Cell growth in-situ visualization device based on X-ray and use method

Abstract

The invention relates to the technical field of cell observation, in particular to an X-ray-based cell growth in-situ visualization device and a use method. A cell growth in-situ visualization device is used for being installed on a mobile platform with a guide rail. The device comprises a sample container and a rotary base, cells needing to be observed are stored in the sample container which is of a cylinder structure and is hollow and free of an upper bottom face, and the inverted-cone-shaped space is formed in the bottom of the interior of the sample container, the cells can be gathered to the tip end of the bottom of the inverted-cone-shaped space, it can be guaranteed that the cells are located in the imaging range all the time, and in-situ observation of the growth process is achieved; x-rays penetrate through the cells to form a phase contrast image, so that the integral definition of the generated image is improved through the imaging mode. Meanwhile, two-dimensional images of the cells at different angles can be obtained on the basis of the rotating base and in combination with an X-ray phase contrast CT imaging method, then three-dimensional images of the cells are constructed, and the microstructure of the cells can be observed more comprehensively in situ.

Description

Technical Field

[0001] The present invention relates to the technical field of cell observation, and in particular to an X-ray-based in-situ visualization device for cell growth and a use method thereof. Background Art

[0002] The statements herein merely provide background information related to the present invention and do not necessarily constitute prior art.

[0003] In situ observation of cell growth is very important for research in biology and medicine. It can provide a direct and detailed understanding of the dynamic changes of cells. Its importance can be reflected in the following aspects:

[0004] (1) Revealing the process of cell development and differentiation: It can monitor cell division, migration and differentiation in real time from embryonic development to maturity. (2) Studying disease mechanisms: Observing the actual growth process of cells in the body helps to reveal the mechanism of disease. (3) Evaluating the effects of drugs and treatments: It can be used to evaluate the effects of drugs or treatments on cell growth and function. (4) Promoting the progress of cell engineering and synthetic biology: It can provide real-time data for cell engineering and synthetic biology, helping scientists design and optimize new cell models and systems.

[0005] In the prior art, in situ observation of the cell growth process is usually carried out using an optical microscope or a fluorescence microscope. When an optical microscope is used, it is limited by the resolution of the microscope and the diffraction limit of light and cannot clearly distinguish the subtle cell structure. When a fluorescence microscope is used, the cells need to be fluorescently labeled with probes and fluorescent agents, which will affect the normal growth of the cells and can only image the labeled cells. At the same time, the fluorescent markers will gradually lose fluorescence, thereby affecting the clarity and accuracy of the image.

[0006] Moreover, when observing with a microscope, the cell sample is placed in a culture dish for observation, and only a two-dimensional image of the cells can be observed; at the same time, when performing high-resolution imaging, the imaging field of view becomes smaller, and the sample to be observed is extremely easy to run out of the field of view, thus affecting the observation of the cells. Summary of the invention

[0007] The technical problem to be solved by the present invention is to provide an X-ray-based in-situ visualization device for cell growth and a method for using the device. Cells are stored in a sample container which is hollow inside, has no upper bottom surface, and has an inverted cone-shaped space at the bottom. High-resolution three-dimensional images of the cell growth process are generated by combining X-ray phase contrast imaging technology and CT methods, thereby observing the cell growth process in situ and clearly observing the cell microstructure without affecting the cell growth.

[0008] The object of the present invention is to provide an X-ray-based in-situ visualization device for cell growth, which is used to be installed on a mobile platform with a guide rail. The device includes a sample container and a rotating base, and the rotating base is nested on the guide rail of the mobile platform; the sample container is a column structure with a hollow interior and no upper bottom surface, and the bottom of the interior is an inverted cone space. An insertion table is provided at the bottom of the outer side of the sample container, and the sample container is plug-connected to the slot of the rotating base through the insertion table.

[0009] As a further technical solution, the sample container is in a vertical state.

[0010] As a further technical solution, the rotating base includes a supporting platform, a lifting rod, a rotating disk and a base, the insert platform includes a circular positioning boss in the middle and square limiting bosses located at both ends of the positioning boss, the slot includes a circular positioning socket in the middle and square limiting sockets located at both ends of the positioning socket, the positioning boss is inserted into the positioning socket, the limiting boss is inserted into the limiting socket, and the insert platform and the slot are made of magnetic materials to attract the sample container and the rotating base; one end of the lifting rod is fixedly connected to the bottom of the supporting platform, and the other end of the lifting rod is fixedly connected to the rotating disk, and the rotating disk is arranged above the base.

[0011] As a further technical solution, the lifting rod is specifically an electric lifting rod.

[0012] As a further technical solution, the base includes a shell, and the four sides of the shell have the same structure; through holes are symmetrically opened on both sides of the bottom end of a single side of the shell, and the two opposite sides of the shell are nested on the guide rail of the mobile platform through the through holes.

[0013] As a further technical solution, the base further includes a motor, which is disposed inside the shell, and a central rotating shaft of the motor is fixedly connected to the bottom of the rotating disk through the shell.

[0014] As a further technical solution, the inverted cone space is specifically an inverted circular cone space.

[0015] As a further technical solution, the sample container is made of polyethylene.

[0016] A method for using an X-ray-based in-situ visualization device for cell growth, the specific steps comprising:

[0017] S1, placing the mobile platform and the detector on the precision translation stage, and the rotating base is nested on the guide rail of the mobile platform using the through hole on the shell;

[0018] S2. delivering the nutrient solution containing the cells into the sample container through a syringe, and ensuring that the nutrient solution is stored in the inverted cone space of the sample container;

[0019] S3, using the insertion platform on the side of the sample container to plug and connect the sample container with the slot of the rotating base, and ensure that the sample container is in a vertical state;

[0020] S4, adjusting the position of the sample container by means of the lifting rod and the moving platform so that the position of the nutrient solution in the sample container and the detector are on the same horizontal line;

[0021] S5. Finally, the nutrient solution and cells in the mobile platform are horizontally irradiated with X-rays, and the rotating disk is used to drive the sample container to rotate at a constant speed; the detector receives the X-rays that have irradiated the nutrient solution and cells, and converts them into electrical signals and transmits them to the computer. The computer uses phase contrast CT to perform three-dimensional imaging and performs observation and analysis based on the three-dimensional images.

[0022] Beneficial effects of one or more of the above technical solutions:

[0023] (1) This embodiment uses a sample container that is hollow inside, has no upper bottom surface, and has an inverted cone-shaped space at the bottom to store the cells to be observed. The cells can be gathered at the tip of the bottom of the inverted cone-shaped space, thereby reducing the generation of artifacts and improving the overall clarity of the generated image by utilizing the phase change after the X-rays pass through the cells. At the same time, it can ensure that the cells move normally during their normal growth and development process, thereby avoiding the problem of cells leaving the imaging field of view.

[0024] (2) This embodiment utilizes a rotating base to fix and rotate the sample container at a constant speed. On the premise of fixing and supporting the sample container, the sample container is driven to rotate by the rotating disk, so that images of cells at different angles can be obtained, thereby accurately constructing a three-dimensional image of the cells and achieving a more comprehensive observation of the microscopic structure of the cells.

[0025] (3) This embodiment uses polyethylene as the material of the sample container, taking advantage of its own transparency and low X-ray absorption capacity, to help generate clearer images without significantly affecting the transmission and scattering of X-rays, thereby facilitating clearer in situ observation of the fine structure of cells. At the same time, imaging is performed based on X-ray phase contrast imaging technology and CT, without the need to label cells, thereby avoiding the impact on cell growth and overcoming the disadvantage of fluorescent labeling microscopic imaging that the image clarity and accuracy are affected by the gradual loss of fluorescence.

[0026] (4) By using the inverted cone-shaped space inside the sample container in this embodiment to place the nutrient solution containing cells, the cells can maintain a certain three-dimensional state, thereby generating a three-dimensional image of the cells, thereby enabling more detailed in-situ observation of the cell structure and other information. At the same time, even if the cells move normally in the nutrient solution, they still exist in the inverted cone-shaped space in the sample container and are within the imaging range, which does not affect the generation of the image. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] The drawings in the specification that constitute a part of this application are used to provide further understanding of the present application. In order to facilitate understanding, the proportions between the structures of each part are adjusted. The schematic embodiments of the present application and their descriptions are used to explain the present application and do not constitute a limitation on the present application.

[0028] Figure 1 This is a structural diagram of the connection between the sample container, the rotating base and the moving platform in the X-ray-based in-situ visualization device for cell growth of the present invention.

[0029] Figure 2 It is a structural diagram of the sample container in the X-ray-based in-situ visualization device for cell growth of the present invention.

[0030] Figure 3 It is a structural diagram of the rotating base in the X-ray-based in-situ visualization device for cell growth of the present invention.

[0031] Figure 4 This is a structural diagram of the connection between the sample container and the rotating base in the X-ray-based in-situ visualization device for cell growth of the present invention.

[0032] Figure 5 The schematic diagram is a method for using the X-ray based in situ visualization device for cell growth of the present invention.

[0033] Among them, 1. sample container, 1-1. hollow area, 1-2. insert table, 2. rotating base, 2-1. support table, 2-2. lifting rod, 2-3. rotating disk, 2-4. base, 2-5. shell, 2-6. slot, 2-7. through hole, 3. moving platform, 4. X-ray, 5. precision translation stage, 6. detector, 7. computer. DETAILED DESCRIPTION

[0034] The following is combined with Figure 1-5 , clearly and completely describe the technical solutions in the embodiments of the present invention.

[0035] Embodiment 1

[0036] Reference Figure 1, an X-ray-based in-situ visualization device for cell growth, the device is used to be installed on a moving platform 3 containing a guide rail; wherein the device includes a sample container 1 and a rotating base 2.

[0037] Reference Figure 2 The sample container 1 is a cylindrical structure with a hollow interior and no upper bottom surface, and the bottom of the interior is an inverted cone-shaped space. In this embodiment, the sample container 1 adopts a cylindrical structure, and the upper bottom surface is set to an open state, and a hollow area 1-1 is set inside the sample container 1; specifically, the inverted cone-shaped space can be understood as an inverted cone-shaped space excavated on the bottom surface of the sample container 1, and the space is used to store the nutrient solution containing cells.

[0038] In the prior art, in situ observation of cells usually involves placing a culture medium containing cells in a culture dish and observing them through an optical microscope or a fluorescence microscope. However, in this method, the cells are all in an adherent growth state (close to the bottom or side of the culture dish), resulting in only images of extremely thin cells that are close to a two-dimensional state being generated during the imaging process, which is different from the three-dimensional structure of the cells in their true state; at the same time, the in situ observation of the three-dimensional structure of cells is limited by factors such as changes in biological activity caused by cell attachment and possible unreal interactions and spatial distribution between cells.

[0039] When using an optical microscope for observation, it is impossible to clearly distinguish subtle cell structures due to the limitations of the microscope's resolution and the diffraction limit of light; when the microscope's resolution is increased, the microscope's imaging field of view will decrease, and coupled with the normal movement of cells during their normal growth and development, the cells will move out of the imaging field of view, ultimately making it impossible to quickly and accurately observe the cells in situ.

[0040] By using the inverted cone-shaped space inside the sample container 1 in this embodiment to place the nutrient solution containing cells, the cells can maintain a certain three-dimensional state, so that a three-dimensional image of the cells can be generated, so that the structure of the cells and other information can be observed in more detail in situ. At the same time, even if the cells move normally in the nutrient solution, they still exist in the inverted cone-shaped space in the sample container 1 and are within the imaging range, which will not affect the generation of the image.

[0041] In order to further fit the state of the cells and avoid damage to the cells, the inverted cone space in this embodiment is specifically an inverted cone space. At the same time, an inserting platform 1-2 is provided at the bottom of the outer side of the sample container 1, and the sample container 1 is plugged and connected with the slot 2-6 of the rotating base 2 through the inserting platform 1-2, and the sample container 1 is arranged on the rotating base 2 in a vertical state (such as Figure 4 shown).

[0042] Reference Figure 3, the rotating base 2 includes a support table 2-1, a lifting rod 2-2, a rotating disk 2-3 and a base 2-4. Specifically, the inserting table 1-2 includes a circular positioning boss in the middle and square limiting bosses at both ends of the positioning boss, and the slot 2-6 includes a circular positioning socket in the middle and square limiting sockets at both ends of the positioning socket. The positioning boss is inserted into the positioning socket to achieve coaxial positioning of the sample container 1 and the rotating base 2, and the limiting boss is inserted into the limiting socket to achieve axial limiting of the sample container 1 and the rotating base 2. Moreover, the inserting table 1-2 and the slot 2-6 are made of magnetic materials to achieve vertical suction and fixation of the sample container 1 and the rotating base 2. In the process of rotation, the sample container 1 will be in a relatively stable state to prevent the sample container 1 from swinging. Through the cooperation of the inserting table 1-2 and the slot 2-6, the function of the sample container 1 is guaranteed, and it is easy to disassemble, thereby improving the efficiency of replacing samples.

[0043] One end of the lifting rod 2-2 is fixedly connected to the bottom of the support platform 2-1, and the other end of the lifting rod 2-2 is fixedly connected to the rotating disk 2-3. In this embodiment, the lifting rod 2-2 is an electric lifting rod, which can realize the lifting function and is used to adjust the height of the sample container 1, thereby adaptively adjusting the height of the nutrient solution in the sample container 1.

[0044] Meanwhile, the rotating disk 2-3 is arranged above the base 2-4; the base 2-4 includes a shell 2-5, and the four sides of the shell 2-5 have the same structure; through holes 2-7 are symmetrically opened on both sides of the bottom end of a single side of the shell 2-5, and the two opposite sides of the shell 2-5 are nested on the guide rail of the mobile platform 3 through the through holes 2-7. Specifically, the two through holes 2-7 on the same side of the two opposite sides of the shell 2-5 are nested on the same guide rail.

[0045] During use, a mobile platform 3 with a screw guide rail can be selected to drive the movement of the rotating base through the screw guide rail.

[0046] By nesting the base 2-4 on the guide rail of the mobile platform 3, the rotating base 2 and the sample container 1 can be moved horizontally and vertically on the horizontal plane. At the same time, the lifting rod 2-2 is used to lift the sample container 1, thereby ensuring that the sample container 1 can be moved to any position in the three-dimensional space.

[0047] Meanwhile, the base 2-4 also includes a motor, which is arranged inside the base 2-4, and the central rotating shaft of the motor is fixedly connected to the bottom of the rotating disk 2-3 through the upper surface of the shell 2-5. The motor can drive the rotating disk 2-3 to rotate, thereby driving the sample container 1 to rotate, so that two-dimensional images of cells at different angles can be obtained, which is conducive to the final generation of three-dimensional images of cells.

[0048] Based on the sample container 1 and the rotating base 2 described above, this embodiment proposes a method for using an X-ray-based in-situ visualization device for cell growth, comprising the following steps:

[0049] S1. Place the mobile platform 3 and the detector 6 on the precision translation stage 5, and the rotating base 2 is nested on the guide rail of the mobile platform 3 using the through hole 2-7 on the shell 2-5.

[0050] Among them, the brackets on both sides of the guide rail on the mobile platform 3 are detachable structures, so as to achieve fixed connection with the rotating base 2; and the detector 6 is fixed on the precision translation stage 5 through the corresponding supporting structure, and a certain distance is left between the detector 6 and the upper surface of the precision translation stage 5.

[0051] S2. The nutrient solution containing cells is delivered to the sample container 1 through a syringe, and the nutrient solution is ensured to be stored in the inverted cone-shaped space of the sample container 1 .

[0052] S3. Use the inserting platform 1 - 2 on the side of the sample container 1 to insert and connect the sample container 1 with the slot 2 - 6 of the rotating base 2, and ensure that the sample container 1 is in a vertical state.

[0053] S4. The position of the sample container 1 is adjusted by the lifting rod 2-2 and the moving platform 3 so that the position of the nutrient solution in the sample container 1 and the detector 6 are located on the same horizontal line.

[0054] S5, finally, the nutrient solution in the mobile platform 3 is horizontally irradiated by the X-ray 4 (such as Figure 5 As shown), the rotating disk 2-3 is used to drive the sample container 1 to rotate at a constant speed; the detector 6 receives the X-rays 4 that have irradiated the nutrient solution and cells, and converts them into electrical signals and transmits them to the computer 7. The computer 7 uses phase contrast CT to perform three-dimensional imaging and performs observation and analysis based on the three-dimensional image.

[0055] In order to achieve better imaging, the sample container 1 in this embodiment is made of polyethylene. The polyethylene material itself is transparent and has low X-ray absorption capacity, which will not significantly affect the transmission and scattering of X-rays, thereby helping to generate clearer images and making it easier to observe the fine structure of cells. At the same time, by using X-ray phase contrast CT imaging to perform imaging, there is no need to label cells, thereby avoiding the impact on cell growth and overcoming the shortcomings of fluorescence microscopy imaging that gradually loses fluorescence and affects image clarity and accuracy.

[0056] In addition, due to the radiation dose problem of X-rays, excessive radiation dose will cause heat accumulation, which will lead to radiation loss. The present invention adopts a liquid environment (nutrient solution), which not only provides a growth environment for cells, but also timely removes the heat of the sample during the irradiation process, effectively reducing the risk of radiation damage, which can effectively solve this problem. The phase contrast CT imaging method adopted is an existing technical solution.

[0057] Although the above describes the specific implementation mode of the present invention in conjunction with the accompanying drawings, it is not intended to limit the scope of protection of the present invention. Those skilled in the art should understand that various modifications or variations that can be made by those skilled in the art on the basis of the technical solution of the present invention without creative work are still within the scope of protection of the present invention.

Claims

1. An X-ray based in situ visualization device for cell growth, mounted on a mobile platform with a guide rail, characterized in that: The device comprises a sample container and a rotating base, wherein the rotating base is nested on the guide rail of the mobile platform; the sample container is hollow inside, has no upper bottom surface, and has a cylindrical structure with an inverted cone-shaped space at the bottom. An inserting platform is provided at the bottom of the outer side of the sample container, and the sample container is plug-connected with the slot of the rotating base through the inserting platform.

2. The X-ray based in situ visualization device for cell growth according to claim 1, characterized in that: The sample container is in a vertical state.

3. The X-ray based in situ visualization device for cell growth according to claim 1, characterized in that: The rotating base includes a supporting platform, a lifting rod, a rotating disk and a base, the inserting platform includes a circular positioning boss in the middle and square limiting bosses located at both ends of the positioning boss, the slot includes a circular positioning socket in the middle and square limiting sockets located at both ends of the positioning socket, the positioning boss is inserted into the positioning socket, the limiting boss is inserted into the limiting socket, and the inserting platform and the slot are made of magnetic materials to attract the sample container and the rotating base; one end of the lifting rod is fixedly connected to the bottom of the supporting platform, and the other end of the lifting rod is fixedly connected to the rotating disk, and the rotating disk is arranged above the base.

4. The X-ray based in situ visualization device for cell growth as claimed in claim 3, characterized in that: The lifting rod is specifically an electric lifting rod.

5. The X-ray based in situ visualization device for cell growth as claimed in claim 3, characterized in that: The base includes a shell, and the four sides of the shell have the same structure; through holes are symmetrically opened on both sides of the bottom end of a single side of the shell, and the two opposite sides of the shell are nested on the guide rail of the mobile platform through the through holes.

6. The X-ray based in situ visualization device for cell growth according to claim 5, characterized in that: The base also includes a motor, which is arranged inside the shell. The central rotating shaft of the motor is fixedly connected to the bottom of the rotating disk through the shell.

7. The X-ray based in situ visualization device for cell growth according to claim 1, characterized in that: The inverted cone-shaped space is specifically an inverted circular cone space.

8. The X-ray based in situ visualization device for cell growth according to claim 1, characterized in that: The sample container is made of polyethylene.

9. A method for using an X-ray based in situ visualization device for cell growth according to any one of claims 1 to 8, characterized in that: The specific steps include: S1, placing the mobile platform and the detector on the precision translation stage, and the rotating base is nested on the guide rail of the mobile platform using the through hole on the shell; S2. delivering the nutrient solution containing the cells into the sample container through a syringe, and ensuring that the nutrient solution is stored in the inverted cone space of the sample container; S3, using the insertion platform on the side of the sample container to plug and connect the sample container with the slot of the rotating base, and ensure that the sample container is in a vertical state; S4, adjusting the position of the sample container by means of the lifting rod and the moving platform so that the position of the nutrient solution in the sample container and the detector are on the same horizontal line; S5. Finally, the nutrient solution and cells in the mobile platform are horizontally irradiated with X-rays, and the rotating disk is used to drive the sample container to rotate at a constant speed; the detector receives the X-rays that have irradiated the nutrient solution and cells, and converts them into electrical signals and transmits them to the computer. The computer uses phase contrast CT to perform three-dimensional imaging and performs observation and analysis based on the three-dimensional images.

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