Section infiltration type imaging system and method suitable for large-sample high-resolution three-dimensional imaging

Abstract

The invention discloses a section infiltration type imaging system suitable for large-sample high-resolution three-dimensional imaging. The section infiltration type imaging system comprises an imaging medium liquid infiltration circulation module, an illumination imaging module and a cutting module. Through the imaging medium liquid infiltration circulating module, a stable imaging medium liquid infiltration environment is only formed between the shooting unit and a sample in the process of using a mechanical cutting imaging method when three-dimensional imaging of a biological large sample is carried out; compared with an existing method of soaking a sample embedded with biological tissues in imaging medium liquid, the method provided by the invention has the advantages that high-resolution imaging can be realized only by using a small amount of imaging medium liquid capable of forming an imaging medium liquid infiltration environment; the condition that the cutting performance is affected due to sample deformation caused by long-term soaking of the sample in the high-refractive-index imaging medium liquid is effectively avoided, the amount of the imaging medium liquid used in the water tank is further reduced, the use cost is reduced, and meanwhile the load requirement for the three-dimensional translation table is lowered.

Description

Technical Field

[0001] The present application relates to the fields of research and medicine, and specifically to a cross-sectional immersion imaging system and method suitable for high-resolution three-dimensional imaging of large samples. Background Art

[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 mouse whole brain and monkey brain. These systems can be roughly divided into two types of imaging methods: tissue transparency-based and mechanical cutting-based.

[0003] Among them, the imaging method based on tissue transparency is to make biological tissue transparent to increase the depth of optical imaging and use light sheet fluorescence microscopy for rapid imaging. This method is suitable for large-scale rapid imaging, but due to the problem of residual refractive index mismatch in the deep layer of the sample, the imaging resolution and quality will decrease significantly as the imaging depth increases.

[0004] The imaging method based on mechanical cutting combines optical tomography technology with precision cutting technology, embedding biological tissue in a specific medium to form a sample to achieve a hardness that meets the requirements of precision cutting. When imaging the sample, the sample is placed on a three-dimensional translation stage, and the objective lens focuses the illumination light on the shallow part of the sample section and collects the imaging light. After the sample is driven by the three-dimensional translation stage to obtain the tomographic image of the shallow part of the entire section through the illumination and imaging module, the imaged part of the sample surface is cut off with a tool, and the three-dimensional imaging of large centimeter-sized samples can be achieved by continuously repeating the "section imaging-surface cutting" process. The advantage of this method is that the imaging focal plane is always in the shallow layer of the sample with very low light scattering and light absorption, which can ensure the overall uniform imaging quality of large-size samples and obtain high-resolution three-dimensional imaging data.

[0005] In the imaging method based on mechanical cutting, in order to improve the imaging resolution, it is often necessary to use an objective lens adapted to high refractive index imaging medium liquid, such as a water objective lens, an oil objective lens, and an objective lens adapted to other high refractive index imaging medium liquid. When using this type of objective lens, the sample is generally fixed in a water tank filled with imaging medium liquid. However, this solution will bring a series of problems, including:

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

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

[0008] (3) When the water tank is filled with imaging medium liquid, the weight increases significantly, resulting in an increase in the load of the 3D translation stage, affecting the dynamic positioning accuracy of the 3D translation stage, or requiring the use of a more expensive high-load 3D translation stage. Summary of the invention

[0009] To solve the above problems, this application provides the following technical solutions:

[0010] A cross-sectional immersion imaging system suitable for high-resolution three-dimensional imaging of large samples, comprising: 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 liquid reservoir configured to store imaging medium liquid, an immersion assembly connected to the liquid reservoir through a pipeline, a medium liquid recovery structure connected to the immersion assembly and the liquid reservoir through a pipeline, and a water pump, wherein the imaging medium liquid can perform a complete cycle between the liquid reservoir, the medium liquid recovery structure, and the immersion assembly, and the water pump can provide power for the cycle;

[0012] The illumination imaging module is arranged directly above the sample infiltration component and can photograph the sample after being infiltrated with the imaging medium liquid. When the imaging medium liquid flows through the infiltration component onto the sample surface, a stable imaging medium liquid infiltration environment can be formed between the illumination imaging module and the sample.

[0013] Based on the above technical solution, the present application can also be improved as follows.

[0014] Furthermore, the illumination imaging module includes an objective lens disposed above the sample immersion component for focusing the illumination light and collecting the sample light signal, and 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 driving the sample as a whole to move to the cutting structure, and a debris collection structure capable of collecting sample fragments cut off by the cutting structure.

[0016] Furthermore, the immersion component includes an immersion piece, a groove in the immersion piece for imaging medium liquid to flow in, and a socket in the groove adapted to the objective lens. After the objective lens enters the socket, a gap is left between the objective lens and the inside of the socket for the imaging medium liquid to flow down.

[0017] Furthermore, the medium liquid recovery structure includes a water tank capable of accommodating samples, and a filtering structure arranged between the water tank and the liquid reservoir. The filtering structure is connected to the water tank and the liquid reservoir through a pipeline. The pipeline structure in the imaging medium liquid infiltration circulation module is provided with a control valve capable of controlling its on and off. When the imaging medium liquid in the water tank enters the liquid reservoir, the filtering 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. The water tank can be driven by the three-dimensional translation stage to perform three-dimensional movement. A limiting structure capable of limiting the position of the sample in the water tank is provided in the water tank.

[0018] Furthermore, the cutting module includes a scraping unit capable of scraping off residual imaging medium liquid on the surface of the sample before the sample is cut, and the scraping unit is located between the objective lens and the cutting structure. The debris collection structure includes a suction head arranged at the cutting edge of the cutting structure, a debris filtering device connected to the suction head through a pipeline, and a negative pressure fan providing suction to the suction head. The suction head and the pipeline connected to the debris filtering device are both made of transparent materials that can withstand negative pressure.

[0019] Furthermore, a docking port is provided in the inner groove of the immersion component for the imaging medium liquid in the liquid reservoir to flow into, an isolation plate capable of limiting the flow rate of liquid flowing out of the docking port is provided between the docking port and the socket in the groove, an opening is provided on the outer side of the groove for the imaging medium liquid to flow into the water tank, a liquid blocking ring is provided at a position near the socket in the groove, and an opening is provided on the surface of the liquid blocking ring for the imaging medium liquid to flow into the inside of the socket.

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

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

[0022] S2, the water tank is driven to move by the three-dimensional translation stage, so that the sample is moved to the right below the objective lens, and the control valve and water pump in the imaging medium liquid immersion circulation module are opened to allow the imaging medium liquid in the reservoir to circulate between the immersion member 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 driven to move by the three-dimensional translation stage, and the illumination imaging module is used to shoot and record the entire sample surface in the imaging medium immersion environment;

[0024] S4, after recording is completed, the three-dimensional translation stage drives the water tank to move as a whole, at which time the sample leaves the bottom of the objective lens and contacts the blade of the cutting structure, and the cutting structure cuts off the sample surface whose image has been recorded by the illumination imaging module, exposing a new sample section;

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

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

[0027] Furthermore, in step S4, after the illumination imaging module records the imaging result of the sample surface and before the three-dimensional translation stage drives the water tank to move as a whole, the control valve and the water pump of the pipeline in the imaging medium liquid infiltration circulation module are closed.

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

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

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

[0031] Furthermore, the three-dimensional translation stage can drive the entire height of the water tank to rise after the cutting structure cuts the sample surface each time, so as to facilitate the next cutting of the sample surface.

[0032] The details of one or more embodiments of the present application are set forth in the following drawings and description. Other features, objects, and advantages of the present application will become apparent from the description, drawings, and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to better describe and illustrate the embodiments and or examples of the inventions disclosed herein, reference may be made to one or more drawings. The additional details 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, and the best mode of these inventions currently understood.

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

[0035] Figure 2 is a perspective schematic diagram of an infiltration assembly according to one or more embodiments;

[0036] Figure 3 is a schematic diagram of a front cross-sectional structure of an infiltration component according to one or more embodiments;

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

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

[0039] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0040] 1. Imaging medium liquid infiltration circulation module; 11. Liquid reservoir; 12. Infiltration component; 121. Infiltration part; 122. Groove; 123. Socket; 124. Isolation plate; 13. Medium liquid recovery structure; 131. Water tank; 132. Filtering structure; 2. Illumination 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 filtering device; 333. Negative pressure fan; 34. Wiping unit. DETAILED DESCRIPTION

[0041] See also Figure 1 The present 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.

[0042] Among them, the imaging medium liquid infiltration circulation module 1 includes a reservoir 11 configured to store imaging medium liquid, an infiltration component 12 connected to the reservoir 11 through a pipeline, a medium liquid recovery structure 13 connected to the infiltration component 12 and the reservoir 11 through a pipeline, and a water pump. The imaging medium liquid can perform a complete cycle between the reservoir 11, the medium liquid recovery structure 13, and the infiltration component 12. The water pump can provide power for this cycle. A control valve that can control the on-off of the pipeline is provided on the pipeline in the imaging medium liquid infiltration circulation module 1. In order to ensure smooth circulation between the infiltration component 12, the reservoir 11 and the water pump, the three can be interconnected through a pipeline. Through the uninterrupted circulation of the imaging medium liquid, a stable flow of imaging medium liquid can be provided for the shooting of the sample when the sample is not immersed in the imaging medium liquid.

[0043] The imaging medium liquid can be selected from water, aqueous solution containing dyes, glycerin, immersion oil, and other composite components of high refractive index imaging medium liquids according to imaging requirements.

[0044] The lighting imaging module 2 is an optical and electronic device commonly used in mechanical cutting imaging methods in the prior art, including an objective lens 21 arranged above the sample immersion component 12 for focusing the illumination light and collecting the sample light signal, and an illumination and imaging optical path 22 capable of providing stable illumination and imaging the sample light signal collected by the objective lens. When the imaging medium liquid passes through the immersion component 12 and flows onto the sample surface, a stable imaging medium liquid immersion environment can be formed between the objective lens 21 and the sample.

[0045] The medium immersion environment here refers to the liquid environment formed between the position where the sample needs to be photographed and the objective lens 21 when the imaging medium liquid has a stable flow rate due to its own tension. The imaging medium liquid can form a stable imaging medium liquid immersion environment by controlling the distance between the socket 123 and the objective lens 21, the distance between the lower surface of the immersion component 12 and the sample surface, and the microstructure of the lower surface of the immersion component 12.

[0046] The cutting module 3 includes a cutting structure 31 capable of cutting the surface of the sample, a three-dimensional translation stage 32 capable of driving the sample as a whole to move to the cutting structure 31, and a debris collection structure 33 capable of collecting sample fragments cut off by the cutting structure 31. The three-dimensional translation stage 32 is a common electrical device in the prior art, which can drive the water tank 131 on the three-dimensional translation stage 32 to move in the horizontal, longitudinal and vertical directions through the cooperation of the motor and the mechanical structure. During the sample cutting process, the three-dimensional translation stage 32 can adjust the height of the water tank 131 in accordance with the lowered height of the sample after being cut, to ensure that the cutting structure 31 can cut the unphotographed part of the sample.

[0047] The three-dimensional translation stage 32 can realize imaging of a large sample surface. Since the imaging field of the objective lens 21 is small, for a large sample surface, the three-dimensional translation stage 32 needs to drive the sample to expand the imaging range through scanning imaging.

[0048] Compared with the prior art, in the above technical solution, only the upper section of the sample is in contact with the imaging medium liquid, thus avoiding deformation of the sample and affecting cutting performance due to long-term immersion of the sample as a whole in the imaging medium liquid. 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.

[0049] At the same time, due to the reduction in the amount of imaging medium liquid used, the requirements for the load capacity of the three-dimensional translation stage 32 are greatly reduced, thereby reducing the system hardware cost.

[0050] In order to explain the above technical scheme in detail, the following embodiments are listed. What is described below is only a specific implementation mode of the present application, but the protection scope of the present application is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present application within the technical scope disclosed in the present application, which should be covered by the protection scope of the present application.

[0051] Embodiment 1:

[0052] On the basis of the above technical solution, the immersion component 12 includes an immersion member 121, a groove 122 provided in the immersion member 121 for the imaging medium liquid to flow in, and a socket 123 provided in the groove 122 and adapted to the objective lens 21. After the objective lens 21 enters the interior of the socket 123, a gap is left between it and the interior of the socket 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 an annular structure as shown in the figure. The immersion member 121 as a whole can be sleeved on the surface of the objective lens 21. When photographing the sample, 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 infiltration environment between the objective lens 21 and the sample.

[0053] On the basis of the above, in order to collect and reuse the imaging medium liquid flowing out of the immersion element 121, the medium liquid recovery structure 13 includes a water tank 131 capable of accommodating samples, and a filtering structure 132 arranged between the water tank 131 and the liquid reservoir 11. The filtering structure 132 is connected to the water tank 131 and the liquid reservoir 11 through a pipeline structure. When the imaging medium liquid in the water tank 131 enters the liquid reservoir 11, the filtering structure 132 can filter the sample debris present in the imaging medium liquid. The filtering structure 132 can be a common structure composed of a device shell and an internal filter material, which is intended to filter out the sample debris contained in the imaging medium liquid entering the liquid reservoir 11 from the water tank 131, so as to ensure the cleanliness of the imaging medium liquid stored in the liquid reservoir 11 and avoid affecting the imaging effect of the subsequent lighting imaging module 2.

[0054] Preferably, the bottom of the water tank 131 is an inclined surface, and the bottom of the inclined surface is connected to the pipe leading to the filtering structure 132. Through the design of the inclined surface at the bottom of the water tank 131, 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 mouth and be collected by the water pump into the liquid reservoir 11, so as to reduce the imaging medium liquid accumulated in the water tank 131.

[0055] Embodiment 2:

[0056] Different from Example 1, considering that the initial velocity of the imaging medium liquid is relatively fast when it just enters the immersion member 121, it is easy for the imaging medium liquid to rush onto the objective lens 21, causing more imaging medium liquid to be contaminated onto the objective lens 21. At the same time, since some imaging medium liquids are corrosive to metals to a certain extent, in order to reduce the corrosion of the imaging medium liquid on the metal part of the surface of the objective lens 21, a docking port for the imaging medium liquid in the liquid reservoir 11 to flow into is provided in the groove 122 inside the immersion member 121, and an isolation plate 124 capable of limiting the flow rate of the liquid flowing out of the docking port is provided at a position between the docking port and the socket 123 in the groove 122. As shown in the figure, the isolation plate 124 can be an arc-shaped structure. When the imaging medium liquid rushes out of the docking port, the isolation plate 124 can prevent the imaging medium liquid from directly rushing onto the objective lens 21.

[0057] At the same time, in order to prevent the liquid level of the imaging medium liquid in the groove 122 inside the immersion member 121 from being too high, causing the imaging medium liquid to contact the metal part on the objective lens 21, and also to ensure the stability of the 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 outer side of the groove 122 for 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 the imaging medium liquid inside the groove 122, the excess imaging medium liquid can be discharged into the water tank 131 through the opening here.

[0058] Furthermore, in order to provide imaging medium liquid with a stable flow rate into the socket 123 , a liquid blocking ring is provided in the groove 122 near the socket 123 , and an opening is provided on the surface of the liquid blocking ring for the imaging medium liquid to flow into the socket 123 .

[0059] Embodiment 3:

[0060] On the basis of Example 2, in order to ensure that the cut debris will not be contaminated with too much imaging medium liquid to affect the operation of the debris collection structure 33, the cutting module 3 includes a scraping unit 34 that can scrape off the 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 for fixing the surface of the flexible scraping strip away from the sample at one end, so as to clean the residual imaging medium liquid on the sample surface.

[0061] In order to collect the sample debris cut off by the cutting structure 31, the debris collection structure 33 includes a suction head 331 arranged at the cutting edge of the cutting structure 31, a debris filtering device 332 connected to the suction head 331 through a pipeline, and a negative pressure fan 333 providing suction to the suction head 331. The suction head 331 and the pipeline connected to the debris filtering device 332 are both made of transparent materials that can withstand negative pressure. The suction head 331 can be structured as shown in the accompanying drawing, aiming to fully collect the debris generated by cutting.

[0062] In combination with the above embodiments, a cross-sectional immersion imaging method suitable for high-resolution three-dimensional imaging of large samples is now disclosed, comprising the following steps:

[0063] S1 . Fix the sample embedded with biological tissue in the water tank 131 , and pour the imaging medium liquid into the liquid reservoir 11 .

[0064] S2. The water tank 131 is driven to move 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 liquid reservoir 11 to circulate between the immersion member 121 and the liquid reservoir 11. At this time, the imaging medium liquid forms a stable imaging medium immersion environment between the objective lens and the sample.

[0065] At the same time, before the imaging medium liquid in the water tank 131 enters the liquid reservoir 11 , the filtering structure 132 filters the sample debris in the imaging medium liquid.

[0066] S3. The water tank 131 and the sample are moved by the three-dimensional translation stage 32, and the illumination imaging module 2 shoots and records the entire sample surface in the imaging medium immersion environment.

[0067] S4. After recording is completed, the control valve and water pump of the pipeline in the imaging medium liquid infiltration 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, and the negative pressure fan 333 in the debris collection structure 33 is started. The scraping unit 34 scrapes the imaging medium liquid remaining on the sample into the water tank 131. Then the sample contacts the blade of the cutting structure 31, and the cutting structure 31 cuts off the sample surface whose image has been recorded by the illumination imaging module 2 to expose a new sample section. At this time, the debris collection structure 33 collects the cut sample debris.

[0068] S5. Turn off the negative pressure fan 333, and the three-dimensional translation stage 32 drives the sample back to the bottom of the objective lens 21, and repeat steps S2 to S4.

[0069] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0070] In the description of this technology, it is also necessary to explain that, unless otherwise clearly specified and limited, the terms "set", "install", "connect", and "connect" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in this technology can be understood according to specific circumstances.

Claims

1. A cross-sectional immersion imaging system suitable for high-resolution three-dimensional imaging of large samples, characterized in that: include: An imaging medium liquid infiltration circulation module (1), an illumination imaging module (2) and a cutting module (3); The imaging medium liquid immersion circulation module (1) comprises a liquid reservoir (11) configured to store imaging medium liquid, an immersion component (12) connected to the liquid reservoir (11) via a pipeline, a medium liquid recovery structure (13) connected to the immersion component (12) and the liquid reservoir (11) via a pipeline, and a water pump; The imaging medium liquid can perform a complete cycle between the liquid reservoir (11), the medium liquid recovery structure (13), and the immersion component (12), and the water pump can provide power for this cycle; The illumination imaging module (2) is arranged directly above the sample infiltration component (12) and is capable of photographing the sample after being infiltrated with the imaging medium liquid. When the imaging medium liquid passes through the infiltration component (12) and flows onto the sample surface, a stable imaging medium liquid infiltration environment can be formed between the illumination imaging module (2) and the sample.

2. A cross-sectional immersion imaging system suitable for large sample high-resolution three-dimensional imaging according to claim 1, characterized in that: The illumination imaging module (2) comprises an objective lens (21) arranged above the sample immersion component (12) for focusing illumination light and collecting 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-sectional immersion imaging system suitable for high-resolution three-dimensional imaging of large samples according to claim 2, characterized in that: The cutting module (3) comprises a cutting structure (31) capable of cutting the surface of a sample, a three-dimensional translation platform (32) capable of driving the entire sample to move to the cutting structure (31), and a debris collection structure (33) capable of collecting sample fragments cut off by the cutting structure (31).

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

5. The cross-sectional 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) comprises a water tank (131) capable of accommodating a sample, and a filtering structure (132) arranged between the water tank (131) and the liquid reservoir (11); the filtering structure (132) is connected to the water tank (131) and the liquid reservoir (11) through a pipeline; a control valve capable of controlling the on-off of the pipeline structure in the imaging medium liquid infiltration circulation module (1) is provided; when the imaging medium liquid in the water tank (131) enters the liquid reservoir (11), the filtering structure (132) can filter 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 movement; and a limiting structure capable of limiting the position of the sample in the water tank (131) is provided in the water tank (131).

6. The cross-sectional immersion imaging system suitable for high-resolution three-dimensional imaging of large samples according to claim 5, characterized in that: The cutting module (3) comprises a scraping unit (34) capable of scraping off residual imaging medium liquid on the surface of the sample 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) comprises a suction head (331) arranged at the cutting edge of the cutting structure (31), a debris filtering device (332) connected to the suction head (331) through a pipeline, and a negative pressure fan (333) providing suction force for the suction head (331). The suction head (331) and the pipeline connected to the debris filtering device (332) are both made of a transparent material capable of withstanding negative pressure.

7. The cross-sectional immersion imaging system suitable for large sample high-resolution three-dimensional imaging according to claim 6, characterized in that: A docking port for the imaging medium liquid in the liquid reservoir (11) to flow into is provided in the groove (122) inside the immersion member (121); an isolation plate (124) capable of limiting the flow rate of liquid flowing out of the docking port is provided at a position between the docking port and the insertion port (123) inside the groove (122); an opening for the imaging medium liquid to flow into the water tank (131) is provided outside the groove (122); a liquid blocking ring is provided at a position near the insertion port (123) inside the groove (122); and an opening is provided on the surface of the liquid blocking ring for the imaging medium liquid to flow into the inside of the insertion port (123).

8. A cross-sectional immersion imaging method suitable for large sample high-resolution three-dimensional imaging, using the cross-sectional immersion imaging system suitable for large sample high-resolution three-dimensional imaging according to any one of claims 1 to 7, characterized in that: The steps include: S1, fix the sample embedded with biological tissue in a water tank (131), and pour the imaging medium liquid into the liquid reservoir (11); S2, driving the water tank (131) to move by means of the three-dimensional translation stage (32), so that the sample is moved to the position directly below the objective lens (21), and opening the control valve and the water pump in the imaging medium liquid infiltration circulation module (1), so that the imaging medium liquid in the liquid reservoir (11) circulates between the infiltration member (121) and the liquid reservoir (11), and at this time, the imaging medium liquid forms a stable imaging medium infiltration 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 illumination imaging module (2) is used to capture and record the entire sample surface in the imaging medium immersion environment; S4, after the recording is completed, the three-dimensional translation stage (32) drives the water tank (131) to move as a whole, and at this time the sample leaves the bottom of the objective lens (21) and contacts the blade of the cutting structure (31), and the cutting structure (31) cuts off the sample surface whose image has been recorded by the illumination imaging module (2), thereby exposing a new sample cross section; S5. The three-dimensional translation stage (32) drives the sample back to the bottom of the objective lens (21), and steps S2 to S4 are repeated.

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

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

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

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

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

14. The cross-sectional immersion imaging method suitable for high-resolution three-dimensional imaging of large samples according to claim 13, characterized in that: The three-dimensional translation stage (32) can drive the entire height of the water tank (131) to rise after the cutting structure (31) cuts the sample surface each time, so as to facilitate the next cutting of the sample surface.

Images