslip 202 is made using a deposition process, so it has high light transmittance and is not affected by the uneven etched surface or laser-cut surface of the through groove 203; secondly, the standard cuboid receiving space formed after the coverslip 202 covers the through groove 203 is filled with sample liquid, and the image quality of the regularly shaped, bubble-free sample liquid is better.

[0059] Optionally, the controller can also be used to control the on / off state of the light source.

[0060] The above description is merely a preferred embodiment of the present invention. Those skilled in the art should understand that the scope of disclosure involved in the present invention is not limited to the technical solutions formed by specific combinations of the above-described technical features, but should also cover other technical solutions formed by arbitrary combinations of the above-described technical features or their equivalents without departing from the above-described concept. For example, technical solutions formed by substituting the above-described features with (but not limited to) technical features with similar functions disclosed in the present invention.

Claims

1. A sample liquid containing device for three-dimensional microscopic imaging, characterized in that, include: Glass carrier and coverslip; The glass carrier is a column with a through groove. The column includes two adjacent perpendicular sides. The through groove extends along the length of the column and penetrates the two bottom surfaces of the column. The through groove occupies a right-angled side edge of the column and penetrates the two adjacent perpendicular sides of the column. The coverslip is fixed to the glass carrier and covers two adjacent vertical sides of the through groove in the length direction of the glass carrier; the space enclosed by the through groove and the coverslip constitutes a sample liquid receiving groove; at least one end of the coverslip in the length direction is offset from the bottom end of the glass carrier by a predetermined distance to form a sample liquid suction port of the sample liquid receiving groove. The sample solution container is sized to allow the liquid-based cells to be tested to pass through and to generate sufficient capillary force to draw in the sample solution.

2. The sample liquid containing device for three-dimensional microscopic imaging according to claim 1, characterized in that, The sample liquid container has a square cross-section with a side length of 20-30 mm.

3. A sample liquid containing device for three-dimensional microscopic imaging according to claim 2, characterized in that, The column is a cuboid with a square base, the length of which is 5-10cm and the side length of the square base is 0.5-0.7cm.

4. A sample liquid containing device for three-dimensional microscopic imaging according to claim 1, characterized in that, The cover glass is detachably connected to the glass carrier.

5. A sample liquid containing device for three-dimensional microscopic imaging according to claim 4, characterized in that, The cover glass is snapped together with the glass carrier.

6. A sample liquid containing device for three-dimensional microscopic imaging according to claim 1, characterized in that, The glass substrate is covered by two adjacent vertical sides through which the through groove passes, and at the intersection of the two independent cover sheets, one cover sheet completely covers the side of the other cover sheet.

7. A three-dimensional microscopic imaging system, characterized in that, include: The sample liquid containing device, microscopic imaging device, light source, and controller for three-dimensional microscopic imaging as described in any one of claims 1-6; wherein, The microscopic imaging device includes a first camera and a second camera aligned with two adjacent vertical sides penetrated by the through slot. The first camera and the second camera are located on the same plane and are perpendicular to each other. The light source is used to illuminate the sample liquid container through the glass carrier; The controller is used to control the first camera and the second camera to move from one end of the sample liquid container to the other end at a predetermined shooting frequency and moving speed, and to send the acquired image data to the host computer.

8. A three-dimensional microscopic imaging system according to claim 7, characterized in that, The glass carrier is a cuboid; The light source includes a first light source and a second light source. The first light source is perpendicularly aligned with a first side of the glass carrier that does not expose the through groove, and the second light source is perpendicularly aligned with a second side of the glass carrier that does not expose the through groove.

9. A three-dimensional microscopic imaging system according to claim 7, characterized in that, The glass carrier is a cuboid; The light source illuminates the glass substrate along the opposite edges of the through groove on the glass substrate.

10. A three-dimensional microscopic imaging system according to claim 9, characterized in that, The edges opposite to the through groove of the glass carrier are cut off, and the cut surface is perpendicular to the incident light from the light source.

Images