Sectional-immersion imaging system and method suitable for high-resolution three-dimensional imaging of large sample

Abstract

The present application discloses a sectional-immersion imaging system suitable for high-resolution three-dimensional imaging of a large sample, comprising: a liquid imaging medium immersion circulation module, an illumination imaging module, and a cutting module. In the present application, by means of the liquid imaging medium immersion circulation module, a stable liquid imaging medium immersion environment can be only formed between a photographing unit and a sample in the process of using a mechanical cutting-based imaging method during three-dimensional imaging of a large biological sample. Unlike existing methods that a sample containing embedded biological tissue is immersed in a liquid imaging medium, the present application can achieve high-resolution imaging by using only a small amount of a liquid imaging medium capable of forming a liquid imaging medium immersion environment, such that the situation that cutting performance is affected by sample deformation due to prolonged immersion of a sample in a high-refractive-index liquid imaging medium is effectively avoided, then the amount of the liquid imaging medium used in a water tank is further reduced, thereby lowering the load requirements for a three-dimensional translation stage while reducing the use cost.

Description

A cross-sectional immersion imaging system and method suitable for high-resolution three-dimensional imaging of large samples Technical Field

[0001] This application relates to the research and medical fields, specifically to a cross-sectional immersion imaging system and method suitable for large-sample, high-resolution three-dimensional imaging. Background Technology

[0002] In biological and medical research, various automated microscopic optical imaging systems developed in recent years can achieve complete three-dimensional imaging of samples such as the whole brain of mice and monkeys. These systems can be broadly classified into two categories of imaging methods: those based on tissue transparency and those based on mechanical cutting.

[0003] One imaging method based on tissue transparency involves making biological tissue transparent to increase the depth of optical imaging, followed by rapid imaging using light-sheet fluorescence microscopy. This method is suitable for large-scale, rapid imaging, but due to the refractive index mismatch remaining in deeper layers of the sample, the imaging resolution and quality decrease significantly with increasing imaging depth.

[0004] Imaging methods based on mechanical cutting combine optical tomography with precision cutting techniques. Biological tissue is embedded in a specific medium to form a sample, achieving the hardness required for precision cutting. During sample imaging, the sample is placed on a three-dimensional translation stage. The objective lens focuses illumination light onto the shallow portion of the sample cross-section and collects imaging light. Driven by the three-dimensional translation stage, the sample acquires a tomographic image of the entire shallow cross-section through the illumination and imaging modules. Then, a cutting tool removes the imaged portion of the sample surface. By continuously repeating the "cross-section imaging-surface cutting" process, three-dimensional imaging of centimeter-sized samples can be achieved. The advantage of this method is that the imaging focal plane remains in the shallow layer of the sample, where light scattering and absorption are very low, ensuring uniform and consistent imaging quality for large samples and enabling the acquisition of high-resolution three-dimensional imaging data.

[0005] In mechanical cutting-based imaging methods, to improve imaging resolution, objectives adapted to high-refractive-index imaging media are often used, such as water immersion lenses, oil immersion lenses, and other objectives adapted to high-refractive-index imaging media. When using these objectives, the sample generally needs to be fixed in a water bath filled with the imaging medium. However, this approach introduces a series of problems, including:

[0006] (1) Long-term immersion of the sample in a water bath filled with water, oil or other high refractive index imaging media will cause the sample to deform and thus affect the cutting performance.

[0007] (2) A large amount of oil or other high-refractive-index imaging medium is required to fill the water tank. Such media are generally expensive, which will greatly increase the cost of use.

[0008] (3) The weight of the water tank increases significantly after it is filled with imaging medium liquid, which increases the load on the three-dimensional translation stage and affects the dynamic positioning accuracy of the three-dimensional translation stage. Alternatively, a more expensive high-load three-dimensional translation stage may be required. Summary of the Invention

[0009] To address the above problems, this application provides the following technical solution:

[0010] A cross-sectional immersion imaging system suitable for large-sample, high-resolution three-dimensional imaging includes: an imaging medium liquid immersion circulation module, an illumination imaging module, and a cutting module.

[0011] The imaging medium liquid immersion circulation module includes a reservoir configured to store the imaging medium liquid, an immersion assembly connected to the reservoir via a pipe, a medium liquid recovery structure connected to the immersion assembly and the reservoir via a pipe, and a water pump. The imaging medium liquid can complete a full circulation between the reservoir, the medium liquid recovery structure, and the immersion assembly, and the water pump can provide power for this circulation.

[0012] The illumination imaging module is located directly above the sample wetting component and can take pictures of the sample after it has been wetting with the imaging medium. When the imaging medium flows through the wetting component to the sample surface, it can form a stable imaging medium wetting environment between the illumination imaging module and the sample.

[0013] Based on the above technical solution, the following improvements can be made to this application.

[0014] Furthermore, the illumination imaging module includes an objective lens positioned above the sample immersion assembly to focus the illumination light and collect the sample light signal, as well as an illumination and imaging optical path capable of providing stable illumination and imaging the sample light signal collected by the objective lens.

[0015] Furthermore, the cutting module includes a cutting structure capable of cutting the sample surface, a three-dimensional translation stage capable of moving the entire sample to the cutting structure, and a debris collection structure capable of collecting the sample fragments cut off by the cutting structure.

[0016] Furthermore, the immersion assembly includes an immersion member, a groove within the immersion member for the inflow of imaging medium liquid, and an insertion port within the groove that is adapted to the objective lens. After the objective lens enters the insertion port, a gap is left between it and the inside of the insertion port for the imaging medium liquid to flow down.

[0017] Furthermore, the medium liquid recovery structure includes a water tank capable of holding the sample and a filter structure located between the water tank and the reservoir. The filter structure is connected to the water tank and the reservoir via pipes. The pipe structure in the imaging medium liquid immersion circulation module is equipped with a control valve that can control its opening and closing. When the imaging medium liquid in the water tank enters the reservoir, the filter structure can filter the sample debris present in the imaging medium liquid. The water tank is located on the surface of the movable end of the three-dimensional translation stage, and the water tank can be driven by the three-dimensional translation stage to perform three-dimensional motion. The water tank is equipped with a limiting structure that can limit the position of the sample in the water tank.

[0018] Furthermore, the cutting module includes a scraping unit capable of scraping off residual imaging medium liquid on the sample surface before the sample is cut. The scraping unit is located between the objective lens and the cutting structure. The debris collection structure includes a suction head located at the cutting edge of the cutting structure, a debris filter device connected to the suction head via a pipe, and a negative pressure fan that provides suction to the suction head. Both the suction head and the pipe connecting the debris filter device are made of transparent material that can withstand negative pressure.

[0019] Furthermore, the inner groove of the immersion member is provided with a connector for the imaging medium liquid in the reservoir to flow in. A baffle plate is provided at the position between the connector and the socket in the groove to limit the flow rate of the liquid flowing out of the connector. An opening is provided on the outside of the groove for the imaging medium liquid to flow into the water tank. A liquid blocking ring is provided in the groove near the socket, and the surface of the liquid blocking ring is provided with an opening for the imaging medium liquid to flow into the socket.

[0020] A cross-sectional immersion imaging method suitable for large-sample, high-resolution three-dimensional imaging, including the aforementioned cross-sectional immersion imaging system suitable for large-sample, high-resolution three-dimensional imaging, comprising the following steps.

[0021] S1. Fix the sample containing the embedded biological tissue in the water tank and pour the imaging medium into the reservoir;

[0022] S2. The water tank is moved by the three-dimensional translation stage, so that the sample is moved directly below the objective lens. The control valve and water pump in the imaging medium liquid immersion circulation module are opened, so that the imaging medium liquid in the reservoir circulates between the immersion component and the reservoir. At this time, the imaging medium liquid forms a stable imaging medium immersion environment between the objective lens and the sample.

[0023] S3. The water tank is moved by a three-dimensional translation stage, and the illumination imaging module realizes the imaging and recording of the entire sample surface in the imaging medium immersion environment.

[0024] S4. After recording is completed, the three-dimensional translation stage moves the entire water tank. At this time, the sample leaves the bottom of the objective lens and comes into contact with the cutting edge of the cutting structure. The cutting structure removes the sample surface that has been recorded by the illumination imaging module, exposing a new sample cross-section.

[0025] S5. The three-dimensional translation stage moves the sample back to the bottom of the objective lens, and steps S2 to S4 are repeated.

[0026] Furthermore, in step S2, before the imaging medium liquid in the water tank enters the reservoir, the filtration structure filters sample debris in the imaging medium liquid.

[0027] Furthermore, in step S4, after the illumination imaging module records the imaging results of the sample surface and before the three-dimensional translation stage moves the water tank as a whole, the control valves and water pumps in the pipeline of the imaging medium liquid immersion circulation module are closed.

[0028] Furthermore, in step S5, after the illumination imaging module records the imaging results of the sample surface, before the three-dimensional translation stage moves the water tank and the sample as a whole to be cut by the cutting structure, the negative pressure fan in the debris collection structure is started, causing the debris collection structure to collect the cut sample debris.

[0029] Furthermore, in step S4, after the cutting structure has finished cutting the sample, the negative pressure fan is turned off.

[0030] Furthermore, in step S4, before the portion of the sample with the recorded image is cut off by the cutting structure, the scraping unit can scrape off the residual imaging medium liquid on the sample into the water tank.

[0031] Furthermore, the three-dimensional translation stage can raise the overall height of the water tank after each cutting of the sample surface by the cutting structure, so as to prepare for the next cutting of the sample surface.

[0032] Details of one or more embodiments of this application are set forth in the following drawings and description. Other features, objects, and advantages of this application will become apparent from the specification, drawings, and claims. Attached Figure Description

[0033] To better describe and illustrate embodiments and / or examples of the inventions disclosed herein, reference may be made to one or more accompanying drawings. The appendices or examples used to describe the drawings should not be considered as limiting the scope of any of the disclosed inventions, the embodiments and / or examples currently described, or the best mode of these inventions as currently understood.

[0034] Figure 1 is a schematic diagram of a cross-sectional immersion imaging system for large-sample high-resolution three-dimensional imaging according to one or more embodiments;

[0035] Figure 2 is a perspective view of an immersion assembly according to one or more embodiments;

[0036] Figure 3 is a schematic cross-sectional view of the immersion assembly according to one or more embodiments;

[0037] Figure 4 is a schematic diagram of the cross-sectional structure of a cutting module cutting a sample according to one or more embodiments;

[0038] Figure 5 is a cross-sectional structural diagram of a suction head according to one or more embodiments;

[0039] The components represented by each number in the attached diagram are listed below: 1. Imaging medium immersion circulation module; 11. Reservoir; 12. Immersion assembly; 121. Immersion part; 122. Groove; 123. Inlet; 124. Isolation plate; 13. Medium recovery structure; 131. Water tank; 132. Filter structure; 2. Illumination and imaging module; 21. Objective lens; 22. Illumination and imaging optical path; 3. Cutting module; 31. Cutting structure; 32. Three-dimensional translation stage; 33. Debris collection structure; 331. Suction head; 332. Debris filtration device; 333. Negative pressure fan; 34. Scraper unit. Detailed Implementation

[0040] Please refer to Figure 1. This application discloses a cross-sectional immersion imaging system suitable for high-resolution three-dimensional imaging of large samples, including an imaging medium liquid immersion circulation module 1, an illumination imaging module 2 capable of photographing the sample surface, and a cutting module 3 capable of cutting the sample surface.

[0041] The imaging medium immersion circulation module 1 includes a reservoir 11 configured to store the imaging medium, an immersion component 12 connected to the reservoir 11 via a pipe, a medium recovery structure 13 connected to the immersion component 12 and the reservoir 11 via a pipe, and a water pump. The imaging medium can complete a full circulation between the reservoir 11, the medium recovery structure 13, and the immersion component 12. The water pump provides power for this circulation. The pipes in the imaging medium immersion circulation module 1 are equipped with control valves that can control the opening and closing of the pipes. To ensure smooth circulation between the immersion component 12, the reservoir 11, and the water pump, the three can be interconnected via pipes. Through the uninterrupted circulation of the imaging medium, a stable flow of imaging medium can be provided for sample imaging even when the sample is not immersed in the imaging medium.

[0042] The imaging medium can be selected from liquids such as water, aqueous solutions containing dyes, glycerol, immersion oil, and other high-refractive-index imaging mediums with composite components, depending on the imaging requirements.

[0043] The illumination imaging module 2 is an optical and electronic device commonly used in mechanical cutting imaging methods in the prior art. It includes an objective lens 21 located above the sample wetting component 12 to focus the illumination light and collect the sample light signal, and an illumination and imaging optical path 22 that can provide stable illumination and image the sample light signal collected by the objective lens. When the imaging medium liquid flows through the wetting component 12 to the sample surface, it can form a stable imaging medium liquid wetting environment between the objective lens 21 and the sample.

[0044] The imaging medium wetting environment here refers to the liquid environment formed between the imaging medium liquid and the objective lens 21 at the location where the sample needs to be photographed, due to its own surface tension and under stable flow conditions. The imaging medium liquid can form a stable imaging medium liquid wetting environment by controlling the distance between the port 123 and the objective lens 21, the distance between the lower surface of the wetting component 12 and the sample surface, and the microstructure of the lower surface of the wetting component 12.

[0045] The cutting module 3 includes a cutting structure 31 capable of cutting the sample surface, a three-dimensional translation stage 32 capable of moving the entire sample to the cutting structure 31, and a debris collection structure 33 capable of collecting the sample fragments cut off by the cutting structure 31. The three-dimensional translation stage 32 is a common electrical device in the prior art. It can drive the water tank 131 on the three-dimensional translation stage 32 to move horizontally, vertically, and longitudinally through the cooperation of a motor and mechanical structure. During the sample cutting process, the three-dimensional translation stage 32 can adjust the height of the water tank 131 according to the height reduction of the sample after cutting, ensuring that the cutting structure 31 can cut the part of the sample that was not photographed.

[0046] The three-dimensional translation stage 32 can image a large area of ​​sample surface. Since the imaging field of view of the objective lens 21 is relatively small, for a large area of ​​sample surface, the three-dimensional translation stage 32 needs to move the sample to expand the imaging range through scanning imaging.

[0047] Compared with existing technologies, the above technical solution only involves the upper section of the sample in contact with the imaging medium liquid, avoiding problems such as deformation and reduced cutting performance caused by the sample being immersed in the imaging medium liquid for a long time. At the same time, the recyclable imaging medium liquid and the imaging medium liquid environment formed only on the section greatly reduce the amount of imaging medium liquid used in the imaging process of large-volume samples, effectively reducing the cost of use.

[0048] At the same time, the reduction in the amount of imaging medium liquid also greatly reduces the load capacity requirements of the three-dimensional translation stage 32, thereby reducing the system hardware cost.

[0049] To provide a detailed explanation of the above technical solutions, the following embodiments are listed. The following descriptions are merely specific implementations of this application, but the scope of protection of this application is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in this application, based on the technical solutions and inventive concepts of this application, should be covered within the scope of protection of this application.

[0050] Example 1:

[0051] Based on the above technical solution, the immersion assembly 12 includes an immersion member 121, a groove 122 provided in the immersion member 121 for the inflow of imaging medium liquid, and an insertion port 123 provided in the groove 122 that is adapted to the objective lens 21. After the objective lens 21 enters the insertion port 123, a gap is left between it and the inside of the insertion port 123 for the imaging medium liquid to flow down. To facilitate the production and installation of the immersion member 121, the immersion member 121 can be a ring structure as shown in the figure. The immersion member 121 can be fitted onto the surface of the objective lens 21. When the sample is photographed, the imaging medium liquid flowing into the groove 122 inside the immersion member 121 can flow down from the gap between the objective lens and the immersion member 121, forming a stable imaging medium liquid immersion environment between the objective lens 21 and the sample.

[0052] Based on the above, in order to collect and reuse the imaging medium liquid flowing out of the immersion component 121, the medium liquid recovery structure 13 includes a water tank 131 capable of holding the sample and a filter structure 132 disposed between the water tank 131 and the reservoir 11. The filter structure 132 is connected to the water tank 131 and the reservoir 11 through a pipe structure. When the imaging medium liquid in the water tank 131 enters the reservoir 11, the filter structure 132 can filter the sample debris present in the imaging medium liquid. The filter structure 132 can be a common structure composed of a device shell and internal filter material. It is designed to filter out the sample debris contained in the imaging medium liquid entering the reservoir 11 from the water tank 131, ensuring the cleanliness of the imaging medium liquid stored in the reservoir 11 and avoiding affecting the imaging effect of the subsequent illumination imaging module 2.

[0053] Preferably, the bottom of the water tank 131 is inclined, and the bottom of the inclined surface is connected to the pipe leading to the filter structure 132. With the design of the bottom of the water tank 131 being inclined, after the imaging medium liquid flows from the sample surface into the water tank 131, under the action of gravity, the imaging medium liquid can better flow back to the pipe opening and be collected by the water pump into the reservoir 11, so as to reduce the imaging medium liquid accumulated in the water tank 131.

[0054] Example 2:

[0055] Unlike Embodiment 1, considering that the initial velocity of the imaging medium liquid is relatively high when it first enters the immersion member 121, it is easy for the imaging medium liquid to rush onto the objective lens 21, resulting in more imaging medium liquid contaminating the objective lens 21. At the same time, since some imaging medium liquids are corrosive to metals, in order to reduce the corrosion of the metal parts of the objective lens 21 by the imaging medium liquid, the groove 122 inside the immersion member 121 is provided with a connector for the imaging medium liquid in the reservoir 11 to flow in. An isolation plate 124 is provided at the position between the connector and the inlet 123 in the groove 122 to limit the flow rate of the liquid flowing out of the connector. As shown in the figure, the isolation plate 124 can be an arc-shaped structure. When the imaging medium liquid rushes out of the connector, the isolation plate 124 can prevent the imaging medium liquid from directly rushing onto the objective lens 21.

[0056] Meanwhile, to prevent the imaging medium liquid level in the groove 122 inside the immersion member 121 from being too high, causing the imaging medium liquid to come into contact with the metal part on the objective lens 21, and also to ensure a stable flow rate of the imaging medium liquid in the gap between the objective lens 21 and the socket 123, an opening is provided on the outside of the groove 122 to allow the imaging medium liquid to flow into the water tank 131. When the flow rate of the imaging medium liquid entering the groove 122 is greater than the flow rate of the imaging medium liquid flowing into the socket 123, resulting in excessive accumulation of imaging medium liquid inside the groove 122, the excess imaging medium liquid can be drained into the water tank 131 through this opening.

[0057] Furthermore, in order to provide a stable flow rate of imaging medium liquid into the socket 123, a liquid-blocking ring is provided in the groove 122 near the socket 123, and the surface of the liquid-blocking ring is provided with an opening to allow the imaging medium liquid to flow into the socket 123.

[0058] Example 3:

[0059] Based on Example 2, to ensure that the cut debris does not contaminate too much imaging medium liquid and affect the operation of the debris collection structure 33, the cutting module 3 includes a scraping unit 34 that can scrape off residual imaging medium liquid on the sample surface before the sample is cut. As shown in the figure, the scraping unit 34 is located between the objective lens 21 and the cutting structure 31. The scraping unit 34 includes a flexible scraping strip that can contact the sample surface and a fixing clamp that fixes the flexible scraping strip away from the sample surface, which is intended to clean the residual imaging medium liquid on the sample surface.

[0060] To collect sample debris cut by the cutting structure 31, the debris collection structure 33 includes a suction head 331 located at the cutting edge of the cutting structure 31, a debris filter device 332 connected to the suction head 331 via a pipe, and a negative pressure fan 333 that provides suction to the suction head 331. The suction head 331 and the pipe connecting the debris filter device 332 are both made of transparent material that can withstand negative pressure. The suction head 331 can be structured as shown in the attached figure, which aims to achieve sufficient collection of debris generated during cutting.

[0061] Based on the above embodiments, a cross-sectional immersion imaging method suitable for large-sample, high-resolution three-dimensional imaging is now disclosed, comprising the following steps:

[0062] S1. Fix the sample containing the biological tissue in the water tank 131 and pour the imaging medium into the reservoir 11.

[0063] S2. The three-dimensional translation stage 32 drives the water tank 131 to move, so that the sample is moved directly below the objective lens 21. The control valve and water pump in the imaging medium liquid immersion circulation module 1 are opened, so that the imaging medium liquid in the reservoir 11 circulates between the immersion member 121 and the reservoir 11. At this time, the imaging medium liquid forms a stable imaging medium immersion environment between the objective lens and the sample.

[0064] Meanwhile, before the imaging medium liquid in the water tank 131 enters the reservoir 11, the filter structure 132 filters the sample debris in the imaging medium liquid.

[0065] S3. The water tank 131 and the sample are moved by the three-dimensional translation stage 32, and the illumination imaging module 2 realizes the imaging and recording of the entire sample surface in the imaging medium immersion environment.

[0066] S4. After recording is completed, the control valve and water pump of the pipeline in the imaging medium immersion circulation module 1 are closed, and the three-dimensional translation stage 32 drives the water tank 131 to move as a whole. At this time, the sample leaves the bottom of the objective lens 21, the negative pressure fan 333 in the debris collection structure 33 is started, and the scraping unit 34 scrapes the residual imaging medium liquid on the sample into the water tank 131. Then the sample comes into contact with the cutting edge of the cutting structure 31, and the cutting structure 31 cuts off the sample surface that has been recorded by the illumination imaging module 2, exposing a new sample cross-section. At this time, the debris collection structure 33 collects the cut sample debris.

[0067] S5. Turn off the negative pressure fan 333, and the three-dimensional translation stage 32 will move the sample back to the bottom of the objective lens 21. Repeat steps S2 to S4.

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0069] In the description of this technology, it should also be noted that, unless otherwise explicitly specified and limited, the terms "set," "install," "connect," and "link" should be interpreted broadly. For example, they can refer to fixed connections, detachable connections, or integral connections; they can refer to mechanical connections or electrical connections; they can refer to direct connections or indirect connections through an intermediate medium; and they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this technology based on the specific circumstances. 。

Claims

1. A cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging, characterized in that, include: The imaging medium liquid wetting circulation module (1), the illumination imaging module (2), and the cutting module (3) are included. The imaging medium liquid immersion circulation module (1) includes a reservoir (11) configured to store imaging medium liquid, an immersion assembly (12) connected to the reservoir (11) via a pipe, a medium liquid recovery structure (13) connected to the immersion assembly (12) and the reservoir (11) via a pipe, and a water pump. The imaging medium can complete a full cycle between the reservoir (11), the medium recovery structure (13), and the wetting assembly (12), and the pump can power this cycle. The illumination imaging module (2) is located directly above the sample immersion component (12) and can take pictures of the sample after it has been immersed in the imaging medium liquid. When the imaging medium liquid flows through the immersion component (12) to the sample surface, it can form a stable imaging medium liquid immersion environment between the illumination imaging module (2) and the sample.

2. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 1, characterized in that: The illumination imaging module (2) includes an objective lens (21) positioned above the sample immersion assembly (12) to focus illumination light and collect sample light signals, and an illumination and imaging optical path (22) capable of providing stable illumination and imaging the sample light signals collected by the objective lens (21).

3. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 2, characterized in that: The cutting module (3) includes a cutting structure (31) capable of cutting the sample surface, a three-dimensional translation stage (32) capable of moving the sample as a whole to the cutting structure (31), and a debris collection structure (33) capable of collecting the sample fragments cut off by the cutting structure (31).

4. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 3, characterized in that: The immersion assembly (12) includes an immersion member (121), a groove (122) provided in the immersion member (121) for the inflow of imaging medium liquid, and a socket (123) provided in the groove (122) for the objective lens (21) to be adapted. After the objective lens (21) enters the socket (123), a gap is left between it and the inside of the socket (123) for the imaging medium liquid to flow down.

5. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 4, characterized in that: The medium liquid recovery structure (13) includes a water tank (131) capable of accommodating the sample and a filter structure (132) located between the water tank (131) and the reservoir (11). The filter structure (132) is connected to the water tank (131) and the reservoir (11) via a pipe. The pipe structure in the imaging medium liquid immersion circulation module (1) is equipped with a control valve that can control its opening and closing. When the imaging medium liquid in the water tank (131) enters the reservoir (11), the filter structure (132) can filter the sample debris present in the imaging medium liquid. The water tank (131) is located on the surface of the movable end of the three-dimensional translation stage (32). The water tank (131) can be driven by the three-dimensional translation stage (32) to perform three-dimensional motion. The water tank (131) is equipped with a limiting structure that can limit the position of the sample in the water tank (131).

6. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 5, characterized in that: The cutting module (3) includes a scraping unit (34) that can scrape off residual imaging medium liquid on the sample surface before the sample is cut. The scraping unit (34) is located between the objective lens (21) and the cutting structure (31). The debris collection structure (33) includes a suction head (331) located at the cutting edge of the cutting structure (31), a debris filter device (332) connected to the suction head (331) through a pipe, and a negative pressure fan (333) that provides suction to the suction head (331). The suction head (331) and the pipe connected to the debris filter device (332) are both made of transparent material that can withstand negative pressure.

7. The cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to claim 6, characterized in that: The immersion member (121) has a groove (122) inside which a connector is provided for the imaging medium liquid in the reservoir (11) to flow in. A partition plate (124) is provided between the connector and the socket (123) in the groove (122) to limit the flow rate of the liquid flowing out of the connector. An opening is provided on the outside of the groove (122) for the imaging medium liquid to flow into the water tank (131). A liquid-blocking ring is provided in the groove (122) near the socket (123). The surface of the liquid-blocking ring has an opening for the imaging medium liquid to flow into the socket (123).

8. A cross-section immersion imaging method suitable for large sample high resolution three-dimensional imaging, which uses the cross-section immersion imaging system suitable for large sample high resolution three-dimensional imaging according to any one of claims 1 to 7, characterized in that, Includes the following steps: S1. Fix the sample containing the embedded biological tissue in the water tank (131) and pour the imaging medium into the reservoir (11); S2. The water tank (131) is moved by the three-dimensional translation stage (32) to move the sample to the bottom of the objective lens (21). The control valve and water pump in the imaging medium liquid immersion circulation module (1) are opened to allow the imaging medium liquid in the reservoir (11) to circulate between the immersion part (121) and the reservoir (11). At this time, the imaging medium liquid forms a stable imaging medium immersion environment between the objective lens and the sample. S3. The water tank (131) and the sample are moved by the three-dimensional translation stage (32), and the entire sample surface is photographed and recorded by the illumination imaging module (2) in the imaging medium immersion environment. S4. After recording is completed, the three-dimensional translation stage (32) moves the water tank (131) as a whole. At this time, the sample leaves the bottom of the objective lens (21) and comes into contact with the cutting edge of the cutting structure (31). The cutting structure (31) cuts off the sample surface that has been recorded by the illumination imaging module (2) to expose a new sample cross-section. S5. The three-dimensional translation stage (32) moves the sample back to the bottom of the objective lens (21), and steps S2 to S4 are repeated.

9. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 8, characterized in that: In step S2, before the imaging medium liquid in the water tank (131) enters the reservoir (11), the filter structure (132) filters the sample debris in the imaging medium liquid.

10. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 9, characterized in that: In step S4, after the illumination imaging module (2) records the imaging results of the sample surface, before the three-dimensional translation stage (32) moves the water tank (131) as a whole, the control valves and water pumps in the pipeline of the imaging medium liquid immersion circulation module (1) are closed.

11. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 10, characterized in that: In step S4, after the illumination imaging module (2) records the imaging results of the sample surface, the three-dimensional translation stage (32) drives the water tank (131) and the sample as a whole to move before being cut by the cutting structure (31), the negative pressure fan (333) in the debris collection structure (33) is started.

12. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 11, characterized in that: In step S4, after the cutting structure (31) has finished cutting the sample, the negative pressure fan (333) is turned off.

13. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 12, characterized in that: In step S4, before the portion of the image that has been recorded is cut off by the cutting structure (31), the scraping unit (34) can scrape the residual imaging medium liquid on the sample into the water tank (131).

14. The section-infiltration imaging method suitable for large sample high resolution three-dimensional imaging according to claim 13, characterized in that: The three-dimensional translation stage (32) can raise the overall height of the water tank (131) after each cutting of the sample surface by the cutting structure (31) for the next cutting of the sample surface.

Images